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STUDIES AVAILABLE OF CHITRAKMOOL

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1. Plumbagin, a medicinal plant-derived naphthoquinone, is a novel inhibitor of the growth and invasion of hormone-refractory prostate cancer.

Aziz MH, Dreckschmidt NE, Verma AK.

Department of Human Oncology, School of Medicine and Public Health, University of Wisconsin, Madison, Wisconsin 53792, USA.

Prostate cancer (PCa) is the second leading cause of cancer-related deaths in men. Hormone-refractory invasive PCa is the end stage and accounts for the majority of PCa patient deaths. We present here that plumbagin (PL), a quinoid constituent isolated from the root of the medicinal plant Plumbago zeylanica L., may be a potential novel agent in the control of hormone-refractory PCa. Specific observations are the findings that PL inhibited PCa cell invasion and selectively induced apoptosis in PCa cells but not in immortalized nontumorigenic prostate epithelial RWPE-1 cells. In addition, i.p. administration of PL (2 mg/kg body weight), beginning 3 days after ectopic implantation of hormone-refractory DU145 PCa cells, delayed tumor growth by 3 weeks and reduced both tumor weight and volume by 90%. Discontinuation of PL treatment in PL-treated mice for as long as 4 weeks did not result in progression of tumor growth. PL, at concentrations as low as 5 micromol/L, inhibited in both cultured PCa cells and DU145 xenografts (a) the expression of protein kinase Cepsilon (PKCepsilon), phosphatidylinositol 3-kinase, phosphorylated AKT, phosphorylated Janus-activated kinase-2, and phosphorylated signal transducer and activator of transcription 3 (Stat3); (b) the DNA-binding activity of transcription factors activator protein-1, nuclear factor-kappaB, and Stat3; and (c) Bcl-xL, cdc25A, and cyclooxygenase-2 expression. The results indicate for the first time, using both in vitro and in vivo preclinical models, that PL inhibits the growth and invasion of PCa. PL inhibits multiple molecular targets including PKCepsilon, a predictive biomarker of PCa aggressiveness. PL may be a novel agent for therapy of hormone-refractory PCa.

Publication Types: PMID: 18974148 [PubMed - indexed for MEDLINE]

2. Antimutagenic activity of methanolic extracts of four ayurvedic medicinal plants.

Aqil F, Zahin M, Ahmad I.

Department of Agricultural Microbiology, Aligarh Muslim University, Aligarh 202 002, India.

Methanolic extracts of Acorus calamus (Rhizome), Hemidesmus indicus (Stem), Holarrhena antidysenterica (Bark) and Plumbago zeylanica (Root), were tested for their antimutagenic potential. These extracts, at tested concentrations, showed no sign of mutagenicity to Salmonella typhimurium tester strains. The extracts of the plants exhibited varying level of antimutagenicity. At a dose of 100 microg/plate, the extracts exhibited the inhibition of His+ revertants from 18.51% to 82.66% against direct acting mutagens, methyl methanesulphonate (MMS) and sodium azide (NaN3) induced mutagenicity in Salmonella tester strains TA 97a, TA 100, TA 102 and TA 104. However, at lower concentrations (25 and 50 mcirog/plate) of the plant extracts, a decrease in antimutagenic activity was recorded. Dose dependent antimutagenic activity of the extracts is also evident from linear regression analysis of the data. The over all antimutagenic potential of above four extracts was found to be in order of A. calamus > H. indicus > H. antidysenterica > P. zeylanica. Further, total phenolic content of these extracts did not correlate with its antimutagenic activity in A. calamus and P. zeylanica.

Publication Types:  PMID: 18949897 [PubMed - in process]

3. Seselin from Plumbago zeylanica inhibits phytohemagglutinin (PHA)-stimulated cell proliferation in human peripheral blood mononuclear cells.

Tsai WJ, Chen YC, Wu MH, Lin LC, Chuang KA, Chang SC, Kuo YC.

National Research Institute of Chinese Medicine, No. 155-1, Sec. 2, Li-Nung Street, Shih-Pai, 112, Taipei, Taiwan.

Effects of seselin (C(14)H(12)O(3); MW 228) identified from Plumbago zeylanica on phytohemagglutinin (PHA)-stimulated cell proliferation were studied in human peripheral blood mononuclear cells (PBMC). The data demonstrated that seselin inhibited PBMC proliferation-activated with PHA with an IC(50) of 53.87+/-0.74 microM. Cell viability test indicated that inhibitory effects of seselin on PBMC proliferation were not through direct cytotoxicity. The action mechanisms of seselin may involve the regulation of cell cycle progression, interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) production in PBMC. Since cell cycle analysis indicated that seselin arrested the cell cycle progression of activated PBMC from the G(1) transition to the S phase. Seselin suppressed IL-2 and IFN-gamma production in a concentration-dependent manner. Furthermore, seselin significantly decreased the IL-2 and IFN-gamma gene expression in PHA-activated PBMC. Therefore, results elucidated for the first time that seselin is likely an immunomodulatory agent for PBMC.

Publication Types:  PMID: 18577441 [PubMed - in process]

4. Toxicity studies on dermal application of plant extract of Plumbago zeylanica used in Ethiopian traditional medicine.

Teshome K, Gebre-Mariam T, Asres K, Perry F, Engidawork E.

Department of Pharmacology, School of Pharmacy, Addis Ababa University, Ethiopia.

Plant-based therapeutic preparations are cyclically returning to complement dermatologic therapy, however, data on the toxicity profile of such plants are lacking. In the present study, Plumbago zeylanica, a medicinal plant commonly used in Ethiopia for skin diseases was subjected to a systematic dermatotoxicity study. To this effect, the dermatotoxicity of 80% methanol extract of the root part of Plumbago zeylanica was investigated in animals following standard procedures for irritation, sensitization, acute toxicity and repeated toxicity tests. Extraction of plant material with 80% methanol resulted in 9.45% of crude extract of Plumbago zeylanica. The skin irritation test on rabbits showed Plumbago zeylanica extract to be a moderate irritant, with a primary irritation index of 2.00. Sensitization test on mice by the Mouse Ear Swelling Test method revealed the extract to be non-sensitizer in a dose range of 4-10mg/ml and the percent responder was zero. Acute dermal toxicity test on rats did not produce any overt signs of toxicity, except that there was a weight gain difference between the test and control groups of female rats. This was not, however, supported by other parameters, like the absolute and relative organ weights. Repeated dose toxicity test was associated with increased relative testis weight (P<0.05) as well as higher values for Blood urea nitrogen and K+ (P<0.05) in both sexes with the highest dose (1000 mg/kg) group, although histopathological analyses failed to lend support to these observations. Taken together, the dermatotoxicity test results from this study suggest that Plumbago zeylanica toxic effects might be limited to effects like moderate irritation.

Publication Types:  PMID: 18339496 [PubMed - indexed for MEDLINE]

5. Fungal contamination of raw materials of some herbal drugs and recommendation of Cinnamomum camphora oil as herbal fungitoxicant.

Singh P, Srivastava B, Kumar A, Dubey NK.

Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi, 221005, India.

The paper explores fungal infection and aflatoxin B1 contamination of six medicinal plant samples viz. Adhatoda vasica Nees, Asparagus racemosus Linn., Evolvulus alsinoides Linn., Glycyrrhiza glabra Linn., Plumbago zeylanica Linn. and Terminalia chebula Retz. A total of 858 fungal isolates were detected from the raw materials. Maximum number of fungal isolates was detected from A. racemosus (228). The genus Aspergillus was found to be the most dominant genus causing infection to most of the raw materials. Among the 32 isolates of A. flavus tested, 13 isolates were found to be toxigenic elaborating aflatoxin B1. The highest elaboration of aflatoxin B1 was 394.95 ppb by the isolates of A. flavus from G. glabra. The essential oil of Cinnamomum camphora (L.) Presl showed efficacy in arresting aflatoxin B1 by the toxigenic strain. The growth of a toxigenic strain of A. flavus decreased progressively with increasing concentration of essential oil from leaves of C. camphora. The oil completely inhibited aflatoxin B1 production even at 750 ppm. Hence, the oil of C. camphora is recommended as herbal fungitoxicant against the fungal contamination of the raw materials.

Publication Types:  PMID: 18322727 [PubMed - indexed for MEDLINE]

6. Plumbagin-induced apoptosis in human prostate cancer cells is associated with modulation of cellular redox status and generation of reactive oxygen species.

Powolny AA, Singh SV.

Department of Pharmacology and Urology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.

PURPOSE: To investigate the mechanism of human prostate cancer cell growth inhibition by plumbagin, a constituent of the widely used medicinal herb Plumbago zeylanica L. MATERIALS AND METHODS: Cell viability was determined by trypan blue dye exclusion assay. Apoptosis induction was assessed by analysis of cytoplasmic histone-associated DNA fragmentation. Cell cycle distribution and generation of reactive oxygen species (ROS) were determined by flow cytometry. The effect of plumbagin treatment on cellular redox status was determined by analysis of intracellular glutathione (GSH) levels and expression of genes involved in ROS metabolism. RESULTS: Plumbagin treatment decreased viability of human prostate cancer cells (PC-3, LNCaP, and C4-2) irrespective of their androgen responsiveness or p53 status. Plumbagin-mediated decrease in cell viability correlated with apoptosis induction, which was accompanied by ROS generation and depletion of intracellular GSH levels. Pretreatment of cells with the antioxidant N-acetylcysteine inhibited plumbagin-mediated ROS generation and apoptosis. Plumbagin treatment also resulted in altered expression of genes responsible for ROS metabolism, including superoxide dismutase 2 (Mn-SOD). CONCLUSION: The present study points towards an important role of ROS in plumbagin-induced apoptosis in human prostate cancer cells.

Publication Types: PMID: 18213451 [PubMed - indexed for MEDLINE]

7. Development of an enzyme-linked immunosorbent assay (ELISA) using highly-specific monoclonal antibodies against plumbagin.

Sakamoto S, Putalun W, Tsuchihashi R, Morimoto S, Kinjo J, Tanaka H.

Department of Medicinal Plant Breeding, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.

Plumbagin (PL; 5-hydroxy-2-methyl-1,4-naphthoquinone) is a natural compound mainly isolated from Plumbago zeylanica. This plant is distributed in Southeast Asia, and well known as Ayurvedic medicine in India for its medicinal properties. PL has been shown to have various pharmacological activities. We have successfully prepared monoclonal antibodies against PL, and developed an enzyme-linked immunosorbent assay (ELISA) system for determination of PL. 3-(5-Hydroxy-2-methyl-1,4-naphthoquinone-3-yl) propanoic acid was synthesized and purified to prepare PL-bovine serum albumin conjugate (PL-BSA), which was used as an immunogen. PL-BSA conjugate was administered into BALB/c male mice for production of monoclonal antibodies against PL. The monoclonal antibody against PL which is secreted from established hybridoma cell line 3A3 (MAb 3A3) has been proven to have highly-specific to PL resulting from cross-reactivities test. The range for calibration of PL by ELISA was 0.2-25 microg mL(-1). Based on validation analysis, this analytical method by ELISA is a precise, accurate, and sensitive method for the determination of PL in plant.

Publication Types: PMID: 18155415 [PubMed - indexed for MEDLINE]

8. [Chemical constituents from aerial parts of Plumbago zeylanica Linn]

[Article in Chinese]

Zhang QR, Mei ZN, Yang GZ, Xiao YX

Pharmacy of College, Wuhan University, Wuhan 430072, China.

OBJECTIVE: To investigate the chemical constituents of the aerial parts of Plumbago zeylanica Linn. METHODS: The constituents of the EtOAc-soluble portion in the 95% ethanol extract were isolated and purified by means of chromatography. Compounds were identified by their physical characteristics and spectral features. RESULTS: Nine compounds were isolated as plumbagin (I), isoshinanolone (II), plumbagic acid (III), beta-sitosterol (IV), 4-hydroxybenzaldehyde (V), trans-cinnamic acid (VI), vanillic acid (VII), 2, 5-dimethyl-7-hydroxychromone (VIII), indole-3-carboxaldehyde (IX). CONCLUSION: Compounds V, VII, VIII and IX were isolated for the first time from Plumbago Linn.

Publication Types:  PMID: 17727061 [PubMed - indexed for MEDLINE]

9. Antibacterial properties of traditionally used Indian medicinal plants.

Aqil F, Ahmad I.

Department of Agricultural Microbiology, Aligarh Muslim University, Aligarh, India.

In search of broad-spectrum antibacterial activity from traditionally used Indian medicinal plants, 66 ethanolic plant extracts were screened against nine different bacteria. Of these, 39 extracts demonstrated activity against six or more test bacteria. Twelve extracts showing broad-spectrum activity were tested against specific multidrug-resistant (MDR) bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum beta-lactamases (ESbetaL)-producing enteric bacteria. In vitro efficacy was expressed in terms of minimum inhibitory concentration (MIC) values of plant extracts. MIC values ranged from 0.32-7.5 mg/ml against MRSA and 0.31-6.25 mg/ml against ESbetaL-producing enteric bacteria. The overall activity against all groups of bacteria was found in order of Plumbago zeylanica > Hemidesmus indicus > Acorus calamus > Camellia sinensis > Terminalia chebula > Terminalia bellerica > Holarrhena antidysenterica > Lawsonia inermis > Mangifera indica > Punica granatum > Cichorium intybus and Delonix regia. In addition, these extracts showed synergistic interaction with tetracycline, chloramphenicol and ciprofloxacin against S. aureus and/or Escherichia coli. The ethanolic extracts of more than 12 plants were found nontoxic to sheep erythrocytes and nonmutagenic, determined by Ames test using Salmonella typhimurium test strains (TA 97a, TA 100, TA 102 and TA 104). Based on above properties, six plants-Plumbago zeylanica, Hemidesmus indicus, Acorus calamus, Punica granatum, Holarrhena antidysenterica and Delonix regia-were further subjected to fractionation-based study. Ethyl acetate, acetone and methanol fractions of more than six plants indicated that the active phytocompounds were distributed mainly into acetone and ethyl acetate fractions, whereas they were least prevalent in methanol fractions as evident from their antibacterial activity against MDR bacteria. Gram-positive and Gram-negative MDR bacteria are almost equally sensitive to these extracts/fractions, indicating their broad-spectrum nature. However, strain- and plant extract-dependent variations in the antibacterial activity were also evident. Time-kill assay with the most promising plant fraction Plumbago zeylanica (ethyl acetate fraction) demonstrated killing of test bacteria at the level lower than its MIC. Further, identification of active constituents in each fraction and their additive and synergistic interactions are needed to exploit them in evaluating efficacy and safety in vivo against MDR bacteria. Copyright 2007 Prous Science.

Publication Types: PMID: 17440624 [PubMed - indexed for MEDLINE]

10. [Determination of plumbagin in different parts of Plumbago zeylanica by RP-HPLC]

[Article in Chinese]

Liu Y, Deng F, Liu C, Meng QY, Gao ZW.

Ethnic Pharmaceutical Institute of Southwest University for Nationalities, Chengdu 610041, China.

OBJECTIVE: To develop a RP-HPLC method to determine plumbagin in Plumbago zeylanica, and to investigate contents of plumbagin in different parts of. P. zeylanica. METHOD: The analysis was carried out at 30 degrees C on a Kromasil C18, column eluted with a mobile phase consisting of a mixture of methanol-water (65: 35). The flow rate was 1 mL x min(-1), the detector wavelength was 213 nm. RESULT: The calibration curve was linear within the concentration ranges of 0.020 8-0. 104 microg (r = 0. 9999). The average recovery was 98.7%. The contents in the root, stem and leaf were 0.394 5%, 0.050 8%, 0.031 4% respectively. CONCLUSION: This method is simple, accurate, replicate and suitable for the determination of plumbagin in P. zeylanica.

Publication Types: PMID: 17225534 [PubMed - indexed for MEDLINE]

11. Suberosin inhibits proliferation of human peripheral blood mononuclear cells through the modulation of the transcription factors NF-AT and NF-kappaB.

Chen YC, Tsai WJ, Wu MH, Lin LC, Kuo YC.

Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan, ROC.

BACKGROUND AND PURPOSE: Extracts of Plumbago zeylanica containing suberosin exhibit anti-inflammatory activity. We purified suberosin from such extracts and studied its effects on a set of key regulatory events in the proliferation of human peripheral blood mononuclear cells (PBMC) stimulated by phytohemagglutinin (PHA). EXPERIMENTAL APPROACH: Proliferation of PBMC in culture was measured by uptake of 3H-thymidine; production of cytokines and cyclins by Western blotting and RT-PCR. Transcription factors NF-AT and NF-kappaB were assayed by immunocytochemistry and EMSA. KEY RESULTS: Suberosin suppressed PHA-induced PBMC proliferation and arrested cell cycle progression from the G1 transition to the S phase. Suberosin suppressed, in activated PBMC, transcripts of interleukin-2 (IL-2), interferon-gamma (IFN-gamma), and cyclins D3, E, A, and B. DNA binding activity and nuclear translocation of NF-AT and NF-kappaB induced by PHA were blocked by suberosin. Suberosin decreased the rise in intracellular Ca2+ concentration ([Ca2+]i) in PBMC stimulated with PHA. Suberosin did not affect phosphorylation of p38 and JNK but did reduce activation of ERK in PHA-treated PBMC. Pharmacological inhibitors of NF-kappaB, NF-AT, and ERK decreased expression of mRNA for the cyclins, IL-2, and IFN-gamma and cell proliferation in PBMC activated by PHA. CONCLUSIONS AND IMPLICATIONS: The inhibitory effects of suberosin on PHA-induced PBMC proliferation, were mediated, at least in part, through reduction of [Ca2+]i, ERK, NF-AT, and NF-kappaB activation, and early gene expression in PBMC including cyclins and cytokines, and arrest of cell cycle progression in the cells. Our observations provide an explanation for the anti-inflammatory activity of P. zeylanica.

Publication Types:  PMID: 17179947 [PubMed - indexed for MEDLINE] PMCID: PMC2013892

12. Medicinal plants of the Shinasha, Agew-awi and Amhara peoples in northwest Ethiopia.

Giday M, Teklehaymanot T, Animut A, Mekonnen Y.

Aklilu Lemma Institute of Pathobiology, P.O. Box 1176, Addis Ababa University, Addis Ababa, Ethiopia. mirutseg@yahoo.com

Study was conducted in two sub-districts in northwestern Ethiopia to compile and analyse knowledge on the use of medicinal plants for treatment or prevention of human ailments by three socio-cultural groups, namely the Amharas, Shinashas and Agew-Awis. Data were mainly collected through individual interviews conducted with selected knowledgeable farmers and professional healers of the three socio-cultural groups. A total of 76 medicinal plants belonging to 48 families were documented, of which 50 species were reported by the Amharas, 25 by the Shinashas and 20 by the Agew-Awis. Large proportions of medicinal plants were found to have been used for the treatments of gastro-intestinal complaints (26%), skin diseases (24%) and malaria (22%). Relatively, higher numbers of informants agreed on the use of Croton macrostachyus against malaria (21%), Cynoglossum coeruleum against 'mich', illness mainly characterized by fever, headache and sweating (18%) and Zehneria scabra against malaria (13%). The species Croton Macrostachyus, Calpurnia aurea, Clematis hirsuta and Plumbago zeylanica were found to have the highest diversity of medicinal applications. We recommend that priority for further investigation should be given to medicinal plants with higher informant consensuses, as this could indicate their better efficacy. Measures are needed to conserve plants that are reported as scarce in the study area but still are only harvested from the wild.

Publication Types: PMID: 17101251 [PubMed - indexed for MEDLINE]

13. Evaluation of anti-methicillin-resistant Staphylococcus aureus (MRSA) activity and synergy of some bioactive plant extracts.

Aqil F, Ahmad I, Owais M.

Department of Agricultural Microbiology, Aligarh Muslim University, Aligarh, India. f_aqil@yahoo.com

Anti-methicillin-resistant Staphylococcus aureus (MRSA) activity of ethanolic extracts of four medicinal plants namely Acorus calamus (rhizome) Hemidesmus indicus (stem), Holarrhena antidysenterica (bark), and Plumbago zeylanica (root), were detected with inhibition zone size ranged from 11 to 44 mm and minimum inhibitory concentration (MIC) varied from 0.32 to 3.25 mg/mL. Further, ethyl acetate, acetone and methanol fractions of above plants demonstrated antibacterial activity. The potency of these fractions based on zone of inhibition and MIC value was relatively higher in P. zeylanica (ethylacetate fraction), followed by acetone fractions of H. indicus, A. calamus, and H. antidysenterica. Time kill assay with most promising fractions of these plant extracts, demonstrated concentration-dependent killing of MRSA within 9-12 h of incubation. Interestingly, synergistic interaction among alcoholic extracts and some fractions of above four plants was evident against MRSA. Further, synergistic interaction of these extracts was detected with one or more antibiotics tested (tetracycline, chloramphenicol, ciprofloxacin, cefuroxime and ceftidizime). The findings also validate the traditional uses of above plants against infectious diseases. Phytochemical studies demonstrated flavonoids and phenols as major active constituents. Further investigations are needed to characterize the active principle and its interaction mechanism with antibiotics.

Publication Types:  PMID: 17004300 [PubMed - indexed for MEDLINE]

14. In vitro efficacy of bioactive extracts of 15 medicinal plants against ESbetaL-producing multidrug-resistant enteric bacteria.

Ahmad I, Aqil F.

Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 202002, India. iqbalahmad8@yahoo.co.in

Alcoholic crude extracts and some fractions from 15 traditionally used Indian medicinal plants were investigated for their ability to inhibit the growth of extended spectrum beta-lactamases (ESbetaL)-producing multidrug-resistant enteric bacteria. The test bacteria Eschrichia coli and Shigella were resistant to 16-23 antibiotics with intermediate or resistance to beta-lactams (minimum inhibitory concentration (MIC) value range 16-1024 microg/ml). The crude plant extracts demonstrated zone of inhibition in the range of 11-29 mm against one or more test bacteria. On the basis of promising activity, 12 plants were selected to determine their efficacy in terms of MIC, which ranged from 0.64 mg/ml to 10.24 mg/ml. The extracts of Acorus calamus, Hemidesmus indicus, Holarrhena antidysenterica and Plumbago zeylanica demonstrated relatively high activity as compared to other plant extracts and were fractionated into acetone, ethyl acetate and methanol. Acetone fraction in most of the cases exhibited higher potency (low MIC value) as compared to ethyl acetate and methanol fraction. However, in Plumbago zeylanica, ethyl acetate fraction was most active. Synergistic interactions among crude extracts were demonstrated in the 12 different combinations against ESbetaL-producing E. coli (ESbetaL-02). Certain combinations exhibited significant synergy with enlargement of combined inhibition zone size by 5 mm. Interaction of crude extracts with five antibiotics (Tetracycline, ciprofloxacin, nalidixic acid, chloramphenicol and streptomycin) demonstrated synergistic interaction with tetracycline and ciprofloxacin by 10 and 3 plant extracts respectively. Phytochemical analysis and thin layer chromatography (TLC) bioautography of crude extracts showed the presence of alkaloids, phenols and flavonoids as active phytoconstituents. Most active fractions of four plants were subjected to Infrared spectroscopy and the major groups of compounds were detected. The plant extracts were further tested for their in vitro haemolytic activity to sheep erythrocytes and demonstrated no haemolysis at recommended doses. Further activity-guided fractionation of active fractions is needed to isolate and characterize the active principle in order to establish the mode of action against the ESbetaL-producing multidrug-resistant enteric bacteria and the mechanism of synergy.

Publication Types: PMID: 16875811 [PubMed - indexed for MEDLINE]

15. Measurement and pharmacokinetic study of plumbagin in a conscious freely moving rat using liquid chromatography/tandem mass spectrometry.

Hsieh YJ, Lin LC, Tsai TH.

Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, 155 Li-Nong Street Section 2, Taipei 112, Taiwan.

The aim of the present study is to develop an automated blood sampling (ABS) method coupled to a liquid chromatography-tandem mass spectroscopy (LC-MS/MS) method to evaluate the oral bioavailability of plumbagin in a conscious freely moving rat. Plumbagin, an herbal ingredient, was isolated from Plumbago zeylanica L. The separation was performed using a reversed phase C18 (150mmx4.6mm I.D.; 5microm) column and was eluted with the mobile phase of water-acetonitrile (40:60, v/v) at a flow-rate of 0.8ml/min. Multiple reaction monitoring (MRM) was used to monitor the transition of the deprotonated molecule m/z 187 [MH](-) to the product ion m/z 159 [MHCO](-) for the plumbagin analysis. The calibration curve was linear over the concentration range of 10-2000ng/ml with a coefficient estimation of 0.995. The intra- and inter-day variations (% relative standard deviation; RSD and % bias) of the assay for rat plasma samples were less than 17%. The limit of detection and the limit of quantification were 5 and 10ng/ml, respectively. Recovery of plumbagin from the rat plasma was about 80%. This LC-MS/MS method has been validated to study the pharmacokinetics of plumbagin in rats. The oral bioavailability (AUC(PO)/Dose(PO))/(AUC(IV)/Dose(IV)) of plumbagin was about 38.7+/-5%.

Publication Types: PMID: 16837255 [PubMed - indexed for MEDLINE]

16. Protective effect of Plumbago zeylanica against cyclophosphamide-induced genotoxicity and oxidative stress in Swiss albino mice.

Sivakumar V, Niranjali Devaraj S.

Department of Biochemistry, University of Madras, Chennai, India.

Plumbago zeylanica, commonly known as white leadwort, found abundantly in the plains of Bengal and southern India, was tested for its possible in vivo protective effect against cyclophosphamide-induced genotoxicity and oxidative stress in Swiss albino mice. Pretreatment with the alcoholic root extract of Plumbago zeylanica (250 and 500 mg/kg body weight orally for 5 days) significantly reduced the frequency of micronucleated polychromatic erythrocytes (MnPCEs), increased the PCE/NCE (normochromatic erythrocyte) ratio in the bone marrow, and decreased the levels of lipid peroxidation products with concomitant changes in the status of antioxidants. Both doses of Plumbago zeylanica were effective in exerting a protective effect against cyclophosphamide-induced genotoxicity and oxidative stress.

PMID: 16777706 [PubMed - indexed for MEDLINE]

17. [Effects of plumbagin on the human acute promyelocytic leukemia cells in vitro]

[Article in Chinese]

Zhao YL, Lu DP.

People Hospital, Institute of Hematology, Peking University, Beijing 100044, China.

According to previous clinical experiences of the authors, plumbago zeylanica was effective against acute promyelocytic leukemia (APL). However, its effectiveness has never been proven experimentally or unequivocally clinically. This study was aimed to investigate the effects of plumbagin on the proliferation, cell cycle and apoptosis of APL cell line NB4 Cells. Cell inhibitory rates were detected by MTT colorimetric assay; morphologic changes were observed under light microscope and transmission electron microscope; apoptosis-inducing effects were determined by DNA gel electrophoresis, annexin V/PI double-stained and PI single-stained flow cytometry. The results demonstrated that 2-15 micromol/L plumbagin inhibited the proliferation of NB4 cells in a dose-dependent manner. The morphologic changes of cell apoptosis, such as chromsome condensation and apoptotic body formation, were observed by light microscope and transmission electron microscope. Cell cycle analysis showed that NB4 cells were blocked in G2/M phase of cell cycle. And plumbagin induced annexin V+/PI- cell increase and DNA fragmentation. There was a correlation between cell apoptosis rates and the concentrations of plumbagin in dose-dependent manner (P < 0.05). It is concluded that for the first time the present study shows that plumbagin can inhibit cell proliferation, block cell cycle and induce apoptosis of APL cell line NB4 cells.

Publication Types: PMID: 16638181 [PubMed - indexed for MEDLINE]

18. Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) suppresses NF-kappaB activation and NF-kappaB-regulated gene products through modulation of p65 and IkappaBalpha kinase activation, leading to potentiation of apoptosis induced by cytokine and chemotherapeutic agents.

Sandur SK, Ichikawa H, Sethi G, Ahn KS, Aggarwal BB.

Cytokine Research Laboratory, Department of Experimental Therapeutics, Unit 143, the University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.

Plumbagin, derived from the medicinal plant Plumbago zeylanica, modulates cellular proliferation, carcinogenesis, and radioresistance, all known to be regulated by the activation of the transcription factor NF-kappaB, suggesting plumbagin might affect the NF-kappaB activation pathway. We found that plumbagin inhibited NF-kappaB activation induced by TNF, and other carcinogens and inflammatory stimuli (e.g. phorbol 12-myristate 13-acetate, H2O2, cigarette smoke condensate, interleukin-1beta, lipopolysaccharide, and okadaic acid). Plumbagin also suppressed the constitutive NF-kappaB activation in certain tumor cells. The suppression of NF-kappaB activation correlated with sequential inhibition of the tumor necrosis factor (TNF)-induced activation of IkappaBalpha kinase, IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 phosphorylation, p65 nuclear translocation, and the NF-kappaB-dependent reporter gene expression activated by TNF, TNFR1, TRAF2, NIK, IKK-beta, and the p65 subunit of NF-kappaB. Plumbagin also suppressed the direct binding of nuclear p65 and recombinant p65 to the DNA, and this binding was reversed by dithiothreitol both in vitro and in vivo. However, plumbagin did not inhibit p65 binding to DNA when cells were transfected with the p65 plasmid containing cysteine 38 mutated to serine. Plumbagin down-regulated the expression of NF-kappaB-regulated anti-apoptotic (IAP1, IAP2, Bcl-2, Bcl-xL, cFLIP, Bfl-1/A1, and survivin), proliferative (cyclin D1 and COX-2), and angiogenic (matrix metalloproteinase-9 and vascular endothelial growth factor) gene products. This led to potentiation of apoptosis induced by TNF and paclitaxel and inhibited cell invasion. Overall, our results indicate that plumbagin is a potent inhibitor of the NF-kappaB activation pathway that leads to suppression of NF-kappaB-regulated gene products. This may explain its cell growth modulatory, anticarcinogenic, and radiosensitizing effects previously described.

Publication Types :  PMID: 16624823 [PubMed - indexed for MEDLINE]

   

19. Plumbago zeylanica action on blood coagulation profile with and without blood volume reduction.

Vijayakumar R, Senthilvelan M, Ravindran R, Devi RS.

Department of Physiology, Dr. ALM PG Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai-113, India.

Plumbago zeylanica (PZ) is extensively used in Indian systems of medicine for its medicinal properties. The structure of its active principle is similar to that of vitamin K. Its effect on blood coagulation profile after chronic administration has not been reported so far. The PZ extract (2 mg/kg body weight) and napthoquinone (2 mg/kg body weight) given to individual groups were screened for its effect on bleeding time (BT), clotting time (CT), prothrombin time (PT), platelet count and platelet adhesion in albino rats after 1-day, 15-day and 31-day treatment. There was no change in the platelet count in the treated groups when compared to the control levels. But the platelet adhesion was significantly decreased after PZ and also napthaquinone-treated animals in both with and without blood volume reduction after 15th as well as 31st day. Since the napthoquinone-treated group also showed similar response the changes observed after PZ treatment may be due to this component. Even at a lower dosage level (2 mg/kg body weight), the chronic PZ administration prolongs the bleeding time by altering platelet adhesiveness and the coagulation.

Publication Types:  PMID: 16531123 [PubMed - indexed for MEDLINE]

20. A highly efficient in vitro plant regeneration system and Agrobacterium-mediated transformation in Plumbago zeylanica.

Wei X, Gou X, Yuan T, Russell SD.

Department of Botany and Microbiology, University of Oklahoma, Norman, OK 73019, USA.

Plumbago zeylanica is a unique model for studying flowering plant gametogenesis, heterospermy, and preferential fertilization, yet understanding the control of related molecular mechanisms is impossible without efficient and reproducible regeneration and stable genetic transformation. We found three key factors for enhancing successful regeneration: (1) tissue source of explants, (2) combination and concentration of growth regulators, and (3) culture conditions. The highest frequency of shoot regeneration was achieved using hypocotyl segments cultured on MS basal medium supplemented with BA 2.0 mg/l, NAA 0.75 mg/l, adenine 50 mg/l and 10% (v/v) coconut milk under subdued light at 25+/-2 degrees C; under these conditions, each hypocotyl segment produced over 30 shoots, arising primarily through direct organogenesis after 3 weeks of culture. Regenerated shoots rooted easily on half-strength basal MS medium and were successfully established in the greenhouse. Using this tissue culture protocol, reporter gene GUS under the constitutive CaMV 35S promoter was introduced into P. zeylanica cells of petiole, cotyledon and hypocotyl with A. tumefaciens strains AGL1 and LBA4404. Transient expression was observed in all recipient tissues. Stable transgenic calli originating from petiole were obtained.

Publication Types: PMID: 16470412 [PubMed - indexed for MEDLINE]

21. Antioxidant activity of a salt-spice-herbal mixture against free radical induction.

Natarajan KS, Narasimhan M, Shanmugasundaram KR, Shanmugasundaram ER.

ALMPG Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai 600113, India.

A combination of spices (Piper nigrum, Piper longum and Zingiber officinale), herbs (Cyperus rotundus and Plumbago zeylanica) and salts make up Amrita Bindu. The study was focused to evaluate the antioxidant property of individual ingredients in Amrita Bindu against the free radical 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS). The analysis revealed the antioxidant potential of the ingredients in the following order: Piper nigrum>Piper longum>Cyperus rotundus>Plumbago zeylanca>Zingiber officinale. Two different experiments were designed. In experiment I, rats were fed with normal diet whereas in experiment II rats were given feed mixed with Amrita Bindu for 3 weeks (4 g/kg of feed). Rats from both experimental groups were challenged against a single intraperitonial injection of phenylhydrazine (PHZ) (7.5 mg/kg body weight). At the end of 24 and 72 h, blood was analysed for free radicals and antioxidant levels. It was interesting to note that rats with Amrita Bindu pretreatment showed significantly lower levels of free radicals, lipid peroxidation and protein carbonyls along with significantly higher levels of antioxidants when compared with rats without Amrita Bindu pretreatment on PHZ administration. These results reveal that Amrita Bindu, a salt-spice-herbal mixture exerts a promising antioxidant potential against free radical induced oxidative damage.

PMID: 16337350 [PubMed - indexed for MEDLINE]

22. In vivo micronucleus assay and GST activity in assessing genotoxicity of plumbagin in Swiss albino mice.

SivaKumar V, Prakash R, Murali MR, Devaraj H, Niranjali Devaraj S.

Department of Biochemistry, University of Madras, Guindy Campus, Chennai, India.

Information available on the mutagenicity of a large number of indigenous drugs commonly employed in the Siddha and Ayurveda systems of medicine is scanty. In this context, the current investigation on plumbagin, 5-hydroxy-2methyl-1,4-napthoquinone, an active principle in the roots of Plumbago zeylanica used in Siddha and Ayurveda for various ailments, was carried out; 16 mg/kg b.w. (LD(50)) was fixed as the maximum dose. Subsequent dose levels were fixed as 50% and 25% of LD(50) amounting to 8 mg and 4 mg/kg b.w., respectively, and given orally for 5 consecutive days in 1% Carboxyl Methyl Cellulose (CMC) to Swiss albino mice weighing 25-30 g. The micronucleus assay was done in mouse bone marrow. Plumbagin was found to induce micronuclei at all the doses studied (4 mg/kg, 8 mg/kg, 16 mg/kg b.w.), and it proves to be toxic to bone marrow cells of Swiss albino mice. Animal treated with cyclophosphamide (40 mg/kg b.w.) served as positive control. In addition, glutathione S-transferase (GST) activity was observed in control, plumbagin (4 mg, 8 mg, 16 mg/kg b.w., respectively), and genotoxin-treated experimental group of animals. No significant change in GST activity was observed with plumbagin dose of 4 mg/kg b.w., whereas GST activity was significantly inhibited by higher doses of plumbagin (8 mg and 16 mg/kg b.w.) and cyclophosphamide.

PMID: 16298878 [PubMed - indexed for MEDLINE]

23. High-performance liquid chromatography for quantification of plumbagin, an anti-Helicobacter pylori compound of Plumbago zeylanica L.

Wang YC, Huang TL.

Department of Food Science and Biotechnology, National Chung Hsing University, Taichung 402, Taiwan. ycwang@dragon.nchu.edu.tw

The plant Plumbago zeylanica L. is a semi-climbing shrub that grows throughout Asia and Africa. In our previous study, P. zeylanica L. exhibited high anti-Helicobacter pylori and good bactericidal activities over a wide pH range (pH 2-7). Plumbagin - the major ingredient derived from the roots of P. zeylanica L. - is a naphthoquinone compound. In this study, we investigated plumbagin's anti-H. pylori activity and developed a reversed-phase high-performance liquid chromatography (HPLC) method for quantification of plumbagin from P. zeylanica L. We also observed that plumbagin has strong anti-H. pylori activity, with 0.02-0.16 mg/ml as minimum inhibitory concentrations and 0.16-1.28 mg/ml as minimum bactericidal concentrations. Reversed-phase HPLC was performed with a gradient mobile phase composed of water and methanol, and peaks were detected at 254 nm. Standard curves were linearized in the range of from 10 to 200 microg/ml (regression coefficient r2 = 0.99995). After spikes of 50, 100, and 150 microg/ml of plumbagin standard solution, recovery rates were between 97.45 and 99.24%. Both intra- and inter-day precisions had coefficient variation of less than 1% at concentrations of 50, 100, and 150 microg/ml. The limits of detection and quantitation were 0.02 and 0.06 microg/ml, respectively. Based on validation results, this analytical method is a precise, accurate and stable method to quantify plumbagin derived from P. zeylanica L.

Publication Types: PMID: 16257295 [PubMed - indexed for MEDLINE]

24. Antiviral activities of some Ethiopian medicinal plants used for the treatment of dermatological disorders.

Gebre-Mariam T, Neubert R, Schmidt PC, Wutzler P, Schmidtke M.

Department of Pharmaceutics, School of Pharmacy, Addis Ababa University, P.O. Box, 1176 Addis Ababa, Ethiopia.

Acokanthera schimperi (Apocynaceae), Euclea schimperi (Ebenaceae), Inula confertiflora (Asteraceae), Melilotus elegans (Leguminosae), and Plumbago zeylanica (Plumbaginaceae), are some of the medicinal plants used in Ethiopia for treatment of various skin disorders. In this study, the antiviral activities of the 80% methanolic extracts of these plants have been examined against coxsackievirus B3 (CVB3), influenza A virus and herpes simplex virus type1 Kupka (HSV-1) using cytopathic effect (CPE) inhibitory assays in HeLa, MDCK, and GMK cells, respectively. In parallel, the cytotoxicity was quantified using a crystal violet uptake assay. The antiviral activity of the most active compound was confirmed with plaque reduction assays. The results revealed that the extracts of Acokanthera schimperi and Euclea schimperi showed antiviral activity against all three tested viruses albeit with unequal efficacy. Whereas the Acokanthera schimperi extract exhibited the strongest activity against CVB3, the extract of Euclea schimperi inhibited influenzavirus A replication most effectively. A weak anti-influenzavirus A activity was also exhibited by the other plant extracts tested. In addition, CVB3 was inhibited by the extracts of Plumbago zeylanica and HSV-1 by Inula confertiflora. Thus, the extracts of these plants, particularly those of Acokanthera schimperi, Euclea schimperi and Inula confertiflora which showed activity against CVB3 and HSV-1 support their traditional use in the treatment of skin diseases of viral origin.

Publication Types: PMID: 16233967 [PubMed - indexed for MEDLINE]

25. Determination and identification of plumbagin from the roots of Plumbago zeylanica L. by liquid chromatography with tandem mass spectrometry.

Hsieh YJ, Lin LC, Tsai TH.

Institute of Traditional Medicine, National Yang-Ming University, Taipei, Taiwan.

Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) is an herbal ingredient which is isolated from the root of Plumbago zeylanica L. This herb is a semi-climbing subshrub distributed in thickets or grassland at low elevations of Taiwan. The crushed roots of P. zeylanica L. were ground from lumps to powder and boiled with H2O, 50% EtOH, or 95% EtOH. Chromatographic separation of plumbagin from the herb was carried out using a ZORBAX Extend-C18 column (150 x 4.6 mm I.D.; 5 microm) that was eluted with the mobile phase of water-methanol (10:90, v/v). Multiple reaction monitoring (MRM) was used to monitor the transition of the deprotonated molecule m/z 187 [M-H]- to the product ion m/z 159 [M-H-CO]- for plumbagin analysis. The limit of quantification was determined to and accuracy of 1 ng/ml. Furthermore, the mass fractions of plumbagin in P. zeylanica L. for H2O, 50% EtOH and 95% EtOH were 0.24 +/- 0.04, 3.92 +/- 0.87 and 13.4 +/- 1.59 g/kg, respectively. These results present a reliable liquid chromatography coupled with tandem mass spectrometric (LC-MS/MS) method for the determination of plumbagin form herbal medicines.

Publication Types: PMID: 16078700 [PubMed - indexed for MEDLINE]

26. Anti-Helicobacter pylori activity of Plumbago zeylanica L.

Wang YC, Huang TL.

Department of Food Science, National Chung-Hsing University, 250 Kuokuang Road, Taichung 40227, Taiwan, 402, ROC. ycwang@dragon.nchu.edu.tw

It has been shown that the presence of infection by Helicobacter pylori is strongly associated with gastric cancer and peptic ulceration. In western medicine, a 3-fold therapeutic regimen, emphasizing the use of antibiotics, is typically used to suppress H. pylori activity. However, antibiotic drug resistance frequently develops as a consequence of such treatment. In our previous study, 50 Taiwanese folk medicinal plants were screened for their anti-H. pylori activities. The results revealed that Plumbago zeylanica L. had the highest inhibitory effects against H. pylori. In this study, therefore, we have focused on establishing the anti-H. pylori activities of P. zeylanica L. Water and the organic solvents ethanol, ethyl acetate and acetone were used for P. zeylanica L. extraction, obtaining yields of 1.66-6.84% (w/w). Excluding the water extract, higher anti-H. pylori activity was demonstrated for all the extracts, both using the agar diffusion and dilution methods. The ethyl acetate extract exhibited the lowest minimum inhibitory concentrations against five H. pylori strains, of which ranged from 0.32 to 1.28 mg ml-1, followed, in ascending order, by the acetone, ethanol and water analogs. Bactericidal activity was determined for P. zeylanica L. extracts, with the lowest minimum bactericidal concentrations (5.12-20.48 mg ml-1) demonstrated for the ethyl acetate, followed, in ascending order, by the acetone and ethanol analogs. Bactericidal activity appeared to be in a dose-dependent manner. Through a broad pH range (2-7), bactericidal activity was not affected when extract concentrations were greater than or equal to the minimum bactericidal concentration. High stability was demonstrated for the ethyl acetate P. zeylanica L. extract within pH range of 1-7, exhibiting all pH treatments bactericidal activity.

PMID: 15708315 [PubMed - indexed for MEDLINE]

27. Screening of anti-Helicobacter pylori herbs deriving from Taiwanese folk medicinal plants.

Wang YC, Huang TL.

Department of Food Science, National Chung-Hsing University, 250 Kuokuang Road, Taichung 40227, Taiwan, 402, ROC. ycwang@dragon.nchu.edu.tw

In this study, extracts from 50 Taiwanese folk medicinal plants were examined and screened for anti-Helicobacter pylori activity. Ninety-five percent ethanol was used for herbal extraction. Paederia scandens (Lour.) Merr. (PSM), Plumbago zeylanica L. (PZL), Anisomeles indica (L.) O. Kuntze (AIOK), Bombax malabaricum DC. (BMDC) and Alpinia speciosa (J. C. Wendl.) K. Schum. (ASKS) and Bombax malabaricum DC. (BMDC) all demonstrated strong anti-H. pylori activities. The minimum inhibitory concentration values of the anti-H. pylori activity given by the five ethanol herb extracts ranged from 0.64 to 10.24 mg ml(-1). Twenty-six herbs, including Artemisia argvi Levl. et Vant (AALEV), Phyla nodiflora (Linn.) Greene (PNG) and others, showed moderate anti-H. pylori activity. The additional 19 herbs, including Areca catechu Linn. (ACL), Euphorbia hirta Linn. (EHL) and Gnaphalium adnatum Wall. ex DC. (GAWEDC), possessed lower anti-H. pylori effects. About half of the Taiwanese folk medicinal plants tested, demonstrated to possess higher anti-H. pylori activity.

PMID: 15681161 [PubMed - indexed for MEDLINE]

28. Antimycobacterial constituents from Juniperus procera, Ferula communis and Plumbago zeylanica and their in vitro synergistic activity with isonicotinic acid hydrazide.

Mossa JS, El-Feraly FS, Muhammad I.

Department of Pharmacognosy, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.

The synergistic activity of antimycobacterial constituents from Saudi plants was evaluated in combination with isonicotinic acid hydrazide (INH) against four atypical organisms, namely, Mycobacterium intracellulare, M. smegmatis, M. xenopei and M. chelonei. The potency of INH was increased four-fold, using an in vitro checkerboard method, against each mycobacteria when tested with a subtoxic concentration of the totarol, isolated from J. procera. The MIC values of totarol, ferulenol (from Ferula communis) and plumbagin (from Plumbago zeylanica) were thus lowered from 1.25-2.5 to 0.15-0.3 microg/mL due to synergism with INH. When tested against the resistant strain of M. tuberculosis H37Rv, plumbagin and 7beta-hydroxyabieta-8,13-dien-11,12-dione exhibited inhibitory activity at <12.5 microg/mL, while others were inactive at this concentration. Copyright 2004 John Wiley & Sons, Ltd.

PMID: 15597311 [PubMed - indexed for MEDLINE]

29. Antioxidant properties of Plumbago zeylanica, an Indian medicinal plant and its active ingredient, plumbagin.

Tilak JC, Adhikari S, Devasagayam TP.

Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Mumbai, India.

Plumbago zeylanica (known as "Chitrak") is a useful Indian medicinal plant. The root of the plant and its constituents are credited with potential therapeutic properties including anti-atherogenic, cardiotonic, hepatoprotective and neuroprotective properties. To examine possible mechanisms of action of P. zeylanica (Chitrak), in relation to its reported beneficial properties, antioxidant effects of the aqueous/alcoholic extracts of root, corresponding to medicinal preparations, and the active ingredient, plumbagin, were studied. Methods used included: ferric reducing/antioxidant power (FRAP), radical scavenging of 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2'-azobis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS), lipid peroxidation in rat liver mitochondria induced by different agents, and estimating phenolic and flavonoid content. In FRAP/DPPH assays, boiled ethanolic extracts were the most effective, while in the ABTS assay boiled aqueous extracts were the most efficient. These extracts also significantly inhibited lipid peroxidation induced by cumene hydroperoxide, ascorbate-Fe(2+) and peroxynitrite and contained high amounts of polyphenols and flavonoids. To examine the mechanisms of action in detail, antioxidant and pulse radiolysis studies with plumbagin were conducted. The hydroxyl (.OH), alkyl peroxyl (CCl(3)OO.), linoleic acid peroxyl (LOO.), and glutathiyl (GS.) radicals generate a phenoxyl radical upon reaction with plumbagin. The bimolecular rate constants were: .OH, 2.03 x 10(9) dm(3)mol(-1)s(-1); CCl(3)OO., 1.1 x 10(9) dm(3)mol(-1)s(-1); LOO., 6.7 x 10(7) dm(3)mol(-1)s(-1); and GS., 8.8 x 10(8) dm(3)mol(-1)s(-1). In conclusion, our studies reveal that extracts of P. zeylanica and its active ingredient plumbagin have significant antioxidant abilities that may possibly explain some of the reported therapeutic effects.

PMID: 15479566 [PubMed - indexed for MEDLINE]

30. Cytotoxic constituents from Plumbago zeylanica.

Nguyen AT, Malonne H, Duez P, Vanhaelen-Fastre R, Vanhaelen M, Fontaine J.

Laboratoire de Physiologie et de Pharmacologie Fondamentales, Institut de Pharmacie CP-205/7, Université Libre de Bruxelles, B-1050 Brussels, Belgium. annhthng@ulb.ac.be

The bioassay-guided fractionation of the dichloromethane extract of aerial parts of Plumbago zeylanica led to the isolation of beta-sitosterol, beta-sitosteryl-3beta-glucopyranoside, beta-sitosteryl-3beta-glucopyranoside-6'-O-palmitate (1), lupenone, lupeol acetate, plumbagin and trilinolein. Compound 1 showed cytotoxic activity against MCF7 and Bowes cancer cell lines (IC50 113 microM and 152 microM, respectively), beta-sitosterol inhibited Bowes cell growth (IC50 36.5 microM) and plumbagin was cytotoxic against MCF7 and Bowes cells (IC50 1.28 microM and 1.39 microM, respectively).

Publication Types: PMID: 15261389 [PubMed - indexed for MEDLINE]

31. Inhibition of immediate allergic reactions by ethanol extract from Plumbago zeylanica stems.

Dai Y, Hou LF, Chan YP, Cheng L, But PP.

Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, China. yuedaicpu@hotmail.com

The antiallergic properties of the 70% ethanol extract from Plumbago zeylanica stems (EPZ) were investigated in the present study. The extract (500, 1000 mg/kg, p.o.) dose-dependently inhibited systemic anaphylactic shock induced by compound 48/80 in mice, reduced homologous passive cutaneous anaphylaxis and skin reactions induced by histamine or serotonin in rats, significant differences were observed at the dose of 1000 mg/kg. In vitro, EPZ (5, 20, 50 microg/ml) concentration-dependently reduced histamine release from rat peritoneal mast cells caused by compound 48/80 and antigen. EPZ (50 microg/ml) markedly increased intracellular cAMP content of rat mast cells. These findings demonstrate that EPZ inhibits mast cell-dependent immediate allergic reactions, which is probably mediated by reducing the release of mediators such as histamine from mast cells via elevating intracellular cAMP level and weakening the inflammatory action of mediators.

Publication Types :PMID: 14993817 [PubMed - indexed for MEDLINE]

32. [Effects of the water-soluble extracts from the single herb of ganduqing against hepatitis B virus in vitro]

[Article in Chinese]

Chen W, Yu Z, Li S.

Research Institute of Liver Diseases, Guangzhou Air Force Hospital, 510602.

Comparing with Ara-Amp, the effects of the water-soluble extracts from the single herb of the formula for Ganduqing on HBeAg and HBsAg expression in 2.2.15 cells were studied. The results showed that the extracts of Serissa serissoides (DC) Druce, Hibiseus mutabilis Linn, Paedeuia scangens (Lour) Merr var tomentosa (BL) Hand-Mazz, Plumbago zeylanica L, Garcinia oblougifolia Champ and Begpnia edulia Levl had marked inhibition effects on HBeAg and HBsAg which expressed by 2.2.15 cells.

Publication Types:  PMID: 12571922 [PubMed - indexed for MEDLINE]

33. Cytotoxic naphthoquinones and plumbagic acid glucosides from Plumbago zeylanica.

Lin LC, Yang LL, Chou CJ.

National Research Institute of Chinese Medicine, Pettou, Taipei 112, Taiwan, ROC. lclin@cma23.nricm.edu.tw

Two plumbagic acid glucosides, 3'-O-beta-glucopyranosyl plumbagic acid and 3'-O-beta-glucopyranosyl plumbagic acid methylester along with five naphthoquinones (plumbagin, chitranone, maritinone, elliptinone and isoshinanolone), and five coumarins (seselin, 5-methoxyseselin, suberosin, xanthyletin and xanthoxyletin) were isolated from the roots of Plumbago zeylanica. All coumarins were not previously found in this plant. Cytotoxicity of these compounds to various tumor cells lines was evaluated, and plumbagin significantly suppressed growth of Raji, Calu-1, HeLa, and Wish tumor cell lines.

Publication Types:  PMID: 12560036 [PubMed - indexed for MEDLINE]

34. Central nervous system stimulatory action from the root extract of Plumbago zeylanica in rats.

Bopaiah CP, Pradhan N.

Department of Psychopharmacology, National Institute of Mental Health and Neurosciences, Bangalore - 560029, India.

The effects of a 50% ethanol extract of the root of Plumbago zeylanica (P. zeylanica) were investigated on locomotor behaviour and central dopaminergic activity in rats. The effects on the ambulatory behaviour were assessed along with the levels of dopamine (DA) and its metabolite homovanillic acid (HVA) in the striatum after a single oral dose (100, 200 and 300 mg/kg body weight) of the extract. The extract significantly increased the spontaneous motility in animals. The ambulatory and rotatory behaviour in the treated groups were higher than in the control group (p < 0.05). There were marked differences in the ambulatory behaviour between 100 and 300 mg/kg, indicating that the responses were stimulatory and dose-dependent. The stereotypic behaviour which is characteristic of a dopamine agonist showed biphasic effects. However, there was no significant difference between the groups (p > 0.05). The results showed that the extract of the root of P. zeylanica specifically enhanced the spontaneous ambulatory activity without inducing stereotypic behaviour. The neurochemical estimations revealed elevated levels of DA and HVA in striatum compared with the control rats (p < 0.01). The levels were higher for the 100 mg/kg treated group than the other groups. The levels declined by increasing the dosage of the extract to 200 mg/kg and 300 mg/kg, however, these levels remained higher than the control group. The relationship between motor activity and levels of dopamine are not parallel. These behavioural and biochemical results indicated stimulatory properties of the extract of the root of P. zeylanica, which may be mediated by dopaminergic mechanisms in the rat brain. Copyright 2001 John Wiley & Sons, Ltd.

PMID: 11268117 [PubMed - indexed for MEDLINE]

35. In vitro screening of Indian medicinal plants for antiplasmodial activity.

Simonsen HT, Nordskjold JB, Smitt UW, Nyman U, Palpu P, Joshi P, Varughese G.

Department of Medicinal Chemistry, Royal Danish School of Pharmacy, Group of Pharmacognosy, Universitetsparken 2, 2100 Copenhagen, Denmark.

Plants traditionally used in India to treat fever or malaria were examined in vitro for antiplasmodial properties against Plasmodium falciparum. Of 80 analysed ethanol extracts, from 47 species, significant effects were found for 31 of the extracts. These represent 23 different species from 20 families. Of the active species 20 were tested against P. falciparum for the first time. The following five species seems to be of special interest for further antimalarial studies, Casearia elliptica, Holarrhena pubescens, Pongamia pinnata, Soymida febrifuga, and Plumbago zeylanica.

Publication Types: PMID: 11167038 [PubMed - indexed for MEDLINE]

36. An investigation into the biochemical basis of the observed hyperglycaemia in rats treated with ethanol root extract of plumbago zeylanica.

Olagunju JA, Jobi AA, Oyedapo OO.

Department of Biochemistry, Obafemi Awolowo College of Health Science, Ogun State University, Ago-Iwoye, Nigeria.

The effects of the ethanol extract of the root of Plumbago zeylanica on key enzymes of glycolysis and other biochemical parameters were studied in the rat. The results show that thigh muscle hexokinase, phosphofructokinase, pyruvate kinase and lactate dehydrogenase activities were significantly reduced (p < 0.05) by 12.07%, 51.02%, 24.32% and 25.16% respectively in rats treated with the ethanol extract of Plumbago zeylanica when compared with the controls. Serum pyruvate and lactate were significantly lowered in the experimental rats by 23.64% and 46.29%, respectively. The difference between the supernatant protein means was not statistically different (p > 0.05) suggesting the preservation of protein synthesis in the muscle of the extract-treated rats. The reduction in the activities of the key enzymes of glycolysis and its end-products suggests a reduction in flux across the glycolytic pathway in the extract-treated rats. This may be a result of impaired delivery to, and utilization of, glucose by the peripheral tissue, thus substantiating the reported hyperglycaemia in the extract-treated rats.

PMID: 10404546 [PubMed - indexed for MEDLINE]

37. Isolation and collection of two populations of viable sperm cells from the pollen of Plumbago zeylanica.

Zhang Z, Xu H, Singh MB, Russell SD.

Department of Botany and Microbiology, University of Oklahoma, Norman 73019-0245, USA.

A protocol is described for individually collecting two populations of sperm cells, Svn and Sua, from pollen of Plumbago zeylanica. Pollen grains were burst in 10 mM MOPS buffer containing 0.8 M mannitol (pH 4.6). Paired sperm cells released from pollen were separated using a microinjector. Svn and Sua were then collected individually with a microinjector, based upon known size differences. Collected sperm cells were washed with isolation medium and transferred to liquid nitrogen until use. Fluorochromatic reaction (FCR) test of isolated sperm cells showed a positive reaction, indicating that the isolated sperm cells are viable; most of the sperm cells retain viability for at least 2 h.

Publication Types: PMID: 9921639 [PubMed - indexed for MEDLINE]

38. Screening of some Indian medicinal plants for their antimicrobial properties.

Ahmad I, Mehmood Z, Mohammad F.

Department of Agricultural Microbiology, Institute of Agriculture, Aligarh Muslim University, India.

A total of 82 Indian medicinal plants traditionally used in medicines were subjected to preliminary antibacterial screening against several pathogenic and opportunistic microorganisms. Aqueous, hexane and alcoholic extracts of each plant were tested for their antibacterial activity using agar well diffusion method at sample concentration of 200 mg/ml. The results indicated that out of 82 plants, 56 exhibited antibacterial activity against one or more test pathogens. Interestingly, extracts of five plants showed strong and broad spectrum activity as compared to rest of 51 plant extracts which demonstrated moderate activity. On the whole the alcoholic extracts showed greater activity than their corresponding aqueous and hexane extracts. Among various extracts, only alcoholic extracts of Emblica officinalis, Terminalia chebula, Terminalia belerica, Plumbago zeylanica and Holarrhena antidysenterica were found to show potentially interesting activity against test bacteria. These active crude alcoholic extracts were also assayed for cellular toxicity to fresh sheep erythrocytes and found to have no cellular toxicity.

PMID: 9741890 [PubMed - indexed for MEDLINE]

39. Modulatory effect of plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) on macrophage functions in BALB/c mice. I. Potentiation of macrophage bactericidal activity.

Abdul KM, Ramchender RP.

Department of Zoology, Osmania University, Hyderabad, India.

The modulatory ability of plumbagin, a natural product from Plumbago zeylanica, was studied on peritoneal macrophages of BALB/c mice. The macrophage functions evaluated were bactericidal activity, hydrogen peroxide and superoxide anion release. The bactericidal capacity of in vivo plumbagin-treated mouse macrophages was estimated against Staphylococcus aureus. In low doses plumbagin exerted a constant increase in bactericidal activity throughout the study period whereas with a high dose a higher response was observed up to six weeks. But in the next two weeks a considerable decline in the bactericidal activity was noticed compared to low dose. Plumbagin was also seen to exert a similar response on oxygen radical release by macrophages in vivo showing a clear correlation between oxygen radical release and the bactericidal activity. The data indicate that plumbagin augments the macrophage bactericidal activity by potentiating the oxyradical release at low concentration whereas at the higher concentration it has inhibitory activity.

Publication Types:  PMID: 8557523 [PubMed - indexed for MEDLINE]

40. Effect of Plumbagin on some glucose metabolising enzymes studied in rats in experimental hepatoma.

Parimala R, Sachdanandam P.

Department of Medical Biochemistry, University of Madras, India.

Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) isolated from Plumbago zeylanica Linn, when administered orally, at a dosage of 4 mg/kg body weight induces tumour regression in 3-methyl-4-dimethyl aminoazobenzene (3MeDAB) induced hepatoma in Wistar male rats. The purpose of this investigation was to identify the changes in the rate of glycolysis and gluconeogenesis in tumour-bearing rats and the effects of treatment with Plumbagin. The levels of certain glycolytic enzymes, namely, hexokinase; phosphoglucoisomerase; and aldolase levels increased (p < 0.001) in hepatoma bearing rats, whereas they decreased in Plumbagin administered rats to near normal levels. Certain gluconeogenic enzymes, namely, glucose-6-phosphatase and fructose-1,6-diphosphatase decreased (p < 0.001) in tumour hosts, whereas Plumbagin administration increased the gluconeogenic enzyme levels in the treated animals. These investigations indicate the molecular basis of the different biological behaviour of 3MeDAB induced hepatoma and the anticarcinogenic property of Plumbagin against hepatoma studied in rats.

Publication Types: PMID: 8264573 [PubMed - indexed for MEDLINE]

41. Cytoskeletal organisation and modification during pollen tube arrival, gamete delivery and fertilisation in Plumbago zeylanica.

Huang BQ, Pierson ES, Russell SD, Tiezzi A, Cresti M.

Department of Botany and Microbiology, University of Oklahoma, Norman 73019-0245.

The cytoskeletal organisation of the isolated embryo sac and egg cells of Plumbago zeylanica was examined before, during and after pollen tube penetration into the embryo sac to determine the potential involvement of microtubules and actin filaments in fertilisation. Material was singly and triply stained using Hoechst 33258 to localise DNA, fluorescein isothiocyanate (FITC)-labelled anti-alpha-tubulin to detect microtubules and rhodamine-phalloidin to visualise F-actin. Microtubules in the unfertilised egg cell are longitudinally aligned in the micropylar and mid-lateral areas, aggregating into bundles near the filiform apparatus. In the perinuclear cytoplasm of the egg cell, microtubules become more or less randomly aligned. F-actin bundles form a longitudinally aligned mesh in the chalazal cytoplasm of the egg cell. In the central cell, microtubules and F-actin are distributed along transvacuolar strands and are also evident in the perinuclear region and at the periphery of the cell. During pollen tube penetration, sparse microtubule bundles near the pathway of the pollen tube may form an apparent microtubular 'conduit' surrounding the male gametes at the delivery site. Actin aggregates become organised near the pathway of the pollen tube and at the delivery site of the sperm cells. Subsequently, actin aggregates form a 'corona' structure in the intercellular region between the egg and central cell where gametic fusion occurs. The corona may have a role in maintaining the close proximity of the egg and central cell and helping the two sperm cells move and bind to their target cells. The cytoskeleton may also be involved in causing the two nuclei of the egg and central cell to approach one another at the site of gametic fusion and transporting the two sperm nuclei into alignment with their respective female nucleus. The cytoskeleton is reorganised during early embryogenesis.

Publication Types:  PMID: 8081810 [PubMed - indexed for MEDLINE]

42. Effect of Plumbago zeylanica root powder induced preimplantationary loss and abortion on uterine luminal proteins in albino rats.

Devarshi P, Patil S, Kanase A.

Zoology Department, Shivaji University, Kolhapur, India.

P. zeylanica treatment during first 7 days of pregnancy abolished uterine proteins of 13,000, 19,000 and 26,000 and 75,000 Da molecular weights resulting in preimplantationary loss. Proteins having molecular weights 55,000 and 65,000 Da were absent in aborted rats, that were given P. zeylanica root powder since day 6 to day 17 of pregnancy. The results suggest that proteins having molecular weights 13,000, 19,000, 26,000 and 75,000 Da influence the implantation and proteins of 55,000 and 65,000 Da are required for the maintenance of the pregnancy.

PMID: 1889824 [PubMed - indexed for MEDLINE]

43. Hypolipidaemic and antiatherosclerotic effects of plumbagin in rabbits.

Sharma I, Gusain D, Dixit VP.

Department of Zoology, University of Rajasthan, Jaipur.

Plumbagin (2-methyl-5-hydroxy, 1:4 naphthoquinone) isolated from the roots of Plumbago zeylanica when administered to hyperlipidaemic rabbits, reduced serum cholesterol and LDL-Chol. by 53 to 86 percent and 61 to 91 percent respectively. It lowered cholesterol/phospholipid ratio by 45.8 percent and elevates the decreased HDL-Chol significantly. Further, Plumbagin treatment prevented the accumulation of cholesterol and triglycerides in liver and aorta and regressed atheromatous plaques of thoracic and abdominal aorta. Plumbagin treated hyperlipidaemic subjects excreted more fecal cholesterol and phospholipids. In conclusion-Plumbagin feeding brings about a definite regression of atheroma and prevents the accumulation of cholesterol and triglycerides in liver and aorta.

PMID: 1917004 [PubMed - indexed for MEDLINE]

44. Effects of plumbagin on antibiotic resistance in bacteria.

Durga R, Sridhar P, Polasa H.

Department of Microbiology, Osmania University, Hyderabad.

Plumbagin, a compound derived from the roots of Plumbago zeylanica (Chitramool) was studied for its effect on the development of antibiotic resistance using antibiotic sensitive strains of Escherichia coli and Staphylococcus aureus. A delayed growth was seen when these organisms were inoculated into the antibiotic (streptomycin/rifampicin) medium, due to development of resistance in some of the cells. However, the growth was completely prevented when the bacteria were grown in the medium containing antibiotic and plumbagin together, and this was attributed to prevention of development of antibiotic resistant cells.

Publication Types: PMID: 2188907 [PubMed - indexed for MEDLINE]

45. Isolation of Fixed and Viable Eggs, Central Cells, and Embryo Sacs from Ovules of Plumbago zeylanica.

Huang BQ, Russell SD.

Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma 73019.

Three alternative protocols for light microscopy, electron microscopy, and biochemical characterization of isolated megagametophytic tissues are described employing enzymic maceration and microdissection of living and fixed ovular tissue of Plumbago zeylanica. Morphologically well preserved megagametophytes are obtained using fixed ovules in two different regimes (nearly 40 and 60% yield, respectively). Fluorescein diacetate-positive megagametophytic cells are recovered in nearly 20% of unfixed ovules using the third regime.

PMID: 16666774 [PubMed - as supplied by publisher]

PMCID: PMC1061665

46. Two-Dimensional Electrophoretic Studies of the Proteins and Polypeptides in Mature Pollen Grains and the Male Germ Unit of Plumbago zeylanica.

Geltz NR, Russell SD.

Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma 73019.

Three fractions (male germ unit [MGU]-rich, cytoplasmic-particulate and water-soluble proteins) were isolated from pollen of Plumbago zeylanica L. Proteins were extracted using a phenol procedure and polypeptide patterns were compared on one- and two-dimensional polyacrylamide gels. The MGU-rich fraction contains the sperm and vegetative nucleus of the pollen grain and yielded 427 spots >33 kilodaltons. The cyto-plasmic-particulate fraction contained 515 spots >33 kilodaltons. The third fraction consisted of water-soluble proteins and polypeptides from the pollen cytoplasm, in which 285 spots (>33 kilodaltons) were identified. Of 133 polypeptide spots suitable for comparison, 18 were unique to the MGU-rich fraction, 3 to the cytoplasmic-particulate fraction, 14 to the water-soluble fraction, 65 were common to two different fractions (and absent in one), and 33 were common to all three of the fractions examined.

PMID: 16666380 [PubMed - as supplied by publisher]

47. Isolation of Sperm Cells from the Pollen of Plumbago zeylanica.

Russell SD.

Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma 73019.

Intact sperm cells of Plumbago zeylanica were released from mature pollen grains near anthesis using osmotic shock with 20% sucrose. Sperm cell yields of up to 75% can be attained by differential centrifugation using a clinical centrifuge with concentrations of up to 8.8 x 10(6) cells/milliliter. Such ;gametoplasts' remain intact for up to 24 hours according to Evan's blue exclusion and can be used for characterization or physiological manipulation.

PMID: 16664799 [PubMed - as supplied by publisher]

PMCID: PMC1075328

48. Preferential fertilization in Plumbago: Ultrastructural evidence for gamete-level recognition in an angiosperm.

Russell SD.

Department of Botany and Microbiology, University of Oklahoma, Norman, OK 73019.

Gametic fusion patterns in the angiosperm Plumbago zeylanica were determined by using cytoplasmically dimorphic sperm cells differing in mitochondrion and plastid content and then identifying paternal organelles through their ultrastructural characteristics within the maternal cytoplasm at the time of fertilization. The virtual absence of plastids within the sperm cell that is physically associated with the vegetative nucleus allows paternal plastids to be used to trace the fate of the two male gametes after fusion. Such paternal plastids were present in the egg in >94% of the observed cases, indicating the preferential fusion of the plastid-rich, mitochondrion-poor sperm cell with the egg. In only one instance did the opposite pattern occur. Since the possibility of this result occurring as the consequence of chance in random fusions is <1 in 7000, this represents strong evidence for the presence of a final putative recognition event occurring at the gametic level.

PMID: 16593605 [PubMed - as supplied by publisher]

 

49. Folklore information from Assam for family planning and birth control.

Tiwari KC, Majumder R, Bhattacharjee S.

PIP: The author collected folklore information on herbal treatments to control fertility from different parts of Assam, India. Temporary methods of birth control include Cissampelos pareira L. in combination with Piper nigrum L., root of Mimosa pudica L. and Hibiscus rosa-sinensis L. Plants used for permanent sterilization include Plumbago zeylanica L., Heliotropium indicum L., Salmalia malabrica, Hibiscus rosa-sinensis L., Plumeria rubra L., Bambusa rundinacea. Abortion is achieved through use of Osbeckia nepalensis or Carica papaya L. in combination with resin from Ferula narthex Boiss. It is concluded that there is tremendous scope for the collection of folklore about medicine, family planning agents, and other treatments from Assam and surrounding areas. Such a project requires proper understanding between the survey team and local people, tactful behavior, and a significant amount of time. Monetary rewards can also be helpful for obtaining information from potential respondents.

PMID: 12266264 [PubMed - indexed for MEDLINE]

50. Research on plants for fertility regulation in India.

Kamboj VP, Dhawan BN.

PIP: This present review of Indian plants investigated for fertility regulation includes published literature of the country and unpublished data of the Central Drug Research Institute (CDRI), located in Lucknow, India. Publications without supportive experimental data have not been included. It is evident from the data presented in the tables that most of the investigators have failed to include the valuable information on the time and place of collection and proper botanical authentication, if conducted, in their publications. The plants evaluated at the Institute do contain this information and their herbaria sheets are available at CDRI. The plants, with part used, type of extract, isolated compound/chromatographic fraction, dose, route and schedule of administration with animal used, and percentage activity are given in tables. The plants are classified according to their activity profile and presented accordingly. Plants for which the hormonal profile or toxicity data have been reported are dealt with under each type of activity. Most of the investigators did not develop the active plants, probably because of inconsistent results in repeat tests or lack of facilities. Major attention has been devoted to identifying plants with interceptive properties. The schedules used are more or less uniform and acceptable. On the basis of preliminary toxicity data, extracts/compounds from "Aristolochia indica," "Artemisia scoparia," "Hibiscus rosa sinensis," "Laccardia lacca," and "Plumbago zeylanica" exclude themselves from consideration for follow-up. Wherever done, the hormonal profiles revealed estrogenic activity in active extracts/fractions/compounds from "Artabotrys odoratissimus," "Datura quercifolia," "Daucus carota," "Embelia ribes," "Hibiscus rosa sinensis," "Pueraria tuberosa" and "Tabernaemontana heyneana." Thus they are not ideal for follow-up. Some more plants can be excluded initially because of low activity or equivocal reports on activity. The remaining plants, in order of priority, for follow-up should be "Ensete superbum," "Achyranthes aspera," "Lygodium flexosum," "Sapindus trifoliatus," "Polygonum hydropiper," and "Abrus precatorius." The next priority could be given to plants with weak estrogenicity. The CDRI has observed 100% anti-implantation activity by 4 plants in hamsters. These should be the potential plants for development since they appear to interfere with progesterone synthesis or utilization.

Publication Types: PMID: 6752588 [PubMed - indexed for MEDLINE]

51.Participation of Male Cytoplasm During Gamete Fusion in an Angiosperm, Plumbago zeylanica.

Russell SD.

The embryo sac of Plumbago, lacking synergids, reveals an alternative course for pollen tube growth in angiosperms and provides ultrastructural evidence for transmission of sperm cytoplasm into the zygote and endosperm. Study of such evolutionarily reduced female gametophytes may aid in elucidating the structural basis for genetic transmission of plastids and mitochondria in flowering plants.

PMID: 17741287 [PubMed - as supplied by publisher]

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