The constituents in the roots of Plumbago auriculata Lam. and Plumbago zeylanica L. responsible for antibacterial activity.
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Africa, Southern↗
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Protective effect of Plumbago zeylanica against cyclophosphamide-induced genotoxicity and oxidative stress in Swiss albino mice.
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.
Animals↗
An investigation into the biochemical basis of the observed hyperglycaemia in rats treated with ethanol root extract of plumbago zeylanica.
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.
Animals↗
Isolation of secondary metabolites from roots of Plumbago auriculata Lam. by countercurrent chromatography.
Centrifugal liquid-liquid partition chromatography presents significant advantages for the separation and purification of plant metabolites owing to the short operational time of the process and the elimination of possible irreversible adsorption of compounds. The crude chloroform extract from roots of Plumbago auriculata was analysed by countercurrent chromatography using hexane:ethyl acetate:methanol:water (40:10:10:2, v/v) as solvent system. The isolation of the naphthoquinones plumbagin and epi-isoshinanolone, the steroids sitosterol and 3-O-glucosylsitosterol, plumbagic and palmitic acids was easily achieved. Naphthoquinones are typical components of Plumbago species and they show interesting biological activities.
Brazil↗
Behavior of plastid nucleoids during male gametogenesis in Plumbago auriculata.
We characterized the behavior of plastid (pt) and mitochondrial (mt) nucleoids during male gametogenesis in Plumbago auriculata in three dimensions. The behavior of pt-nucleoids and mt-nucleoids differed throughout male gametogenesis. Pt-nucleoids were distributed in a characteristic manner in three stages: in the early microspore, pt-nucleoids assemble around cell nucleus; in the mid-generative cell, pt-nucleoids gather at the internal side of the pollen; in the late-generative cell, pt-nucleoids aggregation turns its pole to the external side of the pollen. We also studied organelle nucleoids in the egg and the central cell by a method in which semi-thick sections of resin-embedded anthers and ovaries were observed by confocal laser scanning microscopy. The number of pt-nucleoids in the sperm cell did not differ significantly from that in the egg. These results suggest that the behavior of DNA-containing organelles is regulated strictly during male gametogenesis in P. auriculata, and that a biparental inheritance of plastids in the Plumbago embryo is more favored than was previously thought.
Cell Nucleus↗
Plumbagin quantification in roots of Plumbago scandens L. obtained by different extraction techniques.
The Plumbago genus belongs to the Plumbaginaceae family and it is known due to its variety of biological uses, most of them attributed to the presence of naphthoquinones. Plumbagin is a naturally occurring naphthoquinone that can be obtained from roots of Plumbago scandens L. In order to find out the better technique for plumbagin extraction, were applied: static maceration, dynamic maceration, with assistance of ultrasonic waves and in Soxhlet apparatus. Four compounds were qualitatively detected in all extracts: the naphthoquinones plumbagin and epi-isoshinanolone, palmitic acid and sitosterol. Plumbagin was always the major component in all analyzed extracts and it was quantitatively determined by gas chromatograph coupled with mass spectrometer. Soxhlet was the most efficient extraction technique however, prolonged heating time promoted plumbagin degradation.
Chromatography, Gas↗
Intoxication of goats by Plumbago scandens in Northeastern Brazil.
In the State of Paraiba, Northeastern Brazil, goat mortality occurred with the ingestion of Plumbago scandens. The fresh plant was then given experimentally to 4 goats at 5, 10, 17.5 or 25 g/kg bw. Depression, anorexia, salivation with foamy saliva, bellowing, bruxism, humpbacked posture, bloat, ruminal atony, continuous lateral head movements, tachycardia, dyspnea and dark brown to black urine were observed in the goats given 17.5 or 25 g/kg bw. The goats receiving 5 or 10 g/kg bw had less severe signs. The goat dosed with 25 g/kg bw died after 18-20 h. All others recovered in 3-9 d. At necropsy of the high dose goat, the main lesions were dark violet to black discoloration of the mucosa of the tongue, esophagus, reticulum and ventral sac of the rumen, and gelatinous edema in the visceral ruminal peritoneum. Histologically the reticulum and ventral rumen sac had diffuse epithelial necrosis and severe edema and neutrophilic infiltration of the submucosa. Separation of the ruminal epithelium from the submucosa was observed. Epithelial degeneration and necrosis was also seen in the omasum, esophagus and tongue. Reproduction of the disease with clinical signs similar to those observed by the farmer in the spontaneously affected goats suggests that the clinical mortality was caused by ingestion of Plumbago scandens.
Animals↗
Antimycobacterial constituents from Juniperus procera, Ferula communis and Plumbago zeylanica and their in vitro synergistic activity with isonicotinic acid hydrazide.
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.
Antitubercular Agents↗
Central nervous system stimulatory action from the root extract of Plumbago zeylanica in rats.
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.
Animals↗
UV/Vis spectra and solubility of some naphthoquinones, and the extraction behavior of plumbagin from Plumbago scandens roots in supercritical CO2.
The solubility of 1,4-naphthoquinone, plumbagin, lawsone, and juglone in supercritical carbon dioxide was determined spectroscopically at 40 degrees C, and in the pressure range 8-18 MPa. Their solubilities at 12 MPa were between 0.3 and 10 g L(-1). Plumbagin from Plumbago scandens L. roots was extracted at 40 degrees C and 20 MPa. The extracted plumbagin mass fraction was up to 0.2% in fresh roots but down to about 0.006% in aged roots. n-Hexane and chloroform extraction of such aged roots indicates that the older and dryer the roots are, the stronger they bind plumbagin. Reversed-phase HPLC indicated a relatively pure plumbagin extract with supercritical carbon dioxide.
Carbon Dioxide↗
A highly efficient in vitro plant regeneration system and Agrobacterium-mediated transformation in Plumbago zeylanica.
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.
Cotyledon↗
Toxic effects of crude root extract of Plumbago rosea (Rakta chitraka) on mice and rats.
The ethanolic root extract of Plumbago rosea (Plumbaginaceae) was studied for acute toxicity in mice and subacute toxicity in rats. The 24 h LD50 values of the extract in mice were 239.88 mg and 1148.15 mg/kg b.wt. for intraperitoneal and oral routes, respectively. Oral administration of doses above 1250 mg/kg produced severe diarrhea. In subacute toxicity studies no mortality was observed when 50 mg/kg of the extract was injected i.p. daily for 30 days; however, there was no weight gain in the treated rats. Significant reduction in the weights of liver, kidney, thymus and testes was observed in the male rats, while the spleen weight showed a significant increase from control. The females showed a significant loss in thymus weight and a gain in the weight of the uterus, but the liver and spleen did not show any weight change from the control. There was a significant increase in total WBC and neutrophil counts as well as in the levels of serum alkaline phosphatase and alanine transaminase (ALT) in both sexes. Similarly, the liver alkaline phosphatase level was significantly higher than control, but a significant reduction was observed in the DNA, RNA and total proteins. Thus, a higher drug dose (total dose of 15 g/kg b.wt.) was tolerated in fractionated administration, but it had a growth inhibitory effect in both sexes. The males appear to be more susceptible than females when individual organs are considered.
Alanine Transaminase↗
Determination and identification of plumbagin from the roots of Plumbago zeylanica L. by liquid chromatography with tandem mass spectrometry.
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.
Chromatography, Liquid↗
High-performance liquid chromatography for quantification of plumbagin, an anti-Helicobacter pylori compound of Plumbago zeylanica L.
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.
Chromatography, High Pressure Liquid↗
Anti-Helicobacter pylori activity of Plumbago zeylanica L.
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.
Anti-Bacterial Agents↗
Cytotoxic constituents from Plumbago zeylanica.
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).
Antineoplastic Agents, Phytogenic↗
Antiparkinsonian-like effects of Plumbago scandens on tremorine-induced tremors methodology.
Tremorine-induced tremors model is used to evaluate antiparkinsonian drugs because rest tremor is a sign that distinguishes Parkinson's disease (PD) from other diseases. The effects of crude ethanolic extract (CEE) and total acetate fraction (TAF) of Plumbago scandens were investigated at several doses. These extracts at doses of 125 and 250 mg/kg i.p. failed to reduce tremors in tremorine-treated mice. TAF showed significant effects only at a dose of 500 mg/kg. Both CEE and TAF at doses of 1000 and 2000 mg/kg i.p. suppressed the tremors in a dose-dependent fashion for 60 min. Biperiden, an anticholinergic drug, was used as standard at a dose of 3 mg/kg i.p. This study suggests that P. scandens is a plant with possible therapeutic value for PD.
Animals↗
Plumbago zeylanica action on blood coagulation profile with and without blood volume reduction.
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.
Animals↗