Neuropharmacological evaluation on four traditional herbs used as nervine tonic and commonly available as Shankhpushpi in India.

BACKGROUND: Shankhpushpi is an Ayurvedic drug, widely used for its actions on the central nervous system, especially to improve intellect and boost memory. Four botanicals viz. Canscora decussata Schult. (CD), Clitorea ternatea Linn. (CT), Convolvulus pluricaulis Choisy. (CP) and Evolvulus alsinoides Linn. (EA) are considered as sources of Shankhpushpi by Indian practitioners on the basis of their morphological descriptions given in ancient texts. OBJECTIVE: The present study was undertaken to evaluate the neuropharmacological effect of four herbs commonly identified as source of Shankhpushpi. MATERIALS AND METHODS: Methanol extracts of all four varieties were tested and evaluated in vitro and in vivo for their neuropharmacological effects. Experiments such as protection against ß-amyloid induced neurotoxicity on brain cell line (Neuro 2A), antioxidant potential, AchE (acetylcholinesterase enzyme) inhibition, and 5-LOX (lipoxygenase) enzyme inhibition were conducted for in vitro evaluation. For in vivo evaluation, scopolamine (0.3 mg/kg i.p.) induced memory retrieval using pole climbing apparatus and Morris water maze were performed in rat models. RESULTS: It was found that protective effects of EA and CD against ß-amyloid induced neurotoxicity in Neuro 2A cells were significantly higher than CT and CP. EA proved to be superior than other varieties on the basis of antioxidant activity, AchE inhibitory and LOX inhibitory activities. The preventive activity of EA on scopolamine induced memory retrieval in pole climbing and Morris water maze task in rats was found to be higher than that of CD, CT and CP. CONCLUSION: EA has remarkable neuropharmacological effect as compared to other three varieties of Shankhpushpi. This effect may be attributed due to the presence of steroids (stigmasterol and betulinic acid), coumarins (scopoletin) and flavonoids (ß-carotene and chlorogenic acid). Hence it can be used as a promising lead in development and management of neuronal disorders including Alzheimer's disease.

Antioxidant and hepatoprotective effects of mixed micellar lipid formulation of phyllanthin and piperine in carbon tetrachloride-induced liver injury in rodents.

Phyllanthin, a sparingly water-soluble hepatoprotective lignin obtained from Phyllanthus amarus Schum. et Thonn. (Euphorbiaceae) possesses low bioavailability. Phyllanthin along with piperine (a nutraceutical bioenhancer) was formulated as a mixed micellar lipid formulation (MMLF) in the present study and investigated to resolve the low bioavailability and enhance hepatoprotective effects on oral administration. Hepatoprotective, antioxidant and bioavailability studies of MMLF, a complex phosphatidylcholine formulation of phyllanthin (CP-PC), phyllanthin + piperine (CP-P-PC) and its corresponding non-formulated phyllanthin have been carried out. Phyllanthin (30 mg kg(-1) p.o.), CP-PC (30 mg kg(-1) p.o.), CP-P-PC (30 mg kg(-1) p.o.) and the reference drug silymarin (100 mg kg(-1), p.o.) were administered daily to rats for 10 days, followed by liver damage by administering a 1 : 1 (v/v) mixture of CCl4 and olive oil (1 ml kg(-1), i.p.) for 7 days from day 4 to day 10. The degree of protection was evaluated by determining the level of marker enzymes (SGOT and SGPT), bilirubin (TB) and total proteins (TP). Further, the effects of MMLF on lipid peroxidation (LPO), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GR) were estimated in liver homogenates to evaluate the antioxidant activity. Finally the concentration of phyllanthin was evaluated in plasma. EC50 values for the in vitro antioxidant assay with DPPH were found to be 19.99, 15.94 and 13.5 for phyllanthin, CP-PC and CP-P-PC, respectively. CP-P-PC (30 mg kg(-1) p.o.) showed significant (p < 0.05) hepatoprotective effect by reducing the levels of serum marker enzymes (SGOT, SGPT, and TB), whereas, elevated the levels of depleted total protein (TP), lipid peroxidation and antioxidant marker enzyme activities such as, GSH, SOD, CAT, GPX, and GR. The complex MMLF normalized adverse conditions of rat livers more efficiently than the non-formulated phyllanthin. The present findings indicate that the MMLF is helpful in solving the problem of low bioavailability of phyllanthin.

Protective effect of extracts of Baliospermum montanum (Willd.) Muell.-Arg. against paracetamol-induced hepatotoxicity-an in vivo and in vitro study.

AIM: Evaluation of hepatoprotective effect of ethyl methyl ketone and methanol sub-fractions obtained from methanol fraction of total methanol extract (ME) was carried out both in vivo and in vitro using paracetamol-induced toxicity. SETTINGS AND DESIGN: Hepatoprotective activity in vivo was assessed by determining the serum levels of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), alkaline phosphatase, total bilirubin, total cholesterol, total protein (TPTN), and albumin in serum. The studies were supported by histopathological examination of liver sections. In vitro activity was assessed by determining the change in hepatocyte viability and other parameters such as GOT, GPT and TPTN. MATERIALS AND METHODS: The ME of the roots of Baliospermum montanum was prepared and fractionated with chloroform and methanol to get activity guided fraction. The bio-active guided methanol fraction was further fractionated with ethyl methyl ketone and methanol. Both the fractions were evaluated for hepatoprotective activity against paracetamol-induced toxicity. STATISTICAL ANALYSIS: For determining the significant intergroup difference each parameter was analyzed separately, and one-way analysis of variance was carried out and the individual comparisons of the group mean values were done using Dunnet's test. RESULTS: Methanol sub-fraction prevented hepatic damage in vivo, induced by paracetamol, whereas both the sub-fractions showed hepatoprotective effect by restoring altered parameters in the selected in vitro model. The results were comparable with the standard hepatoprotective drug silymarin. CONCLUSIONS: This study underlines the therapeutic potential of B. montanum as per claims in Ayurveda in liver disorders.

Antidiabetic activity of alkaloids of Aerva lanata roots on streptozotocin-nicotinamide induced type-II diabetes in rats.

CONTEXT: The roots of Aerva lanata Linn. (Amaranthaceae) (AL) are employed traditionally as an antihyperglycaemic in the Ayurvedic system of medicine. OBJECTIVE: The present investigation is focus for identification and isolation of the bioactive compound from methanol roots extract of AL against streptozocin-nicotinamide induced elevated serum glucose level in rats. MATERIALS AND METHODS: The methanol extract of the roots was fractionated using different solvents. The partially purified alkaloid basified toluene fraction (PPABTF) showed the presence of alkaloids. The fraction (10 and 20 mg/kg) was tested for oral glucose tolerance test (OGTT) and in non-insulin-dependent diabetes mellitus (NIDDM)-induced elevated serum glucose level in rats. The fraction was also subjected to high performance thin layer chromatography (HPTLC) for the determination of content of individual alkaloids. RESULTS: Single oral administration of PPABTF (10 and 20 mg/kg) after 20 h caused a significant (p < 0.01) reduction in the serum glucose level (mg/dl). On other hand, PPABTF normalised plasma glucose levels after 2 weeks of repeated oral administration in diabetic rats (p < 0.01). HPTLC analysis on PPABTF showed the presence of three known alkaloids. The fraction was further subjected to column chromatography and the compounds identified by ultraviolet, infrared, mass spectroscopy and nuclear magnetic resonance, as canthin-6-one derivatives. CONCLUSION AND DISCUSSION: The PPABTF in the dose of 20 mg/kg showed significant effects on streptozotocin-nicotinamide induced type-II NIDDM in rats. The activity may be due to the presence of alkaloids like canthin-6-one derivatives.

Studies of acid resistance characteristics in multiple drug resistant Salmonella species isolated from tomatoes.

Salmonella species found to have a great potential of causing a variety of diseases ranging from gastroenteritis to enteric fever. Salmonella have been isolated from all food, animals and also found in the vegetables such as tomatoes, spinach etc. Several out breaks of Salmonellosis have been associated with the consumption of raw tomatoes. This is because of the fact that Salmonella attaches to the surface of tomatoes and also present in the interior part due to geotropic transmission via contaminated soil irrigated with contaminated water. .During the life cycle, Salmonella encounters the various environments such as acidic environment (low pH). To overcome such factors, Salmonella has certain adaptable mechanisms. In present 'study total 200 samples of tomatoes were analyzed out of which 10 samples were found to contain Salmonella. All the 10 isolates were then subjected to the antibiotic susceptibility testing and were found to be resistant against several antibiotics. These were subjected to acid resistant tolerance study.

Hepatoprotective potential of Tecomella undulata stem bark is partially due to the presence of betulinic acid.

ETHNOPHARMACOLOGICAL RELEVANCE: Tecomella undulata (TU;` Family Bignoniaceae) is used in Indian Ayurvedic system of medicine for treating various diseases including hepatic ailments. It is also incorporated in various marketed hepatoprotective polyherbal formulations. AIM: The present study was aimed at evaluating possible hepatoprotective role of isolated compounds from TU stem bark (TSB) using in vitro and in vivo experimental models. METHODS: In vitro cytotoxicity and hepatoprotective potential of various extract, fractions and isolated compounds from TU stem bark were evaluated using HepG2 cells. Rats were pre-treated with TU methanolic extract (TSB-7) or betulinic acid (MS-2) or silymarin for 7 days followed by a single dose of CCl(4) (0.5 ml/kg, i.p.). Plasma markers of hepatic damage, hepatic antioxidants and indices of lipid peroxidation along with microscopic evaluation of liver were assessed in control and treatment groups. RESULTS: TSB-2 and MS-1 accounted for significant cell death whereas; TSB-1, TBS-7, TSB-9, TSB-10 and, MS-2 did not register significant cytotoxicity. Further, non-cytotoxic components exhibited ascending grade of hepatoprotection in vitro (TSB-10

Amelioration of carbon tetrachloride induced hepatotoxicity in rats by standardized Feronia limonia. Linn leaf extracts.

The hepatoprotective potential of standardized Feronia limonia (Family, Rutaceae) methanolic extract (FL-7) and chloroform soluble fraction (FL-9) were assessed against carbon tetrachloride (CCl4) induced oxidative stress and hepatotoxicity in rats. Rats treated with CCl4 recorded significant elevation in plasma markers of hepatic injury, alteration in hepatic antioxidant status and histopathological damages. However, rats pretreated with FL-7 (200 or 400 mg/kg, p.o.) and FL-9 (100 or 200 mg/kg, p.o.) for 7 days and later administered CCl4 (0.5 ml/kg, i.p.) recorded lowered indices of the above mentioned parameters and minimal histological damage in a dose dependent manner. These results were comparable to that of CCl4+silymarin treated rats. The results obtained with FL-7 and FL-9 are attributable to their free radical scavenging potential due to high contents of polyphenols and flavonols recorded herein. Overall, this study establishes the efficacy of FL-7 and FL-9 as hepatoprotective agents against CCl4 induced hepatotoxicity in rats.

Hypoglycemic activity of C-glycosyl flavonoid from Enicostemma hyssopifolium.

CONTEXT: Enicostemma hyssopifolium Verdoon (Gentianaceae) has been documented for various therapeutic effects in traditional systems of medicine; the hypoglycemic and hypolipidemic activities are also well reported. OBJECTIVE: Bioactivity guided fractionation of methanol extract of E. hyssopifolium to test the hypothesis that E. hyssopifolium and its constituents influence cells and systemic glucose homeostasis. MATERIALS AND METHODS: Derived fraction and isolated compounds were studied for (1) aldose reductase (AR) inhibition, (2) α-glucosidase inhibition, (3) effect on gluconeogenesis in rat hepatoma, (4) cytoprotection against streptozotocin (STZ)-induced toxicity on RINm5F cells, (5) normalization of glycemic control in acute hyperglycemic rat model, and (6) insulin-releasing effect both in vitro and in vivo. RESULTS: The results indicated that E. hyssopifolium can modify the glucose homeostasis at the cellular level. Two bioactive constituents were identified. Swertisin was found to inhibit AR (IC(50) 1.23 µg/mL) and α-glucosidase (IC(50) 1.89 µg/mL). It also possessed a significant cytoprotective action of RINm5F cell line against toxicant STZ. Swertiamarin was found to have hepatic gluconeogenesis inhibiting and insulin-releasing effect on rat hepatoma and RINm5F cells, respectively. The results of the in vivo study showed that swertiamarin, unlike the in vitro effect, produced no significant raise of insulin secretion. Swertisin normalized the serum glucose 60 min after high dose of glucose (2 g/kg, i.p.) in rats. DISCUSSION AND CONCLUSION: These findings demonstrate that the fraction derived from the aerial part of E. hyssopifolium achieve normoglycemic status in hyperglycemic conditions via various mechanisms. The constituents swertiamarin and swertisin are responsible for bioactivity.

Cytotoxicity evaluation and hepatoprotective potential of bioassay guided fractions from Feronia Limmonia Linn leaf.

OBJECTIVE: To evaluate the cytotoxicity and hepatoprotective potentials of extracts, fractions or isolated compound from the leaves of Feronia limonia (F. limonia). METHODS: Qualitative phytochemical analysis of extracts, fractions or compound was performed by means of thin layer chromatography and spectroscopic assays. The % purity of compound was measured by analytical HPLC. Extracts, fractions or compound have been individually evaluated for their cytotoxicity effects (10, 20, 100, 250, 500, 750 and 1 000 µg/mL). Based on the inhibitory concentration (IC50) obtained from the cell viability assay, graded concentrations of extracts, fractions or isolated compound were assessed (10, 20, 50, 100, 200 µg/mL) for its hepatoprotective potential against CCl4-induced hepatotoxicity by monitoring activity levels of serum glutamatic pyruvatic transaminase (SGPT) and serum glutamic oxaloacetic transaminase (SGOT). RESULTS: Results indicated that the methanol extract of F. limonia was non-toxic and hepatoprotective in nature as compared with the petroleum ether extract. The acetone fraction of methanolic extract also showed similar properties but the subsequent two fractions were cytotoxic. However, the pure compound isolated from the penultimate fraction of methanolic extract was non-toxic and hepatoprotective in nature. Biochemical investigations (SGOT, SGPT) further corroborated these cytological observations. CONCLUSIONS: It can be concluded from this study that F. limonia methanol extract, some fractions and pure isolated compound herein exhibit hepatoprotective activity. However, cytotoxicity recorded in the penultimate fraction and investigation of structural details of pure compound warrants further study.

Phytopharmacologic aspects of Canscora decussata Roem and Schult.

Nature is an inexhaustible source of secondary metabolites-different types of alkaloids, terpenoids, phenolics, and other classes of organic compounds. In the process of isolation, purification and determination of the structures of lead, with their biological effectiveness, every type of experimental tool and strategy, known too and developed over the years by various practitioners. The present review is an attempt to compile information on various aspects of Canscora decussata, "Shankhpushpi" of Indian traditional system of medicine. The phytoconstituents, such as phenolic compounds, xanthones, and triterpenoids were isolated from different parts of the plant. The plant possesses immunomodulatory, analgesic, anticonvulsant, antitubercular, antiinflammatory, spermicidal, central nervous system-depressive, and cardiostimulant properties. Clinical trials of marketed formulation showed very encouraging results.

A simple and rapid method for simultaneous estimation of glycyrrhetinic Acid and piperine by hptlc in a herbomineral formulation.

Many of the traditional herbal formulations contain extracts of Piper longum and Glycyrrhiza glabra, piperine and glycyrrhetinic acid respectively, being active constituents of these two herbs. An attempt has been made to develop a simple, precise, rapid, and cost-effective high-performance thin-layer chromatographic (HPTLC) method for simultaneous estimation of these in a herbomineral formulation (Efiplus(®) Capsules). Precoated silica gel 60 F(254) plates with toluene-ethyl acetate-glacial acetic acid 12.5:7.5:0.5, as mobile phase were used in chromatographic determinations. The plates were scanned and the compounds were quantified at their wavelengths of maximum absorption of 260 and 331 nm for glycyrrhetinic acid and piperine respectively. The respective R(F), values of glycyrrhetinic acid and piperine were 0.51 and 0.55. Under these experimental conditions linearity was observed between 0.8-2.6 µg/ spot for glycyrrhetinic acid and between 10-50 ng/ spot for piperine and average recovery was 96.25% for glycyrrhetinic acid and 98.55% for piperine.

A comprehensive review on Nymphaea stellata: A traditionally used bitter.

Nymphaea stellata Willd. (Syn. Nymphaea nouchali Burman f.) (Nymphaeaceae) is an important and well-known medicinal plant, widely used in the Ayurveda and Siddha systems of medicines for the treatment of diabetes, inflammation, liver disorders, urinary disorders, menorrhagia, blenorrhagia, menstruation problem, as an aphrodisiac, and as a bitter tonic. There seems to be an agreement between the traditional use and experimental observations, such as, hepatoprotective, anti-inflammatory, and particularly antidiabetic activity. Nymphayol, a steroid isolated from the flowers has been scientifically proved to be responsible for the traditionally claimed antidiabetic activity; it reverses the damaged endocrine tissue and stimulates secretion of insulin in the ß-cells. However, taking into account the magnitude of its traditional uses, the studies conducted are still negligible. This review is an attempt to provide the pharmaceutical prospective of Nymphaea stellata.

Comparative pharmacognostical investigation on four ethanobotanicals traditionally used as Shankhpushpi in India.

People in Indian region often apply Shankhpushpi and other Sanskrit-based common name to Evolvulus alsinoides, Convolvulus pluricaulis, Canscora decussata, and Clitorea ternatea. These are pre-European names that are applied to a medicinal plant. Before the establishment of British rule, like the other books, ayurvedic treatises were also hand written. This might be one of the reasons due to which ayurveda could not stand parallel to the western medicine and an ambiguity is reflected in the interpretation of names and description of drugs found in the books like Charaka Samhita and Sushruta Samhita. The most widespread application of Shankhpushpi is for mental problems, but they have been considered for an array of other human maladies. The present investigation deals with the comparative pharmacognostical evaluation of four ethanobotanicals of Shankhpushpi. A comparative morphoanatomy of the root, stem, and leaves has been studied with the aim to aid pharmacognostic and taxonomic species identification. Various physicochemical, morphological, histological parameters, comparative high-performance thin-layer chromatography (HPTLC), and comparative high-performance liquid chromatography (HPLC), chromatogram of methanolic extract presented in this communication may serve the purpose of standard parameters to establish the authenticity of commercialized varieties and can possibly help to differentiate the drug from the other species. All the parameters were studied according to the WHO and pharmacopoeial guidelines.

Protective effect of stem bark of Ceiba pentandra linn. against paracetamol-induced hepatotoxicity in rats.

The present study reports protective activity of ethyl acetate fraction of methanol extract of stem bark of Ceiba pentandra against paracetamol-induced liver damage in rats. The ethyl acetate fraction (400 mg/kg) was administered orally to the rats with hepatotoxicity induced by paracetamol (3 gm/kg). Silymarin (100 mg/kg) was used as positive control. High performance thin layer chromatography (HPTLC) fingerprinting of ethyl acetate fraction revealed presence of its major chemical constituents. A significant (P < 0.05) reduction in serum enzymes GOT (ALT), aspartate aminotransferase (AST), GPT alkaline phosphatase (ALP), total bilirubin content and histopathological screening in the rats treated gave indication that ethyl acetate fraction of methanolic extract of Ceiba pentandra possesses hepatoprotective potential against paracetamol-induced hepatotoxicity in rats.

Hepatoprotective effect of Pergularia daemia (Forsk.) ethanol extract and its fraction.

Ethanol extract and its ethanol fraction from aerial parts of P. daemia exhibited significant hepatoprotective effect against CCl4 induced hepatotoxicity in rats. Glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, alkaline phosphatase, total bilirubin, total cholesterol, total protein and albumin in serum indicated hepatoprotective effect of the ethanol extract and its ethanol fraction. Histopathological examination of liver sections confirmed that, pre-treatment with ethanol extract and its ethanol fraction prevented hepatic damage induced by CCl4. The results were comparable with the standard hepatoprotective drug silymarin. The extract and its fraction showed no signs of toxicity up to a dose level of 2000 mg/kg. It is suggested that, the presence of flavonoids in ethanol extract and its ethanol fraction may be responsible for hepatoprotective properties. High Performance Thin Layer Chromatography profile of flavonoids of bio-active extracts was developed using quercetin-3-glucoside as a marker. Results indicate hepatoprotective properties of ethanol extract of P. daemia.

Simultaneous estimation of andrographolide and wedelolactone in herbal formulations.

Andrographolide and wedelolactone are active components of Andrographis paniculata and Eclipta alba, respectively. The extracts of these plants are used in many traditional hepatoprotective formulations. An attempt has been made to develop an accurate, precise and specific HPTLC method to quantify simultaneously both these chemical markers of diversified chemical structures in different dosage forms like tablet and syrup. Precoated silica 60F(254) plates with toluene:acetone:formic acid (9:6:1) as mobile phase and detection wavelength of 254 nm were used. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 200 to 400 ng/spot for andrographolide and 100 to 200 ng/spot for wedelolactone. The limit of detection and the limit of quantification for andrographolide were 26.16 and 79.28 ng/spot, respectively and for wedelolactone 5.06 and 15.32 ng/spot, respectively.

Immunomodulatory activity of petroleum ether extract of flower heads of Sphaeranthus indicus Linn.

The petroleum ether extract from the flower heads of Sphaeranthus indicus Linn. was found to be effective in increasing phagocytic activity, hemagglutination antibody titer and delayed type hypersensitivity when tested in mice. Activity of the petroleum extract was tested at five different dosing levels to establish a dose-response relationship. It was found that 200 mg/kg dose was the optimum dose, and at higher doses the activity was either reduced or showed no further increase. The present study, therefore, reveals that the drug shows good promise as an immunomodulatory agent, which acts by stimulating both humoral and cellular immunity as well as phagocytic function.

Evaluation of free radical scavenging activity of Butea monosperma Lam.

In the present study, ethyl acetate, butanol and aqueous fractions derived from total methanol extract of Butea monosperma flowers were evaluated for radical scavenging activities using different in vitro models like reducing power assay, scavenging of 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, hydroxyl radical and inhibition of erythrocyte hemolysis using 2, 2' azo-bis (amidinopropane) dihydrochloride (AAPH). Methanol extract along with its ethyl acetate and butanol fractions showed potent free radical scavenging activity, whereas aqueous fraction was found to be devoid of any radical scavenging properties. The observed activity could be due to the higher phenolic content in the extracts (16.1, 25.29, and 17.74% w/w in methanol extract, ethyl acetate and butanol fractions respectively). HPTLC fingerprint profile of the ethyl acetate and butanol fractions were developed which would serve as reference standard for quality control of the extracts.

Hepatoprotective effect of extracts from Pergularia daemia Forsk.

Pergularia daemia (Asclepiadaceae) is a perennial herb growing widely along the road sides of India. It has been used in folk medicine for the treatment of liver disorders. The aim of this work is to study the hepatoprotective effect of crude ethanolic and aqueous extracts from the aerial parts of Pergularia daemia. The aqueous and ethanolic extracts obtained from aerial parts of Pergularia daemia were evaluated for hepatoprotective activity in rats by inducing liver damage by carbon tetrachloride. The ethanolic extract at an oral dose of 200 mg/kg exhibited a significant (P<0.05) protective effect by lowering serum levels of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, alkaline phosphatase, total bilirubin and total cholesterol and increasing the levels of total protein and albumin levels as compared to silymarin used as a positive control. These biochemical observations were supplemented by histopathological examination of liver sections. The activity may be a result of the presence of flavonoid compounds. Furthermore, the acute toxicity of the extracts showed no signs of toxicity up to a dose level of 2000 mg/kg. Thus it could be concluded that ethanolic extract of Pergularia daemia possesses significant hepatoprotective properties.

Immunostimulatory effect of methanol extract of Curculigo orchioides on immunosuppressed mice.

The methanol extract of rhizomes of Curculigo orchioides Gaertn. (Amaryllidaceae) holds potential as a protective agent against cytotoxic drugs. The extract when studied on humoral and cell mediated immunity in normal, as well as cyclophosphamide-induced immunosuppressed mice produced an increase in humoral antibody (HA) titre, delayed type hypersensitivity (DTH) and levels of WBC in a dose dependent manner.