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.

Sphaeranthus indicus induces apoptosis through mitochondrial-dependent pathway in HL-60 cells and exerts cytotoxic potential on several human cancer cell lines.

PURPOSE: The study was designed to screen Sphaeranthus indicus, Ganoderma lucidum, and Urtica dioica for their anticancer activity against human cancer cell lines. Phytochemical screening of active extracts was also planned. METHODS: Petroleum ether, ethanolic, and aqueous extracts of S indicus Linn, G lucidum P Karst, and U dioica Linn were subjected to cytotoxicity studies using 7 different cancer cell lines. Potent cytotoxicity was noted in petroleum ether extract of S indicus (SIP), which inhibited proliferation of various cancer cell lines. Growth inhibition was determined by sulforhodamine B assay. Two biochemical markers, namely ß-sitosterol and 7-hydroxyfrullanolide were isolated and characterized using high-performance thin layer chromatography, melting point, Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, and mass analysis. Cytotoxicity of isolated ß-sitosterol and 7-hydroxyfrullanolide were also determined. The IC(50) of SIP was calculated in the HL-60 cells and was found to be 53 µg/mL. Furthermore, SIP induced apoptosis in human leukemia HL-60 cells as measured by several biological end points. Cell cycle analysis and change in mitochondrial membrane potential was quantified by flow cytometry. Subsequently, using annexin V/PI assay, proportion of cells actively undergoing apoptosis was determined. Changes in DNA were observed by DNA ladder assay. RESULTS: SIP induced apoptotic bodies formation, induced DNA laddering, enhanced annexin-V-FITC binding of the cells, increased sub-G(0) DNA fraction, and induced loss of mitochondrial membrane potential (ΔΨm) in HL-60 cells. SIP also elevated the caspase 3 and caspase 9 levels in the HL-60 cells, which clearly indicates the involvement of the intrinsic proteins in inducing apoptosis. DISCUSSION: All the above parameters revealed that SIP induced apoptosis through the mitochondrial-dependent pathway in HL-60 cells. The criterion for anticancer activity in cytotoxicity assay was ≥70% growth inhibition at 100 µg/mL against at least 4 cell lines. As G lucidum and U dioica did not exhibit appreciable inhibitory activity against human cancer cell lines (less than 50%), they were not included in the study thereafter. The results established that SIP has apoptosis-inducing effect against HL-60 cells in vitro and is a promising candidate for further anticancer study. ß-Sitosterol and 7-hydroxyfrullanolide can be considered to be potent anticancer compounds isolated from SIP on the basis of present studies.

Effect of Tephrosia purpurea (L.) Pers. Leaves on Gentamicin-Induced Nephrotoxicity in Rats.

The aim of the study was to evaluate the nephroprotective and nephrocurative effects of Tephrosia purpurea (L.) Pers. leaves against gentamicin-induced acute renal injury in albino rats. The maximum free radical scavenging activity of the ethanolic extract was the basis for the selection of this extract for the in vivo study. Gentamicin (40 mg/kg, s.c.) was administered to induce toxicity in the toxic group and the ethanolic extract (200 mg/kg p.o.) was administered in all treated groups. Blood urea and serum creatinine levels were monitored to assess the effects. The antioxidant potential was also evaluated by the estimation of reduced glutathione (GSH) and malondialdehyde (MDA). Gentamicin intoxication caused significant increases in blood urea and serum creatinine levels as compared to the normal control. In the preventive regimen, the extract (200 mg/kg, p.o.) showed significant reductions in the elevated blood urea and serum creatinine. Histopathological changes were in accordance with the biochemical findings. Also in the curative regimen, the blood urea and serum creatinine levels revealed significant curative effects. In our in vivo antioxidant activity, the GSH level was significantly (P< 0.05) increased in the extract-treated groups, whereas MDA was reduced significantly (P< 0.05). Further thin layer chromatography (TLC) and high-performance thin layer chromatography (HPTLC) led us to ascertain the presence of rutin and quercetin in the extract. We were able to isolate and characterize an isolate from the ethanolic extract and characterize it on the basis of chromatographic, melting point, FTIR, NMR, and mass spectroscopic studies. The findings suggest that the ethanol extract of Tephrosia purpurea leaves possesses marked nephroprotective and curative activities without any toxicity. The proposed mechanisms for the claimed activity are antioxidant activity and the inhibition of an overproduction of NO and Cox-2 expression. These activities may be attributed to the presence of phenolics and flavonoidal compounds like rutin and quercetin. Thus, it can be said that Tephrosia purpurea could offer a promising role in the treatment of acute renal injury caused by nephrotoxins like gentamicin.

An update on Murraya koenigii spreng: a multifunctional Ayurvedic herb.

Murraya koenigii Spreng (Rutaceae), a medicinally important herb of Indian origin, has been used for centuries in the Ayurvedic system of medicine. Leaves, fruits, roots and bark of this plant are a rich source of carbazole alkaloids. These alkaloids have been reported for their various pharmacological activities such as antitumor, antiviral, anti-inflammatory, antidiarrhoeal, diuretic and antioxidant activities. Apart from these activities, the plant is reported to possess a wide spectrum of biological activities. Phytochemistry and pharmacology of this plant make a demand of an exhaustive review of its potential as a valuable therapeutic agent for the treatment and management of various ailments frequently affecting humans. The present review gives a detailed description of the phytochemical, pharmacological, clinical and pre-clinical works carried out on this medicinal herb and also throws light on its therapeutic potential.

Sphaeranthus indicus attenuates testosterone induced prostatic hypertrophy in albino rats.

The present study reports the attenuating effect of Sphaeranthus indicus extracts (SI) on prostatic hyperplasia induced by testosterone in albino rats. In vitro studies were conducted to assess the 5α-reductase inhibitory potential of the petroleum ether, ethanolic and aqueous extracts of SI. A biochemical marker, ß-sitosterol, was isolated and extracts were characterized utilizing HPTLC. Testosterone (3 mg/kg s.c.) was administered to the rats along with the test extracts and isolated ß-sitosterol for a period of 28 days. The weight of the rats, the urine output, serum testosterone concentrations and prostate-specific antigen (PSA) levels were recorded. The prostate/body weight ratio (P/BW) was calculated and histological studies were performed to observe the changes in the histoarchitecture of the prostate. Finasteride was used as a positive control (1 mg/kg p.o.). Sphaeranthus indicus extracts attenuated the increase in the P/BW ratio induced by testosterone in the treated groups. The petroleum ether extract exhibited the best activity, although the ethanol and aqueous extracts also exhibited significant activity. Urine output was also improved significantly, demonstrating the clinical implications of the study. Histological studies, testosterone levels which were measured weekly and PSA levels measured at the end of the study also support claims for the potential use of Sphaeranthus indicus in the treatment of prostatic hyperplasia.

Effects of Phyllanthus acidus (L.) Skeels fruit on carbon tetrachloride-induced acute oxidative damage in livers of rats and mice.

OBJECTIVE: The present study was undertaken with a view to validate the traditional use of Phyllanthus acidus (L.) Skeels fruit as a hepatoprotective agent. METHODS: The 70% ethanolic extract of P. acidus fruit (100, 200 and 400 mg/kg, p.o.), and reference drug silymarin (100 mg/kg, p.o.) were given to rats of different groups respectively once a day for 5 d and the carbon tetrachloride (CCl4) (2 mL/kg, subcutaneously) was given on days 2 and 3. Serum levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin (TB) and total protein (TP) were assessed along with liver histopathological examination. The effects on oxidative stress markers such as lipid peroxidation (LPO), reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were also assessed in liver tissue homogenate to evaluate in vivo antioxidant activity. In addition, the effects on hexobarbitone-induced sleeping time were observed and the free radical-scavenging potential was determined by using 2, 2-diphenyl-1-picrylhydrazil (DPPH) in mice. RESULTS: P. acidus extracts and silymarin exhibited a significant hepatoprotective effect as evident from the decreases of serum AST, ALT and ALP levels and LPO and increases in the levels of TP, GSH, SOD, CAT, and GPx compared with control group (P<0.01 or P<0.05). The biochemical results were supplemented with results of histopathological sections of the liver tissues. P. acidus extracts considerably shortened the duration of hexobarbitone-induced sleeping time in mice compared with control group (P<0.01) and showed remarkable DPPH-scavenging activity. CONCLUSION: The present findings suggest that the hepatoprotective effect of P. acidus against CCl4-induced oxidative damage may be related to its antioxidant and free radical-scavenging potentials.

Effect of Evolvulus alsinoides Linn. on learning behavior and memory enhancement activity in rodents.

In the Ayurvedic system of medicine, the whole herb of 'Shankhpushpi' has been employed clinically for centuries for its memory potentiating, anxiolytic and tranquilizing properties. The present study was undertaken to investigate the effects of Evolvulus alsinoides (EA), considered as Shankhpushpi on learning and memory in rodents. Nootropic activity using Cook and Weidley's pole climbing apparatus, passive avoidance paradigms and active avoidance tests were used to test learning and memory. The ethanol extract of EA and its ethyl acetate and aqueous fractions were evaluated for their memory enhancing properties. Two doses (100 and 200 mg/kg p.o.) of the ethanol extract and ethyl acetate and aqueous fractions were administered in separate groups of animals. Both doses of all the extracts of EA significantly improved learning and memory in rats. Furthermore, these doses significantly reversed the amnesia induced by scopolamine (0.3 mg/kg i.p.). Nootropic activity was compared using piracetam as the standard. EA also exhibited potent memory enhancing effects in the step-down and shuttle-box avoidance paradigms.

Effect of Benincasa hispida fruits on testosterone-induced prostatic hypertrophy in albino rats.

BACKGROUND: Benincasa hispida Cogn. has been used traditionally in India for the management of urinary disorders. The fruit of B hispida is used as a diuretic and the seeds have been reported to possess antiangiogenic effects in prostate cells. OBJECTIVE: The aim of the present study was to examine the effect of petroleum ether extract, ethanolic extract, and B hispida seed oil on hyperplasia of the prostate induced by the subcutaneous administration of testosterone in rats. METHODS: In vitro studies were performed to determine the 5α-reductase inhibitory potential of the extracts. The results of those studies paved the way for the pharmacologic screening of the extracts to assess their potential against testosterone-induced hyperplasia in rats. Nine groups containing 10 rats per group were created for this study. Hyperplasia was induced by administration of testosterone (3 mg/kg SC) for 14 days in all the groups except the vehicle-treated group. Simultaneous administration of petroleum ether extract (100 or 200 mg/kg PO), ethanolic extract (100 or 200 mg/kg PO), and B hispida seed oil (20 or 40 mg/kg PO) was conducted. A standard 5α-reductase inhibitor (ie, finasteride) was used as a positive control. The weight of the rats was recorded on day 0 (ie, day 1 of the study) and on day 14, and the influence of testosterone and test extracts on the weight of the rats was determined. On day 14, rats were euthanized; prostates were dissected out, and weighed. The rats' prostate/body weight (P/BW) ratio was then determined. Histologic examinations were performed on prostates from each group. RESULTS: The petroleum ether extract as well as B hispida seed oil exhibited inhibition of 5α-reductase activity in in vitro studies. Ethanolic extract did not exhibit significant inhibitory potential in vitro. Further in vivo study found that testosterone treatment significantly increased the rats' P/BW ratio in all the groups except the vehicle-treated rats, and this increase in weight was significantly inhibited in rats administered petroleum ether extract (100 and 200 mg/kg PO) and B hispida seed oil (20 and 40 mg/kg PO). Ethanolic extract did not exhibit any significant activity. CONCLUSIONS: Petroleum ether extract and B hispida seed oil inhibited testosterone-induced hyperplasia of the prostate in these rats. Further studies are needed to evaluate its effect in humans with benign prostatic hyperplasia.

An update on Shankhpushpi, a cognition-boosting Ayurvedic medicine.

Shankhpushpi is an Ayurvedic drug used for its action on the central nervous system, especially for boosting memory and improving intellect. Quantum of information gained from Ayurvedic and other Sanskrit literature revealed the existence of four different plant species under the name of Shankhpushpi, which is used in various Ayurvedic prescriptions described in ancient texts, singly or in combination with other herbs. The sources comprise of entire herbs with following botanicals viz., Convulvulus pluricaulis Choisy. (Convulvulaceae), Evolvulus alsinoides Linn. (Convulvulaceae), Clitoria ternatea Linn. (Papilionaceae) and Canscora decussata Schult. (Gentianaceae). A review on the available scientific information in terms of pharmacognostical characteristics, chemical constituents, pharmacological activities, preclinical and clinical applications of controversial sources of Shankhpushpi is prepared with a view to review scientific work undertaken on Shankhpushpi. It may provide parameters of differentiation and permit appreciation of variability of drug action by use of different botanical sources.

Effect of Convulvulus pluricaulis Choisy. on learning behaviour and memory enhancement activity in rodents.

The present study was undertaken to investigate the effects of Convulvulus pluricaulis (CP), considered as Shankhpushpi on learning and memory in rodents. Nootropic activity using Cook and Weidley's Pole Climbing Apparatus, passive avoidance paradigms and active avoidance tests were used to test learning and memory. The ethanolic extract of CP and its ethyl acetate and aqueous fractions were evaluated for their memory enhancing properties. Two doses (100 and 200 mg kg(-1) p.o.) of the ethanolic extract and ethyl acetate and aqueous fractions were administered in separate groups of animals. Both the doses of all the extracts of CP significantly improved learning and memory in rats. Furthermore, these doses significantly reversed the amnesia induced by scopolamine (0.3 mg kg(-1) i.p.). Nootropic activity was compared using piracetam as the standard. Moreover, CP has exhibited potent memory-enhancing effects in the step-down and shuttle-box avoidance paradigms. Further studies are necessitated to identify the exact mechanism of action.