Validation of ethnopharmacology of ayurvedic sarasvata ghrita and comparative evaluation of its neuroprotective effect with modern alcoholic and lipid based extracts in ß-amyloid induced memory impairment.

ETHNOPHARMACOLOGICAL RELEVANCE: Sarasvata ghrita (SG), a polyherbal formulation from ayurveda, an ancient medicinal system of India, has been used to improve intelligence and memory, treat speech delay, speaking difficulties and low digestion power in children. AIM OF THE STUDY: Study aimed to validate the ethno use of SG in memory enhancement through systematic scientific protocol. The effect of SG and modern extracts of ingredients of SG was compared on cognitive function and neuroprotection in amyloid-ß peptide 25-35(Aß25-35) induced memory impairment in wistar rats. Further the underlying mechanism for neuroprotective activity was investigated. MATERIALS AND METHODS: SG was prepared as per traditional method, ethanolic extract (EE) was prepared by conventional method and lipid based extract was prepared by modern extraction method. All extracts were standardised by newly developed HPLC method with respect to marker compounds. SG, EE and LE were administered orally to male Wistar rats at doses of 100,200 and 400 mg/kg Body Weight by feeding needle for a period of 21 days after the intracerebroventricular administration of Aß25-35 bilaterally. Spatial memory of rats was tested using Morris water maze (MWM) and Radial arm maze (RAM) test. The possible underlying mechanisms for the cognitive improvement exhibited by SG, EE and LE was investigated through ex-vivo brain antioxidant effect, monoamine level estimation, acetylcholine esterase (AchE) inhibitory effect and Brain-derived neurotropic factor (BDNF) levels estimation. RESULTS: SG, EE and LE were analyzed by HPLC method, results showed that EE extract has high percent of selected phytoconstituents as compared with SG and LE. SG and LE decrease escape latency and searching distance in a dose dependant manner during MWM test. In case of RAM significant decrease in number of errors and increase in number of correct choices indicate an elevation in retention and recall aspects of learning and memory after administration of SG an LE. SG and LE extract can efficiently prevent accumulation of ß-amyloid plaque in hippocampus region. There was increase in SOD, GSH, CAT and NO level and decrease in MDA levels in SG and LE administered animals. SG and LE have found to exhibit AchE inhibitiory activity and significant dose-dependant increase in BDNF level in the plasma. SG and LE significantly increased the levels of noradrenaline, dopamine and 5-hydroxytryptamine in the brain. CONCLUSION: The study validated the neuroprotective activity of SG. The study concludes the extraction efficiency of SG for selected phytoconstituents is less than modern methods. However the neuroprotective activity of SG and LE was found to be greater than EE.

Neuroprotective effect of Indian propolis in ß-amyloid induced memory deficit: Impact on behavioral and biochemical parameters in rats.

The study aimed at the investigation of neuroprotective activity of macerated ethanolic extract of Indian propolis (MEEP) against ß-Amyloid 25-35 (Aß25-35) induced memory impairment in Alzheimer's disease. MEEP was administrated orally to Wistar rats at doses of 100, 200 and 300mg/kg. Behavioral performances were evaluated using morris water maze and radial arm maze. At the end of behavioral study, the brains were removed and antioxidant parameters and brain monoamines were estimated. Further acetylcholinesterase (AchE) inhibition and brain-derived neurotropic factor (BDNF) were evaluated. In addition hematological parameters and histopathological tests were also carried out. In behavioral models, MEEP significantly (P<0.05) reversed the cognitive impairment of ß amyloid-induced rats. The antioxidant potential was significantly increased (P<0.05) after administration of MEEP. Malondialdehyde levels were significantly (P<0.01) decreased in brain homogenate after treatment with MEEP extract as compared with diseased control group (group III). MEEP showed dose-dependent AChE inhibition and increased the levels of brain monoamines (P<0.05) as compared with group III. MEEP improved memory deficits by increasing BDNF in plasma (P<0.05). The study concludes that MEEP has anti-Alzheimer potential in rats through multiple mechanisms and further studies are ongoing for fractionation and biological screening.

Marker-based standardization and investigation of nutraceutical potential of Indian propolis / 中西医结合学报

<p><b>OBJECTIVE</b>Propolis, a resinous material collected by honey bees from various plants, has been explored globally for its medicinal and nutritional properties. However, research over Indian propolis is at infancy. This study was designed to investigate nutraceutical potential of Indian propolis.</p><p><b>METHODS</b>In the present study, propolis extract was standardized with respect to markers caffeic acid phenethyl ester, caffeic acid, galangin, luteolin, curcumin, apigenin, pinocembrin and quercetin by new high-performance thin-layer chromatographic (HPTLC) methods. The physico-chemical analysis, residues analysis and in vitro antioxidant activity analysis were performed. Nutraceutical value was examined in terms of fats, fibers, minerals, proteins, polysaccharides, total carbohydrates, and energy value.</p><p><b>RESULTS</b>The developed HPTLC methods were found to be simple, reliable accurate, and the validation parameters were within the limits of the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use guidelines. Macerated ethanolic extract of propolis (MEEP) was found to have polyphenolic content of (20.99 ± 0.24) mg/g and flavonoids content of (8.39 ± 0.04) mg/g. MEEP was found to comprise of (283.33 ± 51.31) g/kg fats, (30.07 ± 7.30) g/kg fibers, (102.56 ± 2.84) g/kg proteins and (389.36 ± 57.50) g/kg carbohydrate with a calorie value of (38 409.33 ± 6 169.80) kJ/kg. It was found that Indian propolis exhibited high nutraceutical value and showed absence of pesticides and heavy metals. The MEEP showed in vitro antioxidant activity with inhibitory concentration of (12.24 ± 4.64) μg/mL.</p><p><b>CONCLUSION</b>The present work explores Indian propolis as a potential nutritious candidate. The proposed analytical methods can be applied in future screening of the quality of Indian propolis.</p>

HPTLC method for simultaneous analysis of stevioside and rebaudioside-A in Stevia rebaudiana.

An HPTLC method was developed and validated for simultaneous determination of stevioside (STE) and rebaudioside-A (REB-A) in Stevia rebaudiana leaves. The HPTLC separation was performed on precoated silica gel 60F254 HPTLC plates with the mobile phase ethyl acetate-ethanol-acetone-water (15 + 3 + 6 + 6, v/v/v/v). The densitometric quantification of steviol glycosides was carried out at Amax 580 nm in the reflection-absorption mode after spraying with anisaldehyde-sulfuric acid reagent. Rf values were 0.34 and 0.28 for STE and REB-A, respectively. The content of STE and REB-A in leaf extract was found to be 6.94 and 6.35%, respectively. The method was validated in terms of precision, accuracy, specificity, and robustness. The method can be useful for routine analysis of the sweeteners.