Screening Of Anti-AD Active Fraction Extracted From Nourishing Spleen Yin Recipe

Objective: Nourishing spleen yin recipe (Zibu Piyin Recipe, ZBPYR) isdeveloped by Professor Libin Zhan, which can improve the patients’ memory andintelligent capabilities. It had been shown that ZBPYR had notable effects onneuroprotection, and it could treat the Alzheimer’s disease animal models. Therefore,this study was aimed to observe anti-AD activity of ZBPYR, screen anti-AD activefraction extracted from ZBPYR. ZBPYR is also determined the content ofpolysaccharides.Methods：1.Different fraction extracted from ZBPYR: ZBPYR is extracted byreflux extraction, then concentrate by rotary vacuum pump, separated by macroporousresin column or by alcohol precipitation at last.2.Establish Alzheimer’s disease animal models: Using intraperitoneal injection ofScopolamine hydrobromide to the mouse. And we observe the memory and intelligentcapabilities of the mouse through heliophobic test, step down test and Morris watermaze test. At last, we got the anti-AD active fraction extracted from ZBPYR.3.Experimental animal and Administration: Male Kunming mouse weighing20±2g were used. After3d adaptive feed, the mouse were randomly divided into9group, n=7. Each treatment group was given correspongding drugs, quantity ofadministration:0.4637g herbal medicine/10g.Control group and Alzheimer’s diseaseanimal models group were given the same amount of saline.4.The determination the content of polysaccharides in ZBPYR: Thepolysaccharides are determined by colourimetry with sulfuric acid-anthrone ascolourimetric reagent.Results:1.Evaluation of Alzheimer’s disease animal models: Behavioral resultsshowed that Alzheimer’s disease animal model mice’s learning and memory declined (P<0.01), compared with the control group.2.Evaluation of efficacy: The latency for the treatment groups of the alcoholsinking property and95%alcohol eluated to heliophobic test and step down theplatform was significantly prolonged as compared to that of the the Alzheimer’s diseaseanimal model mouse(P<0.01). The errors of the two groups were reduced (P<0.01). Inour Morris water maze tests, significantly shortened the escape latency time andswimming distance of the fraction of alcohol sinking property and95%alcohol eluate.During the peobe trial session, the alcohol sinking property and95%alcohol eluatedramatically improved the swimming time with the target quadrant(P<0.05).3.The results of the content of the polysaccharide: Sulfuric acid-anthrone waschosen as coloration reagent, the wave length was620nm. The regression equation wasy=1.566x+0.0654R~2=0.9948. The content of polysaccharide in ZBPYR was3.11%,precision: RSD=0.4%, reproducibility: RSD=1.7%RSD=3.44%, stability of samplesolution: RSD=1.81%.Conclusion:1.By using intraperitoneal injection of Scopolamine hydrobromidemethod we successfully established Alzheimer’s disease mouse model. Behavioralexperiments suggested that Alzheimer’s disease mouse exhibited learning and memorydysfunction.2.In the5fractions extracted from ZBPYR, the alcohol sinking property and95%alcohol eluate improved Alzheimer’s disease mice’s learning and memory capbility, itwas similar with ZBPYR. However, the other three fractions can not improvedAlzheimer’s disease mice’s learning and memory capability. Therefore, the alcoholsinking property and95%alcohol eluate may be the Anti-AD Active Fractions.3.The content of the polysaccharide is high in ZBPYR. Eatablished method wasconvient, reproductive and accurate to identify polysaccharides in ZBPY with theglucose as reference. These methods were simple, specific and repoductive.