Study On The Hepato-protective Effect And Degradation Of Polysaccharide From Hyriopsis Cumingii (Lea)

Zhu Bang polysaccharide(ZBP) were extracted from the flesh of Hyriopsis cumingii(Lea).Which have the activity of hepato-protection in our early study.This paper focuses on the mechanism of hepato-protection in vitro.Different experiment methods were used to prepare the degradation of Zhu Bang polysaccharide(DZBP).The activities of hepato-protection of DZBP were studied.The main contents include four parts as follows:1.Literature studies.By referring to the literatures about ZhuBang,we know its historic evolution and modern research.By reviewing the literatures on animal polysaccharides,we know the characteristics and pharmacological activities.In addition,the literatures about the degradation methods of polysaccharides were consulted for experimental works.2.The study on the hepar-protective activity of the ZhuBang polysaccharide in vitroWe studied the mechanisms of ZhuBang polysaccharide,which has the protective effect on injured hepatocytes in vitro.The L-02 hepatocytes were incubated and then injured by CCl₄ and H₂O₂.The levels of AST,ALT,MDA and SOD in cultural supematant were detected by general methods.Cell viability were assayed by the method of MTT.The result showed that ZBP can reduce the levels of AST,ALT,MDA in cultural supematant which increased by CCl₄ and H₂O₂.It also can boost the hepatocytes viability and elevate the level of SOD which reduced by CCl₄ and H₂O₂.The results suggest that ZBP has direct protective activity on hepatocytes injured in vitro.It might be associated with the anti-oxidative activity of ZBP.3.Degradation of ZBP and effective fraction screeningThe molecular weight of ZBP was investigated by using Ubbleholde viscometry,the viscosity-average molecular weight was 4.64dL·g^-1.ZBP was resolved respectively byα-amylase,ultrasound and hydrogen dioxide in the study.We found that ZBP can be effectively degraded by using hydrogen dioxide.Using cell viability and oligosaccharide viscosity as the index of value,we adopted orthogonal experiment to investigate the condition of degradation.And we found the best condition:the concentration of H₂O₂ is 6%; the concentration of acetic acid is 3.5%;the reaction temperature is 50℃and the reaction time is 2 hours.The yield of DZBP was 60%.The viscosity-average molecular weight of DZBP was 1.74dL·g^-1.This method was simple and convenient.The L-02 hepatocytes were incubated and then injured by H₂O₂,the protective effects of DZBP were assayed by MTT.The CCl₄ and D-galactosamine Hydrochloride were used to injured the mice's liver.To compare the hepato-protection activities of ZBP and DZBP, giving by ig.or ip,we determined the levels of ALT,AST,SOD,MDA in serum.The results showed that the hepato-protection activity of DZBP was better,which were degraded by hydrogen dioxide.4.The semi-works production of Zhu Bang polysaccharideAccording to technology and operation in the early outcome,we researched on the semi-works production in KangYuan pharmaceutical llimited company,in order to conforming the need of industrialization and checking the rationality and feasibility of the technology.The major parameter of technology concluded that.The software of Hyriopsis cumingii(Lea) were cleaned,dripped and minced,and then boilled with 3 times of water after 30mins.The filter residue were boiled twice.We condensed the filter liquor which were combined.We added the ethanol to supermatant which were centrifugated with tube centrifuge,to make sure the ethanol concentration achieved 80%.Aafter overnight aging, we used vacuum filtration to obtain the precipitation,and dried the precipitation under reduced pressure at 60℃.The glyc- content of coarse polysaccharide must higher than 60%. The index was reached the government standard.We also instituted the standard of enterprise production.