Study On Chemical Constituents And Hepato Protective Effect Of Schisandra Chinensis Polysaccharides Before And After Steaming With Rice Vinegar

Schisandra chinensis was contained in Shen Nong’s herbal classic at first.It is the desiccative ripe fruit of Schisandra chinensis(Turcz.)Baill.with sour and sweet in taste,warm in nature,which belongs to the lung,heart and kidney.The main effects of Schisandra chinensis are astringent,enhancing Qi while nourishing fluid,tonifying kidney,and calming heart,etc.Schisandra chinensis has the long history of medicinal uses.Because of its extremely high medicinal,it was highly praised by doctors in past dynasties.And it recorded that“If people use it as the Tonic medicine,it must be used in the north.”Schisandra chinensis mainly contains the lignans,the volatile oils,the triterpenes,the flavonoids,the organic acids,and polysaccharides,etc.But,all the time,the researches of Schisandra chinensis restrain the lignans and the triterpenes.And there are seldom researchers study the chemical components and pharmacological activities of Schisandra chinensis polysaccharides before and after steaming with rice vinegar.Schisandra chinensis is full of the polysaccharides,the total polysaccharides account for 7-9%.This thesis mainly studies on the variations of Schisandra chinensis polysaccharides before and after steaming with rice vinegar about their polysaccharides’extractions,components and hepato protective effect at the first time.In order to provide references for the reasonable development and utilization of Schisandra chinensis polysaccharides before and after steaming with rice vinegar.Experimental methods:1.In accordance with the fourth part of the Chinese Pharmacopoeia(2015 edition)general rules,to get the vinegar steamed Schisandra chinensis.Extractions of Schisandra chinensis polysaccharides with the classical technique of alcohol sedimentation.Then measured the total sugar contents with phenol-sulfuric acid method.Measured the uronic acid contents with the carbazole-sulfuric acid method.Measured the protein content with Coomassie brilliant blue staining method.2.The total Schisandra chinensis polysaccharides before and after steaming with rice vinegar were separated and purified by various ion exchange and gel-filtration chromatography,which including FPA90-Cl~-combine FPC-3500,DEAE Cellulose-52 and DEAE-Sepharose F.F.The polysaccharides components were all analyzed by HPLC to identify their purities.3.HPLC detected average molecular weight of each pure homogeneous polysaccharide.Labeled by PMP derivatization and analyzed by HPLC to detect monosaccharide compositions and molar ratios of each pure homogeneous polysaccharide,which from the total Schisandra chinensis polysaccharides before and after steaming with vinegar.4.This thesis choosed the mice model with auto immune hepatitis,which accused by LPS combination D-GalN via abdominal.Mice were orally administered with aqueous solutions(400 mg/kg)of crude Schisandra chinensis polysaccharides and vinegar steamed Schisandra chinensis polysaccharides for pre-protection treatment.Calculated the various of liver index,spleen index,and thymus index in differ groups mice.Observed the pathological changes of liver tissue in mice with the Hematoxylin-eosin HE staining method.Detected ALT and AST levels of mice’s liver homogenate with the Microplate method.Detected TNF-αand IL-1βlevels of mice’s serum with the Enzyme-linked immunosorbent assay method.Experimental results:1.The yields of Schisandra chinensis total polysaccharides before and after steaming with rice vinegar were 9.35%and 7.24%respectively of the plant raw material.Crude Schisandra chinensis total polysaccharides is brown flocculent solid,and Schisandra chinensis total polysaccharides steamed with rice vinegar is dark brown flocculent solid.Both of them are easily soluble in water,highly soluble in hot water,and insoluble in organic solvents.Detected the total sugar contents of Schisandra chinensis before and after steaming with rice vinegar were 79.36%and 76.74%respectively.The urnic acid contents of them were 36.14%and 38.57%respectively.The protein contents of them were 6.98%and 7.02%respectively.2.Each purified homogeneous polysaccharide of Schisandra chinensis total polysaccharides before and after steaming with rice vinegar:the total polysaccharides of crude Schisandra chinensis and steamed with rice vinegar were eluted by Amberlite FPA90-Cl~-combine Amberlite FPC-3500 anion series resin.The eluted components were all heteropolysaccharides,which detected by HPLC.And four purified polysaccharide components were obtained by separation and purification of DEAE Cellulose-52 and DEAE-Sepharose F.F ion exchange agarose gel:SP-1 and SP-4 which are in crude Schisandra chinensis,VSP-1 and VSP-2 which are in rice vinegar steamed Schisandra chinensis.The state of them are light yellow powder and light brown,soluble in water,highly soluble in hot water,insoluble in organic solvents.3.Preliminary analysis of chemical composition of homogeneous polysaccharides from Schisandra chinensis before and after steaming with rice vinegar.The average molecular weight of SP-1 is 1.06×10~9 Da,and it is composed by Rha:GalUA:Glc:Gal:Ara=5.94:11.25:19.10:13.49:4.93.Average molecular weight of SP-4 is 6.05×10~8 Da,and it is composed by Man:Rha:GalUA:Glc:Ara=12.68:20.41:0.71:36.79:3.51.The average molecular weight of VSP-1 is 2.17×10~8 Da,and it is composed by Rha:GalUA:Glc:Xyl=14.92:9.75:38.45:4.61.The average molecular weight of VSP-2 is 1.70×10~8 Da,and it is composed by Man:Rha:GalUA:Glc:Ara=34.27:1.82:15.56:142.33:62.94.4.It was known from the results of calculations and detections that the total Schisandra chinensis polysaccharides after steaming with rice vinegar could decline the liver index,spleen index,thymus index,ALT and AST levels,and TNF-αand IL-1βlevels.The Schisandra chinensis polysaccharides after steaming with rice vinegar also could improve the pathological changes of liver tissue in mice.The hepato protective effect in vinegar steamed polysaccharides was better than that in crude polysaccharides.Experimental conclusion:This thesis preliminary studies on the polysaccharides of Schisandra chinensis before and after steaming with rice vinegar.By comparing of the polysaccharides of Schisandra chinensis before and after steaming with rice vinegar,it proved the total polysaccharide of Schisandra chinensis steamed with rice vinegar was differ from the crude Schisandra chinensis in color and chemical component.By comparing the hepato protective effect of the polysaccharides of Schisandra chinensis before and after steaming with rice vinegar,it proved the hepota protective activities are not only the lignans,but also the polysaccharides.And the polysaccharides are also the material basis of hepato protective effect.Else,the steaming with the rice vinegar could enhance the hepato protective effect of crude Schisandra chinensis polysaccharides.They all provide the references for the chemical structure analysis,the mechanism of hepato protective effect,and reasonable development and utilization of Schisandra chinensis polysaccharides before and after steaming with rice vinegar.