Protective Effect Of Acanthopanax Senticosus Polysacharides On The Mice With Immunological Liver Injury

ObjectiveUsing metabonomics method of Acanthopanax Senticosus Polysaccharides on mice with immunological liver injury protective effect,looking for potential biological markers and analysis of the metabolic pathway,determine Acanthopanax Senticosus Polysaccharides in play a role in the group and mechanism of action.Methods1.Forty BALB/c mice were randomly assigned to four groups,the control group,Acanthopanax Senticosus Polysaccharides treatment group(physiological medicine group),concanavalin A(ConA)model group and ConA model drug delivery group(pathological treatment group),the control group and the model group give physiological saline(10mg/mL),physiological medicine group and pathological treatment group give Acanthopanax Senticosus Polysaccharides solution 14.5mg/mL continuous irrigation stomach for 7 days.In addition to the control group and the physiological medicine group,the other two groups were injected with ConA solution(2.0mg/mL)at the end of the seventh day after administration of 0.5h,and the liver and spleen were taken from the 8h after fasting.Preparation of 10%liver and spleen tissue homogenate.Using metabonomics method of mouse liver and spleen tissue homogenate analysis of physiological and pathological conditions,to find the potential biological markers and metabolic pathways.2.The D941 resin was used to remove protein and remove the pigment from the Acanthopanax Senticosus Polysaccharides solution of 10mg/mL.DEAE-Sepherose F.F anion exchange resin column and Sephadex-G200 molecular sieve column are further separated and purified to remove the protein and pigment.The Coomassie brilliant blue method removal rate of protein were measured,by phenol-sulfuric acid method was used to determine the content of polysaccharide,and measured the OD value of wavelength 420nm,calculate pigment removal rate.3.Sixty BALB/c mice were randomly assigned to six groups:control group,concanavalin A(ConA)model group,positive drug(biphenyl diester)group and ASPS-1-1 group,ASPS-2-1 group,ASPS-3-1 group,control group and model group were given 0.9%saline(lOmg/mL)and positive drug group(20mg/mL),ASPS-1-1 group(6.9mg/mL),ASPS-2-1 group(4.6mg/mL),ASPS-3-1 group(2.0mg/mL)intragastric administration,for 7 consecutive days,immune liver injury model was induced by ConA,take mice serum,liver and spleen of the standby.Liver homogenate was used to detect the levels of GSH,SOD,MDA,NO,IL-1? and TNF-a.Serum and spleen tissue homogenate were used to detect IL-2,IL-4,IL-6 and IFN-y levels.Results1.Under the physiological state in the liver of mice selected 27 biomarkers,which is ion mode under 19,negative ion mode under 8,spleen screened 53 biomarkers is ion mode under 21,negative ion mode under 32,related metabolic pathway have sphingolipid metabolism,purine metabolism,glutathione metabolism and bile secretion.In the pathological state in the liver of mice selected 36 biomarkers,which is ion mode under 17,negative ion mode under 19,spleen screened 41 biomarkers is ion mode under 24,negative ion mode under 17,related to the metabolic pathway have cysteine and methionine metabolism,lemon acid cycle,purine metabolism and so on.2.After the initial removal of the D941 resin column,the removal rate of the protein was 88.26%,the decolorization rate was 88.66%,the retention rate of polysaccharide was 82.81%.The resulting solution is treated by DEAE-Sepherose F.F,three single symmetrical peaks can be obtained,shows that Acanthopanax senticosus polysaccharides were divided into three relatively pure group points,was named the ASPS-1,ASPS-2 and ASPS-3.After further purification,the Sephadex-G200 gel filtration column was purified to obtain 1 single symmetrical peaks,which indicated that the purified polysaccharide was purified and the 3 fractions were named ASPS-1-1,ASPS-2-1 and ASPS-3-1.The percentages of ASPS-1,ASPS-2 and ASPS-3 were 47%,32%and 13%respectively,and the ASPS-1 was the neutral polysaccharide,and the ASPS-2 and ASPS-3 groups were divided into acidic polysaccharides.3.Compared with the model group,the ASPS-1-1 can reduce the liver index and spleen index of mice;increased in liver tissue GSH and SOD level,reduce MDA,IL-1,NO and TNF-a level;reduce the spleen tissue and serum IL-2,IL-4,IL-6 and IFN-y level.ASPS-2-1 can reduce the liver index;increase the level of GSH in liver tissue,reduce the level of serum of IL-2 and TNF-a,MDA in liver tissueConclusion1.Pathological and physiological state under the condition of mice liver and spleen tissues in a total of 23 common biomarkers,polysaccharide of Acanthopanax senticosus Polysaccharides mainly through the physiological and pathological state intervention is some glutathione metabolism,purine metabolism,cysteine and methionine metabolism pathway play a role.In addition,there are non common of citric acid cycle,bile secretion,taurine metabolism and other pathways in the liver damage also play a role.It is possible to protect the immune liver injury mice by improving the immune function,anti inflammation,anti oxidation,and the supply of energy and so on.2.Acanthopanax Senticosus Polysaccharides by D941 resin column preliminary impurity removal,and by DEAE-Sepherose F.F column for separation and purification of neutral polysaccharide ASPS-1,acidic polysaccharide ASPS-2 and ASPS-3 three groups,which ASPS-1 in Acanthopanax Senticosus Polysaccharides which accounted for proportion most,after Sephadex-G200 gel filtration column further separation purified ASPS-1-1,ASPS-2-1 and ASPS-3-1 three homogeneous group.3.ASPS-1-1 component in the protection and restoration of immunological liver injury plays main role,mainly through the regulation of inflammatory cytokine levels and oxidative stress related enzymes activity to achieve its protective effect on immunological liver injury.In addition,the ASPS-2-1 component also has a protective effect on liver injury.