The Research Of Protective Effect And Possible Mechanisms Of Modified YinChenWuLingSan Agains Experimental Hepatic Injury In Mice

Objective:The research was According to Traditional Chinese medicine theory, based on the existing discovery of viral hepatitis,use D-GalN-induced chemical hepatic injury model in rat and BCG+LPS-induced immunological hepatic injury model in mice as the research object,to study the effects and the control mechanism of ModifiedYinChenWuLingSan on experimental liver injury in animals.To provide experimental basis for ModifiedYinChenWuLingSan for the treatment of viral hepatitis,and also lay the foundation for this rescriptions further study.Methods:1.The study of protective effect of ModifiedYinChenWuLingSan against chemical hepatic iniury in rat:Randomly divided 60 SD-rat into normal control group,model group,Positive control group,ModifiedYinChenWuLingSan low dose, medium dose,and high dose groups.Treatment group and positive control group began to intragastric administration,at the same time,to supply the nomal group and model group with normal saline in the same volume.It last 10 days.In addition to the normal control group,The remaining rats were disposable intraperitoneal injected D-GaIN at the dose of 400mg/kg at the 9th day after the beginning of the experiment,Normal group intraperitoneal injection the same dose of normal saline.Collected blood and liver tissues after 24 hours, The serum activity of ALT,AST were analyzed,the serum level of T-Bil and serum NO were measured,The content of MDA and the activity of GSH-Px in liver homogenate were measured.Dimensions of hepatic damage in the histopathology were observed.2.The study of protective effect of ModifiedYinChenWuLingSan against immunological hepatic iniury in model mice:Randomly divided 90 NIH-mice into normal control group,BCG+LPS-Model group,Positive control group, ModifiedYinChenWuLingSan low dose,medium dose,and high dose groups.Mices in Each group were tail vein injected BCG liquor about 2.96×10⁷ viable bacteria at the first day of the experiment.Treatment group and positive control group start intragastric administrated from the first day,at the same time,to supply the nomal group and model group with normal saline in the same volume,lasted 10 days.Addition to the mices of the blank group tail vein injected saline at the dose of 0.2ml/20g,the mices of other groups all tail vein injected LPS at the dose of 7μg/20g.Collected blood after 10 hours,the serum activity of ALTandAST were analyzed,the content of MDA and the activity of SOD in liver homogenate were measured.The content of NO and TNF-a in serum were measured also.The liver and spleen were aseptic removed and weighed, organ(liver,spleen)index were measured,T,B lymphocyte proliferation were determined by MTT colorimetric assay,and dimensions of hepatic damage in the histopathology were observed.3.The study of influence of ModifiedYinChenWuLingSan on immune function: Randomly divided 60 NIH-mice into normal control group,model group, ModifiedYinChenWuLingSan low dose,medium dose,and high dose groups.Treatment groups were continuate intragastric administrated for 8 days.At the 20min after the first administration,mices of model group and treatment groups were injected hydrocortisone at the dose of 5mg/kg for 3 days,to supply the nomal group and model group with normal saline in the same volume.15min after the last administration tail vein injected 25%Chinese ink at dose of 0.1ml/10g. Separately collected blood 20μl after 2min and 20 min,the blood was dissolve in 0.1%Na₂CO₃ solution,CarbonExpurgatory index(K) and Phagocytosis index(a) were calculated.Removed liver,spleen and thymus,and weighed it, organ(liver,spleen,thymus)index were calculated.Results:1.The protective effect of ModifiedYinChenWuLingSan against chemical hepatic iniury in rat:The serum activity of ALT,AST and the serum level of T-Bil in chemical hepatic iniury rat model of medium dose group and high dose group were significantly reduced,and pathology hepatic damage of medium dose group and high dose group were observably relieved,the effect of the high doses drug was most obvious.ModifiedYinChenWuLingSan was also found to significantly decreased liver homogenate increased the MDA levels,improved liver homogenate GSH-Px enzyme activity,asignificant reduced serum NO content in chemical hepatic iniury models.2.Protective effect of ModifiedYinChenWuLingSan against immunological hepatic iniury in mice:medium dose drug and high dose drug of ModifiedYinChenWuLingSan was found to significantly decrease the serum ALT and AST activities of model mice by immunized liver damage,and ModifiedYinChenWuLingSan medium dosage drug have better effect than WuZhipill,Modified YinchenWulingSan was also found to drestrain the enlargerment of liver and spleen,reduce the putrescence of hepatic cell.Mean while significantly decreased MDA content and SOD activities in liver homogenate,reduce the concontent of NO and TNF-a in serum obviously,controled splenic T,B lymphocyte proliferation in BCB/LPS liver injury mice.3.Influence of ModifiedYinChenWuLingSan on immune function:Medium doses drug of ModifiedYinChenWuLingSan could enhance the macrophage phagocytic function of immunocompromised mice,and could significantly promote inhibited immune organs recovery and growth.Conclusion:ModifiedYinChenWuLingSan have significant protective effect to D-GaIN induced chemical liver injury in rats and BCG/LPS-induced immunological liver injury in mice.It was mainly manifested in the reduction of serum aminotransferase,bilirubin increased in liver injury animal,and the protection effect to liver tissue and cells.Analysed from the test results of liver tissue MDA levels,GSH-Px activity,SOD activity,serum TNF-a concentration,NO content,as well as splenic T,B lymphocyte proliferation in liver injury mice,it was likely to be related to the capability of resiste stress of oxidation,enhance activity of free radical scavenging enzyme,reduce the generation of inflammatory mediators,promote regeneration of liver cell, and it's fuction of immuneregulation.Besides,experimental results of the influence on immune function demonstrated that ModifiedYinChenWuLingSan can significantly improve its macrophage phagocytosis,enhance the nonspecific immune response.So it could be confirmed that ModifiedYinChenWuLingSan have the fuction of adjust the immunity system,the capbility of protect liver cells and agains hepatic injury was likely to be related to it's two-ways regulated fuction on immunity.