Study On The Mechanism Of "Wenyang Jiedu Huayu" Prescription In Acute-on-Chronic Liver Failure Based On Immune Steady State In Intestinal Mucosa

Objective:The intervention mechanism of“WenYang JieDu HuaYu Prescription”on ACLF rat is studied from the histopathology of liver;intestine mucosa;ultrastructural changes of intestinal mucosal immune barrier.Methods:1.ACLF rat model was established by D-Gal+LPS on the fibrosis rat.2.The 40 ACLF rats were randomly divided into:M Group,Z_lowow group,Z_highigh group,C group,each group of 10,in addition N group were selected.fresh feces was collected and detected by 16S rRNA technique.TBil,ALT and AST was analyzered.Detection of endotoxin by serum tachypleus amebocyte lysate;TNF-?,IL-6 and IL-1 levels were determined by ELISA;histopathology of liver tissue and small intestinal mucosa was determined by HE and transmission electron microscopy,The expression of TLR4?occludin?ZO-1 were observed by PCR and Western-Blot technique.Results:1.HE staining of rat liver tissues showed that The area of hepatocyte necrosis was smaller in Z_highigh group.2.The HE staining of small intestine mucosa showed,the intestinal mucosa was the least damaged,and the mucosal surface was smooth without obvious shedding in the Z_highigh group.3.by 16S rRNA technology,Using Shannon index,Observed OTU index,Faith's index and Chao1 method,the M group were significantly lower than the Z_highigh and C group.Analysis by Alpha and Beta method,There were significant differences between the Z_high?M and Z_lowow group?P<0.01?;between C?M and Z_lowow group?P<0.05?;There was no significant difference between C and Z_highigh group?P>0.05?.4.Comparison of ALT,AST and TBil values in peripheral blood:These index decreased more effectively in Z_highigh group.The efficacy of Z_highigh and Z_lowow group in reducing TBil was similar?P>0.05?;5.Comparison of serum TNF-?,IL-6,and IL-1,All the experimental groups were significantly lower than the M group?P<0.05?.the Z_highigh group was more significant reducted?P<0.05?.the effect of TNF-?decreased more significantly in the Z_highigh and the C group?P<0.05?,and the effect of TNF-?decreased equally in the Z_highigh and C group?P>0.05?;the LPS of the Z_highigh group was significantly reduced?P<0.05?;6.Comparison of TLR4mRNA in liver tissues,zo-1mrna,occuldin mRNA in small intestine expression,the TLR4 decreased and the occuldin mRNA expression increased more significantly in Z_highigh and C group?P<0.05?,while the Z_highigh group showed no difference from the C group?P>0.05?.Conclusion:1.“WenYan gJieDu HuaYu Prescription”can significantly improve the abundance and diversity of intestinal flora and regulation of intestinal mucosal permeability,It can through Regulation of intestinal mucosal permeability and reduction of intestinal endotoxemia and inflammatory reaction,to reconstruct the intestinal mucosal immune homeostasis of ACLF rats,so as to inhibit the inflammatory necrosis of liver cells and reduce the mortality of ACLE.