Mechanisms Of Action Of "liver Disease And Spleen" From The Changes Of Motilin Signaling Pathway And Intestinal Flora In Rats With Hepatic Fibrosis

Objective:Establishing rat model of immunological hepatic fibrosis, we observe the variability of serum transaminase, total protein, albumin, motilin, motilin receptor in the gastric antrum and jejunum smooth muscle cells, intracellular free Ca2+concentration, portal endotoxin, ERIC-PCR, toll like receptor four expression levels, in order to clarify the mechanism of traditional Chinese medicine theory for liver-disorder involving spleen in the course of chronic liver disease. Comparative study of Xiaoyao Powder and its removing Chinese herbs of invigorating spleen investigate the mechanism of Chinese medicine therapy for treating liver by nourishing spleen on hepatic fibrosis.Methods:70Wistar rats, half male and female, were randomly divided into4groups: Blank control group(normal group), model group, control group, Xiaoyao Powder group(experimental group), normal group of10rats, the rest each group of20rats. The rat model of immunological hepatic fibrosis was established by injection of bovine serum albumin into tail vein for eight weeks. The normal group injected with the0.9%salt solution injection. As the same time, the experimental group was treated by intragastricing Xiaoyao Powder. Xiaoyao Powder removing atractylodes, poria, ginger and licorice was given to the control group as contrast, and were replaced by intragastricing sodium chloride in normal and model group, until the rat died. The rats were put to death and gather specimen24hours later after the end of the tail vein injection. Then we count and compare the serum levels of liver function, the pathological change degree of liver tissue, the level of serum MTL, the expression level of MTLR in the gastric antrum and jejunum smooth muscle cells,[Ca2+]i, the level of portal endotoxin, ERIC-PCR, the expression level of TLR4in liver tissue. Results:1. Liver function:Compared with normal group, the ALT\AST of rats in model group was increased, the TP and ALB was decreased, the differences were statistically significant (P<0.01). Compared with model group, the ALT(P<0.01)、AST(P<0.05) of rats in experimental group was decreased, the TP(P<0.01) was increased, the differences were statistically significant. There were no statistically differences in the ALB(P>0.05). The control group compared with the model group, no differences between the above indexes (P>0.05). The differences between experimental and control group were not statistically significant (P>0.05).2. Results of liver tissue pathology:Compared with normal group, the degree of liver fibrosis and fibrosis%were significantly increased in other group, and the difference was significant (P<0.01). Compared with model group, the degree of liver fibrosis and fibrosis%were decreased in experimental and control group, the differences were significant (P<0.05, P<0.01), but no significant difference between experimental and control group(p>0.05).3.MTL and its downstream signal:Compared with normal group, the level of serum MTL, the expression level of MTLR in the gastric antrum and jejunum smooth muscle cells,[Ca2+were decreased in model group, and the difference was significant (P<0.01). Compared with model group, the above indexes were increased in experimental group(P<0.05, P<0.01), and the MTLR in jejunum smooth muscle cells,[Ca2+]i of gastric antrum smooth muscle were increased in control group(P<0.01), but the difference was not significant (P>0.05) in the level of serum MTL, MTLR in the gastric antrum smooth muscle cells,[Ca2+]i of jejunum smooth muscle. The above indexes in experimental were higer than control group, except for MTLR in gastric antrum smooth muscle cells(P<0.05), the differences between experimental and control group were not significant (P>0.05).4. ERIC-PCR:Compared with normal group, the microbiota characteristics changed in depression model group, while experimental and control group returned it to normal at a great degree. Experimental group was better than control group. 5. Endotoxin:Compared with nomal group, endotoxin in portal vein was higher in model group, the differences was statistically significant (P<0.01). The endotoxin in experimental group was lower than model group, the differences were significant (P<0.05), but there was not significant between model and control group(P>0.05). The endotoxin in experimental was lower than control group, but there was not significant(P>0.05).6. Expression of TLR4:Compared with nomal group, expression of TLR4was higher in model and control group, the differences was statistically significant (P<0.01). Compared with model and control group, expression of TLR4were lower in experimental group, the differences was significant (P<0.01, P<0.05), but the differences between model and control group were not significant (P>0.05).Conclusion:1.The rat model of immunological hepatic fibrosis was decreased MTL and its downstream signal to weaken gastrointestinal motility, then changed the microbiota characteristics, increased portal endotoxin and TLR4. It is one of the mechanism that liver-disorder involved spleen in the course of chronic liver disease.2. Xiaoyao Powder could return the microbiota characteristics, decrease portal endotoxin and TLR4by increasing MTL and its downstream signal on hepatic fibrosis. It would be the important mechanism of Chinese medicine therapy for treating liver by nourishing spleen.