Effect Of Soothing Liver And Invigorating Spleen Recipes On FAS MRNA And Protein Expression In Hepatic Tissue Of Rats With Non-alcoholic Fatty Liver Disease

Objective:To observe the effect of soothing liver and invigorating spleen recipes on FAS mRNA and protein expression in hepatic tissue of rats with non-alcoholic fatty liver disease (NAFLD),preliminary study the possible mechanism on invigorating spleen,soothing liver and their integrated methods and recipes resistance to the NAFLD. To povide experimental evidence for preventing and treating NAFLD.Methods:Fifty-five male SD rats were randomly divided into five groups:normal control group,model control group,invigorating spleen group, soothing liver group and integrated group.The normal control group were fed with basic forage, the rats in the rest of groups were administrated by gastric perfusion of formula intralipid.At the same time, the normal control group,model control group were administrated by gastric perfusion of normal saline, the rats in invigorating spleen group, soothing liver group, and integrated group were administrated by gastric perfusion of its group corresponding decoction.At the end of sixth,eighth week, two model rats were randomly selected for hepatic histopathology examination.The hematoxylin and eosin staining(HE) result confirmed the NAFLD modeling success at the end of eighth week. The rats were fastinged while give them enough water for twelve hours and then intraperitoneal injectioned with 3%sodium pentobarbital,and blooded sampling from the abdominal aorta, The serum ALT,AST,TC,TG contents and TC,TG contents in the hepatic homogenate were measured by automatic biochemical analyzer.HE method was used to observe the pathlology change of the rat hepatic tissue,the reverse transcription polymerase chain reaction technology was used to detect the expression of FAS of mRNA in the hepatic tissue of the rats, The protein of FAS expression in the experimental rats' hepatic tissue were examined by immunohistochemistry.Results:1. Compared with normal control group,the contents of serum ALT in the model group were higher (P<0.05);Compared with the model control group,the contents of serum ALT in the treating group were lower but there were no statistics differences between them(P>0.05);Compared with the normal control group,the contents of serum AST in the model control group were higher,there were statistics differences(P<0.01);While compared to model group,the contents of serum AST in the treating group were lower significantly(P<0.01).These changes showed that the model rats had some damages with hepatic cells and the soothing liver and invigorating spleen methods and recipes could protect the hepatoeytes.2. Compared with the normal control group, The contents of serum TC,TG in the model group were higher (P<0.05);The levels of serum TC,TG in the treating group were significantly lower than the model group(P<0.01,P<0.05);The contents of TC,TG in the hepatic homogenate were higher in the model control group(compared with the normal control group,P<0.01,P<0.05),the contents of TC in the treating group were lower than the model control group (P<0.05);while compared with model control group,the contents of TG in the invigorating spleen group,soothing liver group were lower(P<0.05). In the integrated group the contents of TG were low with no statistics differences(P>0.05). These showed that there were hyperlipemiathe in the model rats. and provided the information about the soothing liver and invigorating spleen methods and recipes could improve the liver lipid metabolism of the experimental rats.3. From the light microseope we could observe that the rats'degeneration of liver cells changes obviously in the model control group,swelling of hepatoeytes and different sizes of lipid droplets inside the cytoplasm.Inflammatory cells were seen in the portal area.But the pathology of the treated rats were improved in different degree, of which the soothing liver group eased relativly while the integrated group were worst.The result of pathology told us the modeling was success and the soothing liver and invigorating spleen methods and recipes could reduce the inflammatory response.4. The expression of FAS mRNA and protein in liver homogenate was low in the normal control group.Compared with the normal control group, the expression in the hepatic tissue of FAS mRNA and protein were higher in the model control group and had significant statistics differences(P<0.05); while compared with the model control group,the expression of FAS mRNA and protein in the treating groups were lower(P<0.05). the soothing liver group were the lowest but with no statistics differences(P>0.05). The high expression in the hepatic tissue of FAS mRNA and protein might be the one of importent mechanism which caused the NAFLD.The soothing liver and invigorating spleen methods and recipes might depress the expression of FAS mRNA and protein and then protect against steatosis injury of liver cells.The soothing liver method and recipe maybe had the better effect.Conclusion: 1. The rat model of NAFLD were successfully established by gavaged with formula intralipid.2. The soothing liver and invigorating spleen methods and recipes could extenuate the NAFLD rats' lipid metabolism and reducing serum AST and ALT,improve the rat lipid metabolism disorder,significantly reduce the hepatic damage of NAFLD.3. The expression in the hepatic tissue of FAS mRNA and protein were higher in the model rats'hepatic tissue, Soothing liver and invigorating spleen methods and recipes could decrease the expression of FAS mRNA and protein in the hepatic tissue,These suggested that the FAS might be one of the important target point to prevent NAFLD.