Effects Of Soothing Liver, Invigorating Spleen Methods And Recipes Of TCM On Expression Of LXRα MRNA And Protein In Hepatic Tissue Of Rats With Non-alcoholic Fatty Liver Disease

Objective:To observe the effect of soothing liver and invigorating spleen recipes on LXRa mRNA and protein expression in hepatic tissue of rats with non-alcoholic fatty liver disease(NAFLD), and to study the possible mechanism on invigorating spleen, soothing liver methods and recipes resistance to the NAFLD, for further providing the experimental evidence on prevention and treatment of NAFLD.Methods:After one week adaptive feed,55 male SD rats were randomly divided into 5 groups: normal group, model group, integrated group(Chai hu su gan san and Sen lin bai zhu san), soothing liver group(Chai hu su gan san) and invigorating spleen group(Sen lin bai zhu san), the model group 15 rats and each of the other groups 10 rats. Rats in the normal group were fed with basic diet, and those in the remainder groups were fed with high-fat emulsion by gastric perfusion(10mL/kg-BW) besides basic diet. Meanwhile, Rats in the intervention group were administered with the correspondence intervention recipe; rats in the normal and model groups were administered with normal saline. At the end of sixth and eighth week, two rats were randomly selected from model group to observe hepatic histopathology. The rat model of NAFLD was successfully confirmed by hematoxylin and eosin staining(HE) at the end of eighth week. After 12 hours by fasting but drinking water freely, all rats were anesthetized by 3% pentobarbital sodium through intraperitoneal injection. Blood samples were collected from the aortaventralis. The content of TC, TG, ALT, AST in the serum, as well as TC, TG in the hepatic homogenate were detected by automatic biochemical analyzer. The histopathological changes of hepatic tissue were observed by HE staining. The expression levels of LXRa mRNA in liver tissue were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The expression of protein LXRa in the hepatic tissue were examined by immunohistochemistry.Results:1. Biochemical analysis:compared with normal group, the serum ALT and AST levels in model group were significantly increased (P<0.05,P<0.01); the serum TC and TG contents in model group were dramatically increased(P<0.05); the liver levels of TC and TG in model group were significantly increased(P<0.01,P<0.05). compared with the model group, the level of serum ALT in each treatment group was lower but there was no statistics differences between them(P>0.05); the level of serum AST in each treatment group was significantly decreased(P<0.01); the serum TC and TG contents in each treatment group were significantly decreased(P<0.01,P<0.05); the liver content of TC in each treatment group, the liver content of TG in soothing liver group and invigorating spleen group were were significantly decreased(P<0.05); but the liver content of TG in integrated group was lower but no statistics differences.2. Light microscope observation:Light microscope observation revealed no obvious lesions in normal group:hepatic tissue histological structure integrated, hepatic lobules clear hepatocytes cord-like permutation orderly. There were revealed medium fatty degeneration in hepatocytes in model group:swelling of hepatoeytes and different sizes of lipid droplets inside the cytoplasm, inflammatory cells were seen in the portal area. But the pathological changes in each treatment group were ameliorated in different degree, that in the soothing liver group was eased relativly while the integrated group were worst.3. The expression of LXRa mRNA and protein:In each group, the LXRαof mRNA and protein in the hepatic tissue were expressed in different degree, it was the lowest in normal group. Compared with the normal group, LXRαwere higher in model group(P<0.05). Compared with the model group, the expression of LXRαmRNA and protein in each treatment group were lower(P<0.05 or P<0.01), especially in the invigorating spleen group.Conclusion:1. Rats model of NAFLD were successfully duplicated by high-fat emulsion gastric perfusion for 8 weeks.2. High fat diet is a major cause of NAFLD, while liver and spleen dysfunction may be the main pathogenesis.3. The high expression of LXRαmRNA and protein may be an important mechanism for the formation of NAFLD.4. The soothing liver and invigorating spleen recipes can improve hepatocytes steatosis in rats with NAFLD. The mechanism may be through down-regulating expression of the liver LXRαmRNA and protein, regulation of lipid metabolism, protect the liver cells and reduce liver lipid accumulation. LXRαmay an important target for soothing liver and invigorating spleen recipes resistance to the NAFLD.