疏肝健脾方药对非酒精性脂肪性肝病大鼠肝组织LXRα mRNA及蛋白表达的影响

Effects of soothing liver,invigorating spleen recipes on expression of LXRα mR NA and protein in hepatic tissue of rats with non-alcoholic fatty liver disease

目的探讨疏肝健脾方药对非酒精性脂肪性肝病(NAFLD)大鼠肝组织LXRαmRNA及蛋白表达的影响。方法选用SD大鼠55只,随机分为正常组、模型组、疏肝组(灌服3.2 g.kg-1·d-1剂量的柴胡疏肝散)、健脾组(灌服10.0 g.kg-1·d-1剂量的参苓白术散)、综合组(灌服11.9 g.kg-1·d-1剂量的柴胡疏肝散和参苓白术散合方),模型组15只,其余各组10只。采用灌饲高脂肪乳剂(10 ml/kg)的方法复制大鼠NAFLD实验动物模型,给药8 w后处死动物,腹主动脉采血,用全自动生化分析仪检测血脂及肝功;常规HE染色观察肝组织病理变化;RT-PCR方法检测肝组织LXRαmRNA的表达;免疫组织化学方法检测肝组织LXRα蛋白的表达。结果与正常组相比,模型组大鼠肝细胞脂肪变性明显,血脂及肝功均有不同程度的升高(P<0.05,P<0.01),大鼠肝组织LXRαmRNA及蛋白表达明显升高(P<0.01);各给药组血脂及肝功和肝组织LXRαmR-NA及蛋白的表达均较模型组显著降低(P<0.05,P<0.01),其中以健脾组下降最为明显。结论疏肝健脾方药对高脂饮食诱导的大鼠NAFLD有较好的治疗作用,其机制可能与其下调肝脏LXRα的表达有关。

Objective To investigate the effects of soo thing liver and invigor ating spleen recipes on the expression of LXRα mRNA and protein in hepatic tiss ue of rats with non-alcoholic fatty liver disease（NAFLD）. Methods 55 male SD rats were randomly divided into 5 groups： normal,model,soothing liver（receivi ng gavage of Chaihu Shugan Powder 3.2 g·kg-1·d-1）,invigorating s pleen（rece iving gavage of Shen Ling Baizhu Powder 10.0 g·kg-1·d-1） and inte grated grou ps（receiving gavage of Chaihu Shugan Powder and Shen Ling Baizhu Powder combina tion recipes11.9 g·kg-1·d-1）.15 rats were in model group and 10 rats were i n each of the other groups.Except the normal group,the rats in other groups w e re fed with high-fat emulsion by gastric perfusion（10 ml/kg） besides basic die t to induce NAFLD.After treatment for 8 weeks,the rats were executed to obtain blood samples from aortaventralis.The contents of TC,TG,ALT,AST in the seru m,as well as TC,TG in the hepatic homogenate were detected by automatic biochem ical analyzer.The histopathological changes of hepatic tissue were observed by HE staining.The expression levels of LXRα mRNA in liver tissue were analyzed b y reverse transcription polymerase chain reaction（RT-PCR）.The expression of protein LXRα in the hepatic tissue was examined by immunohistochemistry. Results Compared with normal group,there were revealed m edium fatty degeneration in h epatocytes in model group;the serum TC,TG,ALT,AST levels and the liver level s of TC and TG in model group were increased in different degree（P0.05, P0.01）;the expression of LXRα mRNA and protein in the hepatic tissue were dramatica lly increased（P0.01）.Compared with the normal group,the serum TC,TG, ALT,A ST levels and the liver levels of TC,TG and the expression of LXRα mRNA and pr otein in each treatment group were decreased in different degree（P0.05, P0.01）,especially in the invigorating spleen group. Conclusions Soothing liver and i nvigorating spleen methods and recipes show certain therapeutic effect on rats w ith NAFLD,which may be due to down-regulating expression of the liver LXRα mR NA and protein in hepatocyte.