柴胡疏肝散合参苓白术散对非酒精性脂肪性肝病大鼠肝组织LXRαmRNA及蛋白表达的影响

Influence of Soothing Liver and Invigorating Spleen Recipes on LXR Alpha mRNA of Liver Tissue and Protein Expression of NAFLD Rats

目的:探讨疏肝健脾方药对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝组织LXRαmRNA及蛋白表达水平的影响。方法:选用SD大鼠55只,随机分为正常组10只、模型组15只、疏肝组10只、健脾组10只、综合组10只。正常组以基础饲料喂养,其余各组除基础饲料外,每天上午8∶00灌饲高脂肪乳剂10 mL.kg-1。所有动物自由进食饲料和饮清水,连续8周。各药物干预组大鼠在施以造模因素的同时,按体质量10 mL.kg-1给予相应药物,给药剂量按人和动物体表面积折算,其中疏肝组灌服柴胡疏肝散(含生药量0.32 kg.L-1),健脾组灌服参苓白术散(含生药量1.00 kg.L-1),综合组灌服柴胡疏肝散和参苓白术散的合方(含生药量1.19 kg.L-1),正常组和模型组灌服同样剂量的蒸馏水,每天下午1次。给药8周后处死动物,腹主动脉采血,用全自动生化分析仪检测血脂及肝功能;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蛋白的表达水平。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及蛋白表达水平明显升高(P<0.01);各给药组血脂、肝功能和肝组织LXRαmRNA及蛋白的表达水平均较模型组显著降低(P<0.05或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的表达水平有关。

Objective：To study the influence of soothing liver and invigorating spleen recipes on LXR alpha mRNA of liver tissue and protein expression of NAFLD rats.Methods：55 SD rats were randomly divided into the five groups,namely,normal group of 10 rats,model group of 15 rats,soothing liver group of 10 rats,invigorating spleen group of 10 rats,comprehensive group of 10 rats.The normal group was given basal feed,and other groups were given intragastric administrated with high fat emulsion 10 mL·kg-1 one time at 8：00 every morning.All rats took feed and drunk water at freedom for continuous for 8 weeks.Rats of drug intervention groups were given the corresponding drugs of 10 mL per kilogram weight,and the dosage of administration was discounted according to the human and animal body surface area.The soothing liver group was intragastric administrated with Caihu Shugan San（containing crude drug dose of 0.32 kg·L-1）,and the invigorating spleen group was intragastric administrated with Shenling Baizhu Power（containing crude drug dose of 1.00 kg·L-1）.the comprehensive group was intragastric administrated with Caihu Shugan San and Shenling Baizhu Power（Containing crude drug dose of 1.19 kg·L-1）.The normal group and the model group were given intragastric administrated with the same dose of distilled water,1 time every afternoon.The animals were sacrificed at 8weeks after medication.abdominal aortic blood,and blood lipid and liver function were tested by fully automatic biochemical analyzer.Conventional HE staining was used to observe the pathological changes of hepatic tissue;RT-PCR method was used for detection the expression of liver tissue LXR alpha mRNA;Immunohistochemical method was applied to detect the expression level of liver tissue LXR alpha protein.Results：Compared with the normal group,hepatocyte steatosis of the model group rats was apparent,and blood lipid and liver function of the model group had different degrees of increase（P0.05,P0.01）,and rat liver tissue LXR alpha mRNA and protein expression level of the model group increased significantly（P0.01）;Blood lipid,liver function and liver tissue LXR alpha mRNA and protein expression levels in each medication administration group were significantly lower than that in the model group（P0.05 or P0.01）,and among which,the invigorating spleen group was the most obviously declined.Conclusion：Soothing liver and invigorating spleen recipes has good therapeutical effect,and its mechanism may be related to its downgrade the expression level of liver LXR alpha.