疏肝健脾方药对NASH大鼠肝组织TLR4 mRNA及其蛋白表达的影响  

Effects of Soothing Liver and Invigorating Spleen Recipes on the mRNA and Protein Expression of TLR4 in Hepatic Tissues of Rats with NASH

作　　者：杨钦河[1] 黄进[1] 张玉佩[1] 王强[2] 何秀敏[1] 冯高飞[1] 王文晶[1] 李哲华[2] 

机构地区：[1]暨南大学,广东广州510632 [2]广州市番禺区中医院,广东广州511400

出　　处：《中药材》2013年第1期78-84,共7页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金资助项目(30973694);国家大学生创新性实验计划资助项目(091055930);广东省建设中医药强省立项资助课题(20111008);广州市番禺区科技计划资助项目(2011-Z-03-70)

摘　　要：目的:观察疏肝健脾方药对非酒精性脂肪性肝病(NASH)大鼠肝组织TLR4 mRNA及蛋白表达的影响,探讨疏肝健脾方药抗大鼠NASH的作用机制。方法:SPF级雄性SD大鼠72只,随机分为正常对照组、模型组、疏肝高剂量组(9.6 g/kg柴胡疏肝散)、疏肝低剂量组(3.2 g/kg柴胡疏肝散)、健脾高剂量组(30 g/kg参苓白术散)、健脾低剂量组(10 g/kg参苓白术散)、合方高剂量组(39.6 g/kg柴胡疏肝散合参苓白术散)、合方低剂量组(13.2 g/kg柴胡疏肝散合参苓白术散),每组9只。采用高脂饲料(10 mL/kg)喂养建立NASH大鼠模型,16 w后各组大鼠以3%戊巴比妥钠腹腔麻醉,腹主动脉采血,全自动生化仪检测血脂、肝脂和肝功能;HE和油红O染色观察肝组织病理变化;Real time Q-PCR检测肝组织TLR4 mRNA表达水平;Western blot检测肝组织TLR4蛋白表达水平。结果:与正常对照组比较,模型组大鼠肝细胞脂肪变性明显,肝细胞肿大,小叶内、汇管区炎细胞浸润,血清TC、LDL-C,肝组织TC、TG、肝组织TLR4 mRNA和蛋白表达均显著升高(P<0.01);与模型组比较,各用药组大鼠血脂、肝脂、肝组织TLR4 mRNA和蛋白表达均有不同程度降低(P<0.05或P<0.01),以高剂量作用最为显著。结论:疏肝健脾方药可能是通过下调TLR4 mRNA和蛋白表达水平发挥抗NASH的作用,TLR4可能是疏肝健脾方药抗NASH的有效靶点之一。疏肝健脾方药防治NASH可能存在一定的量效依赖关系。Objective：To research the effects of soothing liver and invigorating spleen recipes on expression of TLR4 mRNA and protein expression in hepatic tissue of rats with non-alcoholic steatohepatitis and its mechanism.Methods：72 male SD rats were randomly divided into 8 groups：normal group,model group,high-dose soothing liver group（receiving gavage of Chaihu Shugan Powder 9.6 g/kg）,low-dose soothing liver group（receiving gavage of Chaihu Shugan Powder 3.2 g/kg）,high-dose invigorating spleen group（receiving gavage of Shen Ling Baizhu Powder 30 g/kg）,low-dose invigorating spleen group（receiving gavage of Shen Ling Baizhu Powder 10 g/kg）,high-dose integrated Group（receiving gavage of Chaihu Shugan Powder and Shen Ling Baizhu Powder combination recipes 39.6 g/kg）,low-dose integrated Group（receiving gavage of Chaihu Shugan Powder and Shen Ling Baizhu Powder combination recipes 13.2 g/kg）,9 rats were in each group.Used high fat diet（10 mL/kg） to establish experiment model of NASH rat.At the end of the sixteenth weeks,the levels of serum lipids,liver lipids and serum aminotransferase were measured by automatic biochemical analyzer;Liver pathology was analyzed by HE and Oil red O staining;TLR4 mRNA was assayed by real-time fluorescent quantitative polymerase chain reaction（RT Q-PCR）;TLR4 protein was detected by Western blot.Results：Compared with normal group,the levels of TC,LDL-C in the serum,TC,TG as well as the expression of TLR4 mRNA and protein in the hepatic tissue were dramatically increased in model group（P0.01）.Compared with the model group,the levels of serum lipids,liver lipids,the expression of TLR4 mRNA and protein in the hepatic tissue were decreased in each treatment group（P0.05 or P0.01）.Conclusion：Soothing liver and invigorating spleen recipes can inhibit hepatic TLR4 expression,that may be one of their therapeutic mechanisms.There is much difference between high-does and low-does treatment groups in various testing items,which shows that there is does-effect

关 键 词：非酒精性脂肪性肝炎 疏肝健脾方药 TOLL样受体4 大鼠 

分 类 号：R285.5[医药卫生—中药学]