疏肝健脾方药对NAFLD大鼠肝组织PPARα mRNA和TNF-α蛋白表达的影响

Effects of Soothing Liver and Invigorating Spleen Recipes on PPARα mRNA and TNF-α Protein Expression in Non-alcoholic Fatty Liver Disease Model of Rats

目的:观察疏肝健脾方药对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠肝组织过氧化物酶体增殖物激活受体α(peroxisome proliferator activated receptor alpha,PPARα)mRNA和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)蛋白表达的影响及作用机制。方法:高脂饲料喂养大鼠12周建立NAFLD模型后,造模组大鼠随机分为模型组、疏肝组、健脾组、综合组、三七脂肝丸组和自然恢复组。除模型组继续高脂饮食外,其余各组均以基础饲料喂养,治疗组分别给予相应药物ig,其他各组给予相应蒸馏水ig。治疗8周后,腹主动脉采血,全自动生化仪检测血脂、肝功能及肝脂;常规HE染色观察肝组织病理变化;RT-PCR法检测肝组织PPARαmRNA表达;免疫组化法检测肝组织TNF-α蛋白表达。结果:与模型组比较,药物治疗各组及自然恢复组肝脏脂变程度明显减轻;血脂、肝脂、肝功能指数明显降低(P<0.01);肝组织PPARαmRNA表达均有不同程度的上调(P<0.05),以疏肝组、健脾组、综合组上调尤为明显(P<0.01);TNF-α阳性细胞表达明显减少(P<0.01),而以疏肝组、健脾组趋势最为显著。结论:疏肝健脾方药对高脂饮食诱导的NAFLD大鼠有较好的治疗作用,其机制可能与其上调PPARαmRNA和下调TNF-α蛋白表达水平有关。

Objective：To observe the effects of Soothing liver and Invigorating spleen recipes on PPARα mRNA and TNF-α protein expression in hepatic tissue of rats with non-alcoholic fatty liver disease（NAFLD） and to explore the molecular mechanism.Method：NAFLD model was established by feeding high fat diet for 12 weeks.Then rats were randomly divided into 6 groups： model group,Soothing liver group,Invigorating spleen group,integrated group,Sanqi Zhigan Pill group and natural recovery group.Rats in model group were continuously given high fat diet while all other rats were given normal diet.Meanwhile,rats in medication groups were administrated i.g.with corresponding herbs.Eight weeks after the treatment,all rats were anesthetized by 3% pentobarbital sodium through intraperitoneal injection.TC,TG,AST,ALT content in the serum and TC,TG content in liver tissue was detected by automatic biochemical analyzer.The pathological changes of hepatic tissue were observed by HE staining.The peroxisome proliferator activated receptor alpha（PPARα） mRNA expression in liver tissue was examined by reverse transcription polymerase chain reaction（RT-PCR）.tumor necrosrs factor-α（TNF-α） protein expression in the liver tissue was detected by immunohistochemistry.Result：Compared with the model group,the hepatic steatosis degree was alleviated obviously in all treatment groups and natural recovery group.TC,TG,AST,ALT content in serum,and TC,TG content in liver tissue were dramatically declined（P0.01）.PPARα mRNA expression in treatment groups and natural recovery group was up-regulated（P0.05）,especially in the soothing liver and invigorating spleen group,whose expression was much higher compared with model group（P0.01）;.Compared with that in model group,TNF-α positive cell numbers were significantly decreased in treatment groups and natural recovery group（P0.01）.Conclusions：Soothing liver and invigorating spleen methods and recipes showed certain therapeutic effect on NAFLD,which might be linked to up-regulating PPARα mRNA and down-regulating TNF-α protein expression.