疏肝健脾方药对非酒精性脂肪性肝病大鼠肝组织UCP2 mRNA及蛋白表达的影响

Effect of Soothing Liver and Invigorating Spleen Recipes on UCP2 mRNA and Protein Expression in Hepatic Tissues of Rats with Non Alcoholic Fatty Liver Disease

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)大鼠肝组织解偶联蛋白2(uncoupling protein 2,UCP2)mRNA及蛋白表达的影响,初步探讨疏肝健脾方药治疗NAFLD的作用机制。方法采用高脂饲料喂养12周复制雄性SD大鼠NAFLD模型,将模型大鼠分为疏肝组、健脾组、疏肝健脾综合组(综合组)、三七脂肝丸组(三七组)、模型组,各组给予相应治疗8周。测定各组大鼠血清总胆固醇、三酰甘油、低密度脂蛋白胆固醇和高密度脂蛋白胆固醇含量。采用RT-PCR方法检测肝组织UCP2 mRNA的表达,免疫组织化学方法检测肝组织中UCP2蛋白活性变化。结果与正常组比较,模型组大鼠肝组织中UCP2 mRNA表达水平显著升高(P<0.05);与模型组比较,疏肝组、健脾组和综合组UCP2 mRNA表达水平显著下降(P<0.05)。模型组肝组织UCP2蛋白表达水平显著升高;与模型组比较,各用药组UCP2蛋白表达水平均显著下调(P<0.01)。结论疏肝健脾方药可使NAFLD大鼠肝组织中UCP2基因和蛋白表达水平降低。

Objective To observe effects of soothing liver and invigorating spleen recipes on expression levels of uncoupling protein 2 （UCP2） mRNA and UCP2 protein in hepatic tissues of rats with non-alcoholic fatty liver disease（NAFLD）, to preliminarily explore the possible mechanism. Methods Male SD rat model of NAFLD was induced by feeding with high fat diets for 12 weeks. Then model rats were averagely subdivided into 5 subgroups： Soothing Liver group, Invigorating Spleen group, Soothing Liver and Invigorating Spleen group （Integrated group）, Sanqi Zhigan Wan （pellets including Panax pseudo ginseng for fatty liver） group （Sanqi group）, and model group. After treated correspondingly for eight weeks, serum levels of total cholesterol, triglyceride, low density lipoprotein-cholesterol and high density lipoprotein cholesterol of each group were assayed. Expression levels of UCP2 mRNA and UCP2 protein in hepatic tissues were assayed using reverse transcription polymerase chain reaction and immunohistochemistry, respectively. Results Compared with that of normal group, the expression level of UCP2 mRNA in rat he patio tissues of model group was significantly increased （P〈0.05）. Compared with that of model group, the expression level of UCP2 mRNA in Soothing Liver group, Invigorating Spleen group and Integrated group was decreased （P〈0.05）. The expression level of UCP2 protein was significantly increased in the model group. Compared with that of model group, expression levels of UCP2 protein in treated groups were significantly decreased （P〈0. 01）. Conclusion Soothing liver and invigorating spleen recipes can down-regulate expression levels of UCP2 mRNA and UCP2 protein in hepatic tissues of NAFLD rats.