疏肝健脾方对NASH大鼠肝组织IKKβmRNA和蛋白表达的影响  

Effects of Shu-gan Jian-pi prescriptions on expression of IKKβ at mRNA and protein levels in liver tissues of rats with non-alcoholic steatohepatitis

作　　者：魏波[1] 杨钦河[1] 王文晶[1] 胡巢凤[2] 方梅霞[3] 刑会杰 张玉佩[1] 程少冰[1] 王彦平[2] 冯高飞[1] 何秀敏[1] 徐拥建[1] 闫海震[1] 黄进[1] 

机构地区：[1]暨南大学医学院中医系 [2]暨南大学医学院病理生理学教研室,国家中医药管理局病理生理三级实验室 [3]暨南大学医学院实验动物管理中心,广东广州510632

出　　处：《中国病理生理杂志》2012年第8期1448-1454,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30973694)

摘　　要：目的:探讨疏肝健脾方对非酒精性脂肪性肝炎(NASH)大鼠肝组织核因子κB(NF-κB)抑制蛋白激酶β(IKKβ)mRNA和蛋白表达的影响。方法:采用高脂饲料喂养建立NASH大鼠实验模型,在施以造模因素的同时各药物干预组大鼠分别灌服疏肝方(柴胡疏肝散)、健脾方(参苓白术散)和综合方(柴胡疏肝散和参苓白术散的合方)的高、低剂量进行干预,16周后各组大鼠腹主动脉采血,全自动生化分析仪检测血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)及肝组织TC、TG的含量;ELISA检测血清肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)及白细胞介素1(IL-1)的水平;HE染色观察肝组织病理变化;实时荧光定量RT-PCR检测肝组织IKKβmRNA的表达水平;Western blotting检测肝组织IKKβ蛋白磷酸化水平的变化。结果:肝组织病理染色提示大鼠NASH造模成功,与正常组比较,模型组大鼠血清TC、LDL-C、肝组织TC、TG及炎症细胞因子TNF-α、IL-1、IL-6水平均显著升高(P<0.01,P<0.05),肝组织IKKβmRNA及其磷酸化蛋白的表达水平均明显升高(P<0.01)。与模型组相比,肝组织TC、TG含量及血清TNF-α的含量以综合方高、低剂量组、健脾方高剂量组及疏肝方高剂量组下降显著(P<0.01,P<0.05);IL-1和IL-6含量均以综合方高剂量组降低显著(P<0.05),IKKβmRNA的表达水平以综合方高组及健脾方高组下调明显(P<0.05),磷酸化IKKβ蛋白的表达水平以健脾方高剂量组及综合方高、低剂量组下降最为显著(P<0.01,P<0.05)。结论:血清炎症细胞因子TNF-α、IL-1和IL-6水平的显著升高、肝组织IKKβmRNA及其磷酸化蛋白的高表达可能参与NASH的发病。降低相关炎症细胞因子的含量、抑制IKKβmRNA及蛋白磷酸化可能是疏肝健脾方抗NASH的重要机制之一。To investigate the effects of Shu - gan Jian - pi prescriptions on the expression of inhibtor KB kinase β （IKKβ） at mRNA and protein levels in the liver tissues of rats with non - alcoholic steatohepatitis （NASH）. METHODS ： The experimental model of NASH was established by feeding a high - fat diet in rats. The Shu - gan Jian - pi prescriptions of traditional Chinese medicines were prepared and divided into high dose and low dose of Shu - gan reci- pes, Jian- pi recipes and composite recipes. After intragstrically treated with the recipes for 16 weeks, the rats were exe-cuted to obtain blood samples from the ventral aorta. The serum levels of total cholesterol （TC）, triglyceride （TG）, high - density lipoprotein cholesterol （ HDL - C） and low - density lipoprotein cholesterol （ LDL - C） were detected by an au- tomatic biochemical analyzer. The histopathological changes of the liver tissues were observed under microscope with HE staining. The serum levels of TNF - a, IL - 1 and IL - 6 were determined by ELISA. The mRNA expression of IKKβ n liver tissues was analyzed by real - time fluorescence quantitative RT - PCR. The protein level of phospho - IKKβ （ p - IKKβ） in the liver tissues was examined by Western blotting. RESULTS： The histopathological changes of the liver tis- sues in the rats fed with high - fat diet for 16 weeks indicated the successful establishment of NASH model. Compared with normal group, the serum TC and LDL - C, liver TC and TG levels, and the serum levels of TNF - a, IL - 1 and IL - 6 in model group were significantly increased and the expression of IKKβ mRNA and phospho - IKKβ protein in model group were higher. Compared with model group, the levels of serum TG, and liver TC/TG in Shu - gan recipe high dose group, Jian -pi recipe high dose group and composite recipe high and low dose groups were significantly decreased. The levels of TNF - or, IL - 1 and IL - 6 in the hepatic homogenate in composite recipe high dose group and Jian - pi recipe high dose

关 键 词：非酒精性脂肪性肝炎 疏肝健脾方 IΚB激酶 大鼠 

分 类 号：R575.5[医药卫生—消化系统]