【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。更多还原

【Abstract】 AIM:To investigate the effects of Shu-gan Jian-pi prescriptions on the expression of inhibtor κB 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 recipes,Jian-pi recipes and composite recipes.After intragstrically treated with the recipes for 16 weeks,the rats were executed 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 automatic biochemical analyzer.The histopathological changes of the liver tissues were observed under microscope with HE staining.The serum levels of TNF-α,IL-1 and IL-6 were determined by ELISA.The mRNA expression of IKKβ in 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 tissues 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-α,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-α,IL-1 and IL-6 in the hepatic homogenate in composite recipe high dose group and Jian-pi recipe high dose group were significantly decreased.The expression of IKKβ mRNA and p-IKKβ protein in composite recipe high and low dose groups and Jian-pi recipe high dose group were also decreased.CONCLUSION: The increasing levels of inflammatory factors and high expression of liver IKKβ mRNA/p-IKKβ protein play roles in the development of NASH.Reducing the accumulation of inflammatory factors and down-regulation of liver IKKβ may be an important mechanism of Shu-gan Jian-pi prescriptions to prevent and cure NASH.更多还原