摘要
目的:探讨疏肝健脾方药对非酒精性脂肪性肝炎(NASH)大鼠肝细胞NF-κB信号通路相关基因及蛋白表达的影响。方法:将SD雄性大鼠随机分为8组:正常对照组、模型组、疏肝高/低剂量组、健脾高/低剂量组、合方高/低剂量组,每组15只。采用高脂饲料喂养复制大鼠NASH模型,16后检测肝脂、血脂,油红O和HE染色观察肝组织病理学变化。同时每组取6只采用离体循环灌注Ⅳ型胶原酶法分离肝细胞,Real time Q-PCR法Western blot法分别检测各组大鼠肝细胞IKKβ、NF-κB mRNA和蛋白及磷酸化IKKβ蛋白表达。结果:与正常对照组比较,模型组大鼠肝细胞存在显著的脂质代谢紊乱;血清TC,肝脂及肝细胞IKKβ、NF-κB mRNA和蛋白表达及磷酸化IKKβ蛋白表达较正常对照组均显著升高(P<0.01);与模型组比较,健脾高剂量组、合方高剂量组大鼠肝细胞IKKβ、NF-κB mRNA表达水平显著降低(P<0.05或P<0.01),各药物干预组大鼠肝细胞IKKβ、NF-κB及磷酸化IKKβ蛋白表达均显著下调(P<0.05或P<0.01),其中以健脾高剂量组、合方高剂量组下调趋势最为显著。结论:抑制肝细胞IKKβ/NF-κB信号通路相关基因及蛋白表达可能是疏肝健脾方药防治NASH的药理作用机制之一;上述蛋白可能是其药物作用的有效靶点。疏肝健脾方药对NASH的干预可能存在一定的量效关系。
Objective：To observe the effects of soothing liver and invigorating spleen recipes on NF-κB signal pathway related genes and proteins in primary hepatocytes of rats with NASH. Methods ： SD male rats were randomly divided into 8 groups ： normal, model,high/low-dose soothing liver group ,high/low-dose invigorating spleen group ,high/low-dose integrated group. 15 rats in each group. The NASH model rats were induced by feeding high-fat diet（HFD）. The treatment lasted for 16 weeks. Then TC ,TG in the liver tissue and serum were determined with automatic biochemical analyzer. HE staining and oil red O staining were operated to observe the patho- logical changes. Another 6 rats of each group were taken respectively and collagenase （TypeIV）was perfused to digest liver tissue with the circulation in vitro to separate hepatocytes. The expression levels of IKKβ, NF-κB mRNA, proteins and phosphorylated IKKβ protein in hepatocytes of rats from each group were detected by Real-time Q-PCR and Western Blotting, respectively. Results：Compared with normal group,liver histopathology was changed and levels of TC and TG were elevated in model group indicating hepatocytes had lipid accumulation and lipid metabolic disturbance obviously;The levels of serum TC, and hepatic homogenate TC, TG as well as the expression of IKKβ, NF-κB mRNA, proteins and phosphorylated IKKβ protein in hepatocytes were dramatically increased in model group （P 〈 0. 01 ）. Compared with the model group, the levels of IKKβ, NF-κB mRNA expression were decreased most significanly in the invigorating spleen （with high dose）group and the integrated group （with high dose ） （P 〈 0.01 or P 〈 0.05 ）. The expression levels of the IKKβ, NF-κB proteins and the phosphorylated IKKβ protein in hepatocytes were decreased significaniy in the treatment groups （P 〈 0. 01 or P 〈 0.05 ）, especially for the invigorating spleen （ with high dose） group and the integrated （ with high dose ）. Conclusion ： Soothing liver and invigorating spleen recipes have effect on NASH rats induced by HFD and its mechanism may be related to the suppression of IKKβ/NF-κB signal pathway related genes and proteins. And the effect probably has a dose response relationship.
出处
《中药材》
CAS
CSCD
北大核心
2013年第9期1469-1476,共8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)
关键词
非酒精性脂肪性肝炎
疏肝健脾方药
核转录因子-ΚB
原代肝细胞
Non-alcoholic steatohepatitis
Soothing liver and invigorating spleen recipes
Nuclear factor NF-B
Primary hepatocyes
