摘要
目的:通过研究鳖甲煎丸药物血清对大鼠肝星状细胞(HSC-T6)中β-catenin的表达水平及NF-κB信号通路的活化的调控作用,探讨鳖甲煎丸抗肝纤维化的作用机制。方法:鳖甲煎丸(0.45 g/kg,0.9 g/kg,1.8 g/kg)和秋水仙碱(0.2 mg/kg)药物血清培养永生化HSC-T6细胞,培养24 h后采用q PCR检测HSC-T6细胞中β-catenin m RNA的表达水平,免疫荧光检测β-catenin蛋白的表达水平,Western blotting检测p65、p50的表达水平,ELISA检测下游靶基因TGF-β和TNF-α的表达水平。结果:与空白对照组相比,鳖甲煎丸(0.45g/kg、0.9g/kg、1.8g/kg)和秋水仙碱(0.2 mg/kg)药物血清组中β-catenin m RNA的表达水平显著降低,同时,鳖甲煎丸药物血清组(0.45g/kg、0.9g/kg、1.8g/kg)能够抑制p65的蛋白表达水平,但对p50的的表达无显著影响,并可显著抑制TGF-β和TNF-α的表达,且这种调控作用具有浓度依赖性。结论:鳖甲煎丸抗肝纤维化的药理作用可能与其下调HSC-T6细胞中β-catenin的表达水平,调控NF-κB信号通路的活化,并有效抑制下游靶基因TGF-β和TNF-α的表达水平密切相关,这可能是鳖甲煎丸抑制肝星状细胞增殖、活化的重要分子机制之一。
Objective： To explore the molecular mechanism of Biejiajian Pills on inhibiting hepatic fibrosis by studying its influence on the expres- sion of β-catenin and the activation of NF-κB signaling pathway. Methods： HSC-T6 cells were cultured with Biejiajian Pills （0.45g/kg,0. 9g/kg, 1. 8g/kg）and colchicine tablets （0.2mg/kg） containing serum. After 24 hours, the expressions of β-catenin mRNA were determined by qPCR ; the expression of β-catenin protein was measured by immunofluorescence ; the expressions of 1365,p50 were determined by Western Blot ;the expressions of TGF-β and TNF-α were determined by ELISA. Results： in groups of Biejiajian Pills（ 0.45g/kg,0.9g/kg, 1. 8g/kg） and colchicine tablets（0.2mg/kg）, the expressions of β-catenin mRNA and p65 protein were significantly lower than that in control group, with also inhibition of TGF-β and TNF-α ,and the effect was found in a dose-dependent manner,but showed no significant influence on the ex- pressions of p50. Conclusions： Biejiajian Pills can effectively inhibit the activation and proliferation of HSC, which is closely related to down- regulating expressions of β-catenin and regulating activation of NF-κB signaling pathway and inhibit the expressions of TGF-β and TNF-α.
出处
《中药药理与临床》
CSCD
北大核心
2017年第2期2-6,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金项目(基金号:81373807)
