保肝宁对肝星状细胞中核转录因子-κB活性影响的实验研究

Experimental Study on Effect of Baoganning on Activity of Nuclear Transcription Factor-κB in Hepatic Stellate Cells

目的观察中药复方保肝宁对肝星状细胞(hepaticstellatecell,HSC)中核转录因子κB(nuclearfactorκB,NFκB)活性的影响及其相关机制。方法给正常Wistar大鼠灌胃保肝宁煎剂7天,制备含药血清,用MTT法检测其对肝星状细胞HSCT6生长的影响,用蛋白印迹实验(Westernblottinganalysis)检测药物作用不同时间NFκB蛋白抑制因子IκB磷酸化水平,用凝胶阻滞实验(gelshiftassay)检测药物作用30min后NFκB与DNA的结合水平。结果中药复方保肝宁对HSCT6细胞生长有显著的抑制作用;可快速抑制IκB磷酸化,30min达到最高水平,6h恢复至基础水平;可改变NFκB与DNA结合水平,使其与DNA结合能力下降。结论中药复方保肝宁能抑制IκB磷酸化,降低NFκB活性,并改变NFκB与DNA的结合水平。

Objective To investigate the effect of Baoganning （BGN） on activity of nuclear transcription factor-κB （NF-κB） in hepatic stellate cells （HSCs） and its relevant mechanisms. Methods Normal Wlstar rats were medicated with BGN decoction by garage for 7 days to prepare BGN drug-serum. The effect of BGN drugserum on HSC-T6 growth was measured by MTT assay; phosphorylation level of NF-κB inhibiting factor IκB at different time after BGN stimulation was detected by Western blotting analysis; and the binding level of NF-κB with DNA was measured 30 min after drug-serum stimulation with gel shift assay, Results BGN could significantly inhibit the HSC-T6 growth and quickly supress the phosphorylation of IκB, with the effect reached its peak at 30 min and restored to baseline level 6 h after stimulation, and reduce the binding capacity of NF-κB with DNA, Conclusion BGN can inhibit phosphorylation of IκB, restrain the activity of NF-κB and change the binding level of NF-κB with DNA.