熊果酸对肝星状细胞内AP-1、NF-κB表达的影响  

Effect of ursolic acid on AP-1 and NF-κB expression in hepatic stellate cells

作　　者：陈涛[1] 何文华[1] 黄雯[1] 余珊珊[1] 陈标[1] 黄德强[1] 李弼民[1] 朱萱[1] 

机构地区：[1]南昌大学第一附属医院消化内科,南昌330006

出　　处：《广东医学》2016年第17期2545-2548,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81160061;81260082)

摘　　要：目的明确NOX对血管紧张素II(AngⅡ)诱导的HSC转录因子激活蛋白-1(AP-1)和核因子-κB(NF-κB)的调控作用及熊果酸(UA)干预后的影响。方法将培养激活的HSC-T6细胞株分为:AngⅡ组,给予AngⅡ(1μmol/L)刺激细胞;空白对照组:不加任何药物;各干预组分别给予UA(50μmol/L)、NOX抑制剂DPI(20μmol/L)预处理30 min,再加入AngⅡ处理不同时间。采用活性氧检测试剂盒和荧光酶标仪检测其余各组细胞内荧光强度;用电泳迁移率(EMSA)测定细胞内AP-1、NF-κB的活性。结果 HSC-T6经药物作用30 min后,AngⅡ组DCF荧光强度比空白对照组明显升高(P<0.05),分别给予UA、DPI干预后细胞内DCF荧光强度均显著低于AngⅡ组(P<0.05),AngⅡ+UA组与AngⅡ+DPI组相比较DCF荧光强度无明显差异(P>0.05);用AngⅡ刺激HSC-T6细胞1 h后,AngⅡ组AP-1、NF-κB的活性明显高于空白对照组(P<0.05),分别给予UA、DPI干预后,细胞内AP-1、NF-κB的活性均显著低于AngⅡ组;AngⅡ+UA组与AngⅡ+DPI组相比较AP-1、NF-κB的活性无明显差异(P>0.05)。结论 NOX产生的ROS介导AngⅡ刺激HSC转录因子AP-1和NF-κB的活化,熊果酸通过抑制HSC内ROS的生成阻断AP-1、NF-κB的活化。Objective To clarify the effects of NOX on the generation of ROS and the activity of AP - 1 and NF - κB in rat hepatic stellate cells （ HSC - TS） induced by Ang Ⅱ. Methods Culture - activated HSC - T6 cells were divided as followed ： Ang Ⅱ group （ received Ang Ⅱ） ; normal control group （ received no treatment ） ; and intervention groups, in which HSC -T6 cells were pretreated with UA （50 p.moL/L） and NOX inhibitor DPI （20 μmol/L） for 30 min, and subsequently stimulated with Ang Ⅱ for different time periods. The fluorescence intensity was examined with ROS detection kit and fluorescence microplate. The activities of AP - 1 and NF - κB were determined using electrophoretic mobility shift assay （EMSA）. Results After treated with Ang Ⅱ for 30 rain, the fluorescence intensity was higher than control group （ P 〈 0. 05 ）. After intervened by DPI and UA, the fluorescence intensity was significantly lower than that in Ang Ⅱ treated groups （ P 〈 0.05 ）. After treated with Ang Ⅱ for one hour, the activities of AP - 1 and NF - κB were significantly higher than control group （ P 〈 0.05 ）. After intervened by DPI and UA, The activities of AP - 1 and NF - κB were significantly lower than that in Ang Ⅱ treated groups （ P 〈 0. 05 ）. Conclusion NOX can mediate the activation of AP - 1 and NF - κB by influencing the generation of ROS, UA Can reduce the activity of AP - 1 and NF - κB by inhibiting generation of ROS.

关 键 词：肝星状细胞 血管紧张素Ⅱ 熊果酸 活性氧 激活蛋白-1 核因子-ΚB 

分 类 号：R378[医药卫生—病原生物学]