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-T6) induced by AngⅡ.Methods Culture-activated HSC-T6 cells were divided as followed: AngⅡ group ( received AngⅡ); normal control group ( received no treatment ); and intervention groups, in which HSC-T6 cells were pretreated with UA (50 μmol/L) and NOX inhibitor DPI (20 μmol/L) for 30 min, and subsequently stimulated with AngⅡ 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 AngⅡ for 30 min, 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 AngⅡtreated groups (P<0.05).After treated with AngⅡfor one hour, the activities of AP-1 and NF-κB were sig-nificantly 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 AngⅡtreated 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 inhibi-ting generation of ROS .
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