ROS/ALOX5/NLRP3信号通路激活在低氧所致肺动脉内皮增殖中的作用  

The effect of ROS/ALOX5/NLRP3 signaling pathway on hypoxia-induced pulmonary artery endothelial cell proliferation

作　　者：胡志 马慧 范粉玲 刘文娟[1] 谢航 

机构地区：[1]西安交通大学第一附属医院结构性心脏病科,陕西西安710061

出　　处：《实用医院临床杂志》2017年第6期44-48,共5页Practical Journal of Clinical Medicine

基　　金：陕西省自然科学基础研究基金面上项目(编号:2014JM2-8171)

摘　　要：目的考察ROS/ALOX5/NLRP3信号通路相关蛋白激活在低氧所致肺动脉内皮细胞增殖中的作用。方法将人肺动脉内皮细胞分别在常氧状态(21%O_2)和低氧状态(1%O_2)下培养24、48、72及96小时,同时低氧状态下分别设立活性氧(ROS)清除剂Trolox组和ALOX5抑制剂齐留通组。采用CCK-8法检测细胞增殖率的改变,活性氧探针H_2-DCFA检测低氧状态下肺动脉内皮细胞内ROS的变化,Western blot法分别检测低氧状态及加入Trolox或齐留通干预后ALOX5、NLRP3和Caspase-1蛋白表达的变化。结果肺动脉内皮细胞在低氧作用48、72及96 h后可见增殖显著,分别达(135.6±29.4)%、(185.3±21.7)%及(212.3±39.0)%。同时,细胞内ROS在24、48及72 h可检测到显著增加,96小时后基本恢复正常。ALOX5、NLRP3和Caspase-1蛋白随低氧时间增加而表达增多。预先加入ROS清除剂Trolox和ALOX5抑制剂齐留通均可抑制NLRP3炎性小体相关蛋白NLRP3和Caspase-1的表达,且细胞增殖率也显著降低。结论 ROS/ALOX5/NLRP3信号通路在低氧所致的肺动脉内皮细胞增殖中显著激活,抑制这一通路可显著减少低氧所致的内皮细胞增殖。Objective To investigate the role of activation of ROS/ALOX5/NLRP3 signaling pathway in hypoxia-induced proliferation of pulmonary artery endothelial cells. Methods The human pulmonary arterial endothelial cells （HPAEC） were cultured in normoxia （21% O2 ） and hypoxia （ 1% O2 ） for 24,48,72 and 96 hours,respectively. At the same time,subsets of reactive oxygen species （ROS） scavenger,Trolox and ALOX5 inhibitor,Zileuton were set up. The CCK-8 method was used to detect the cell proliferation rate. The change of ROS level in HPAEC under hypoxia was detected by probe H2-DCFA. Western blot was used to detect the expres- sion of ALOX5, NLRP3 and caspase-1 protein after intervention of hypoxia, Trolox and Zileuton, respectively. Results HPAEC were significantly proliferated at 48,72 and 96 hours after hypoxia, reaching （ 135.6±29.4 ） %, （ 185. 3 ±21.7 ） % and （ 212. 3 ±39. 0） %, respectively. At the same time, intracellular RQS at 24-,48-and 72-hour was increased significantly and return to normal at 96 hour. The protein expressions of ALOX5, NLRtr3 and caspase-1 were increased with the prolonged hypoxia time. Pretreatment with Trolox and Zileuton inhibited the expressions of NLRP3 and caspase-1 in the inflammasome, and the cell proliferation rate was decreased significantly. Conclusion ROS/ALOX5/NLRP3 signaling pathway is significantly activated in hypoxia-induced proliferation of HPAEC, and inhibition of this pathway can significantly reduce hypoxia-induced endothelial cell proliferation.

关 键 词：肺动脉高压 低氧 人肺动脉内皮细胞 NLRP3炎性小体 信号通路 

分 类 号：R563[医药卫生—呼吸系统]