南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (06): 790-.

• • 上一篇    下一篇

NLRP3炎症小体介导血管紧张素Ⅱ诱导的人脐静脉血管内皮细胞炎症因子IL-1β的表达

杨仁强,黄玲,马晓欣,金思一,王丹,李旭   

  • 出版日期:2016-06-20 发布日期:2016-06-20

NLRP3 inflammasome mediates angiotension II-induced expression of inflammatory factor interleukin-1β in human umbilical vein endothelial cells

  • Online:2016-06-20 Published:2016-06-20

摘要: 目的研究血管紧张素Ⅱ(AngⅡ)对人脐静脉血管内皮细胞(HUVECs)NLPR3 炎症小体激活与炎症因子白介素-1β (IL-1β)表达的影响。方法体外培养HUVECs,用AngⅡ的不同浓度和刺激时间刺激HUVECs,找到最适合的AngⅡ刺激浓度 与时间。AngⅡ刺激HUVECs前,预用AngⅡ受体阻断剂氯沙坦阻断AngⅡ作用;NAD(P)H抑制剂DPI或H2O2清除剂CAT降 低胞内活性氧(ROS)含量;用caspase-1抑制剂YVAD阻断caspase-1作用;用NLRP3 siRNA沉默胞内NLRP3表达。运用蛋白 印迹法(western blot)分别检测细胞内NOX4、NLRP3、caspase-1以及IL-1β含量。结果(1)HUVECs在浓度10-9MAngⅡ刺激 12 h后,胞内NOX4、NLRP3、caspase-1以及IL-1β的蛋白表达显著增加;(2)氯沙坦、DPI、CAT、YVAD和NLRP3 siRNA都可减 轻AngⅡ诱导的上述反应。结论AngⅡ通过促进HUVECs活性氧生成,激活NLRP3炎症小体,促进胞内炎症介质IL-1β P17活 性片段生成增加,诱导血管炎症的发生。

Abstract: Objective To investigate the effect of angiotension II (AngII) on the activation of NLRP3 inflammasome and the expression of interleukin-1β (IL-1β) in human umbilical vein endothelial cells (HUVECs). Methods HUVECs cultured in vitro were treated with different concentrations of AngII for varying lengths of time to determine the optimal concentration and time for AngII exposure. To test the impact of different agents on the effect of AngII exposure, HUVECs were pretreated with AngII receptor blocker losartan, NAD(P)H inhibitor DPI and H2O2 scavenger CAT, caspase 1 inhibitor YVAD, or NLRP3 siRNA for silencing NLRP3, and the protein levels of NOX4, NLRP3, caspase-1 and IL-1β in HUVECs were analyzed by Western blotting. Results AngII treatment at the optimal concentration (10-9 mol/L) for 12 h significantly increased the protein levels of NOX4, NLRP3, caspase1 and IL-1β in HUVECs. Pretreatment with losartan, DPI, CAT, YVAD, or NLRP3 siRNA all attenuated the effects of AngII on the cells. Conclusion AngII can induce vascular inflammation by promoting the production of reactive oxygen species and activating NLRP3 inflammasome to increase the protein expression of IL-1β in HUVECs.