肝X受体激动剂T0901317对缺氧复氧损伤人脐静脉内皮细胞的保护及机制研究

Effects and mechanisms of LXRs agonist T0901317 on human umbilical vein endothelial cells injury induced by anoxia/reoxygenation

目的:探讨肝X受体激动剂T0901317激活肝X受体对人脐静脉内皮细胞(Human umbilical vein endothelial cells,HUVECs)缺氧复氧(Anoxia/reoxygenation,A/R)损伤的影响及其可能机制。方法:传代培养HUVECs,建立A/R模型,实验分为6组:正常对照组,单纯缺氧(Anoxia,AN)组,缺氧复氧(A/R)组,A/R+不同浓度(0.1、1、10μmol/L)肝X受体激动剂T0901317干预组(A/R+T 0.1组、A/R+T 1组、A/R+T 10组)。流式细胞术检测各组细胞凋亡率;RT-PCR方法检测各组细胞间黏附分子-1(Intercellular adhesion molecule-1,ICAM-1)、白细胞介素-6(Interleukin-6,IL-6)mRNA表达;凝胶电泳迁移率分析法检测各组核转录因子-κB(Nuclear factor-κB,NF-κB)的活性。结果:与正常对照组比较,AN组、A/R组的细胞凋亡率升高,ICAM-1、IL-6 mRNA的表达增高,NF-κB活性增强(P<0.05)。与A/R组比较,A/R+T 0.1,A/R+T 1组显著抑制A/R损伤引起的细胞凋亡,减少炎症因子ICAM-1、IL-6 mRNA的表达,并明显抑制A/R损伤造成的NF-κB活化(P<0.05)。与A/R+T 0.1组比较,A/R+T 1组细胞凋亡率,ICAM-1、IL-6的表达及NF-κB活性均降低(P<0.05)。结论:适当剂量(0.1、1μmol/L)T0901317可激活HUVECs肝X受体,对A/R引起的HUVECs损伤起保护作用,其机制可能与抑制NF-κB转录活性从而下调NF-κB通路下游炎症因子如ICAM-1,IL-6的表达有关。

Objective：To investigate the effects and mechanisms of liver X receptors（LXRs）agonist T0901317 on human umbilical vein endothelial cells（HUVECs）injury induced by anoxia/reoxygenation（A/R）.Methods：HUVECs were cultivated and divided into 6 groups randomly：normal control group,anoxia（AN）group,A/R group,A/R groups with different concentrations of T0901317（0.1,1,10 μmol/L;A/R＋T 0.1 group,A/R＋T 1 group,A/R＋T 10 group）.The apoptosis rate of HUVECs was detected by flow cytometry.The gene expressions of intercellular adhesion molecule-1（ICAM-1）and interleukin-6（IL-6）were assessed by RT-PCR.The activity of nuclear factor-κB（NF-κB）was detected by electrophoretic mobility shift assay（EMSA）.Results：Compared with those in normal control group,apoptosis rates of HUVECs were increased significantly,gene expressions of inflammatory factors ICAM-1 and IL6 were increased,activity of NF-κB was increased significantly in AN group and A/R group（P0.05）.Compared with those in A/R group,cell apoptosis rates were decreased in A/R＋T 0.1 group and A/R＋T 1 group（P0.05）,gene expressions of ICAM-1 and IL-6 were inhibited in A/R＋T 0.1 group and A/R＋T 1 group（P0.05）,moreover,activity of NF-κB was decreased significantly in A/R＋ T 0.1 group and A/R＋T 1 group（P0.05）.Compared with those in A/R＋T 0.1 group,cell apoptosis rates,gene expressions of ICAM1 and IL-6 and activity of NF-κB were reduced significantly in A/R＋T 1 group（P0.05）.Conclusions：Proper dose of T0901317（0.1,1 μmol/L）can activate LXRs and protect HUVECs from A/R injury.The mechanisms of T0901317 may associate with the inhibition of NF-κB activation and downregulation of ICAM-1 and IL-6 gene expression.