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

摘要:

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、１０ μmol/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（In－tercellular 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、1 μmol/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。

Abstract:

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 activ－ity of nuclear factor-κB（NF-κB） was detected by electrophoretic mobility shift assay（EMSA）. Results：Compared with those in nor－mal control group，apoptosis rates of HUVECs were increased significantly，gene expressions of inflammatory factors ICAM-1 and IL-6 were increased，activity of NF-κB was increased significantly in AN group and A/R group（P<0.05）. 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（P<0.05），gene expressions of ICAM-1 and IL-6 were inhibited in A/R+T 0.1 group and A/R+T 1 group（P<0.05），moreover，activity of NF-κB was decreased significantly in A/R+T 0.1 group and A/R+T 1 group（P<0.05）. Compared with those in A/R+T 0.1 group，cell apoptosis rates，gene expressions of ICAM-1 and IL-6 and activity of NF-κB were reduced significantly in A/R+T 1 group（P<0.05）. 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.