目的探讨肝X受体(LXR)激动剂T0901317对人。肾小球内皮细胞血栓调节蛋白(TM)表达的影响及可能机制。方法不同浓度的T0901317刺激人。肾小球内皮细胞不同时间后,采用Western印迹检测LXRa、LXRβ表达;实时定量PCR、Western印迹及免疫荧光法检测T0901317对TMmRNA和蛋白表达的影响。LXRa、LXRβ基因干扰片段Si—hLXRa、Si—hLXRβ均以100nmol/L的浓度转染人肾小球内皮细胞,采用Western印迹、实时定量PCR法检测si—hLXR、Si—hLXRβ对TM蛋白和mRNA表达的影响。结果人肾小球内皮细胞表达LXRa、LXRa。与正常细胞组及DMSO对照组相比,T0901317可促进人肾小球内皮细胞TM表达(P〈0.05),且呈时间和剂量依赖性。Si—hLXRa组TM表达显著低于对照组(P〈0.05),而Si—hLXRβ组TM表达与对照组差异无统计学意义。结论人。肾小球内皮细胞表达LXRa和LXRβLXR激动剂T0901317可能主要是通过激活LXRa促进人肾小球内皮细胞TM表达。
Objective To explore the role of liver X receptor (LXR) agonist T0901317 on thrombomodulin (TM) expression in human glomerular endothelial cells and the possible mechanisms. Methods Different concentrations of T0901317 were used to stimulate human glomerular endothelial cells for different time, then LXRoL, LXRβ expression were detected by using Western blotting analysis; the roles of T0901317 on TM mRNA and TM protein expression were observed by using real-time PCR, Western blotting and immunofluorescence assay. LXRa, LXRβ gene interference segment Si-hLXRa, Si -hLXR were trausfected into human glomerular endothelial cells with the concentration of 100 nmol/L respectively, then the roles of Si-hLXRa, Si-hLXRβ on the TM protein and TM mRNA expression were assayed by Western blotting and real time PCR. Results Human glomerular endothelial cells expressed LXRa and LXR. Compared to the normal cells and DMSO group, T0901317 could significantly promote TM expression in human glomernlar endothelial cells (P 〈 0.05) and showed a time - and dose- dependent manner. TM expression in Si- hLXRa transfected group was significantly lower than that in the control group (P 〈 0.05), while TM expression in Si-hLXRβ transfected group had no significant difference compared to the control group. Conclusions Human glomerular endothelial ceils express LXRa and LXR. LXR agonist T0901317 promotes TM expression in human glomerular endothelial cells, which may be mainly through activating LXRa.