中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2014年
4期
299-303
,共5页
丁涵露%王楠%阮一哲%汪伟%张萍%王莉
丁涵露%王楠%阮一哲%汪偉%張萍%王莉
정함로%왕남%원일철%왕위%장평%왕리
肾小球%内皮细胞%血栓调节蛋白%肝X受体
腎小毬%內皮細胞%血栓調節蛋白%肝X受體
신소구%내피세포%혈전조절단백%간X수체
Kidney glomerulus%Endothelial cells%Thrombomodulin%Liver X receptor
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LXRα、LXRβ表达;实时定量PCR、Western印迹及免疫荧光法检测T0901317对TM mRNA和蛋白表达的影响.LXRα、LXRβ基因干扰片段Si-hLXRα、Si-hLXRβ均以100 nmol/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-hLXRα、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LXRα、LXRβ.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-hLXRα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LXRα和LXRβ.LXR 激动剂T0901317可能主要是通过激活LXRα促进人肾小球内皮细胞TM表达.
目的 探討肝X受體(LXR)激動劑T0901317對人腎小毬內皮細胞血栓調節蛋白(TM)錶達的影響及可能機製.方法 不同濃度的T0901317刺激人腎小毬內皮細胞不同時間後,採用Western印跡檢測LXRα、LXRβ錶達;實時定量PCR、Western印跡及免疫熒光法檢測T0901317對TM mRNA和蛋白錶達的影響.LXRα、LXRβ基因榦擾片段Si-hLXRα、Si-hLXRβ均以100 nmol/L的濃度轉染人腎小毬內皮細胞,採用Western印跡、實時定量PCR法檢測Si-hLXRα、Si-hLXRβ對TM蛋白和mRNA錶達的影響.結果 人腎小毬內皮細胞錶達LXRα、LXRβ.與正常細胞組及DMSO對照組相比,T0901317可促進人腎小毬內皮細胞TM錶達(P<0.05),且呈時間和劑量依賴性. Si-hLXRα組TM錶達顯著低于對照組(P<0.05),而Si-hLXRβ組TM錶達與對照組差異無統計學意義.結論 人腎小毬內皮細胞錶達LXRα和LXRβ.LXR 激動劑T0901317可能主要是通過激活LXRα促進人腎小毬內皮細胞TM錶達.
목적 탐토간X수체(LXR)격동제T0901317대인신소구내피세포혈전조절단백(TM)표체적영향급가능궤제.방법 불동농도적T0901317자격인신소구내피세포불동시간후,채용Western인적검측LXRα、LXRβ표체;실시정량PCR、Western인적급면역형광법검측T0901317대TM mRNA화단백표체적영향.LXRα、LXRβ기인간우편단Si-hLXRα、Si-hLXRβ균이100 nmol/L적농도전염인신소구내피세포,채용Western인적、실시정량PCR법검측Si-hLXRα、Si-hLXRβ대TM단백화mRNA표체적영향.결과 인신소구내피세포표체LXRα、LXRβ.여정상세포조급DMSO대조조상비,T0901317가촉진인신소구내피세포TM표체(P<0.05),차정시간화제량의뢰성. Si-hLXRα조TM표체현저저우대조조(P<0.05),이Si-hLXRβ조TM표체여대조조차이무통계학의의.결론 인신소구내피세포표체LXRα화LXRβ.LXR 격동제T0901317가능주요시통과격활LXRα촉진인신소구내피세포TM표체.
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 LXRα,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.LXRα,LXRβ gene interference segment Si-hLXRα,Si -hLXRβ were transfected into human glomerular endothelial cells with the concentration of 100 nmol/L respectively,then the roles of Si-hLXRα,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 LXRα and LXRβ.Compared to the normal cells and DMSO group,T0901317 could significantly promote TM expression in human glomerular endothelial cells (P < 0.05) and showed a time -and dose-dependent manner.TM expression in Si-hLXRα 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 cells express LXRα and LXRβ.LXR agonist T0901317 promotes TM expression in human glomerular endothelial cells,which may be mainly through activating LXRa.