沉默Fas和PKCδ表达对游离脂肪酸诱导的HUVEC凋亡的影响  

Role of PKCδ and Fas in free fatty acid-induced HUVEC cell apoptosis

作　　者：蔡威[1] 周逸[1] 杨锴[1] 陈曼华[1] 杨飞燕[1] 

机构地区：[1]华中科技大学同济医学院附属武汉中心医院心内科,武汉430014

出　　处：《郑州大学学报（医学版）》2014年第6期811-815,共5页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的:探讨Fas与PKCδ在游离脂肪酸(FFA)诱导的人脐静脉内皮细胞(HUVEC)凋亡中的作用。方法:1取对数生长期HUVEC,分为空白组,对照组,50、75、100μmol/L FFA组及相应FFA+PKCδsiRNA转染组,采用比色法检测细胞增殖情况,Western blot检测p-PKCδ蛋白的表达。2将HUVEC分为空白组,对照组,FFA组(100μmol/L FFA处理)和FFA+PKCδsiRNA组,采用流式细胞术检测细胞凋亡情况。3取100μmol/L FFA干预后的HUVEC分别转染无关序列siRNA和Fas siRNA,以未转染细胞作空白对照,Western blot法检测p-PKCδ和Fas蛋白的表达。结果:1各组细胞增殖水平差异有统计学意义(F=20.863,P<0.001),FFA可抑制细胞增殖(P<0.05),而PKCδsiRNA能减弱FFA对细胞增殖的抑制(P<0.05)。各组细胞p-PKCδ蛋白的表达差异有统计学意义(F=229.072,P<0.001),FFA能提高p-PKCδ蛋白的表达水平(P<0.05),而PKCδsiRNA可抑制FFA引起p-PKCδ蛋白的表达(P<0.05)。2各组细胞凋亡率差异有统计学意义(F=86.973,P<0.001),FFA可增加细胞凋亡(P<0.05),而PKCδsiRNA能降低FFA所致的凋亡(P<0.05)。3各组细胞Fas蛋白表达差异有统计学意义(F=122.162,P<0.001),Fas siRNA可减低HUVECFas蛋白的表达(P<0.05)。结论:FFA诱导的HUVEC凋亡可能由PKCδ涉及的Fas通路介导。Aim：To investigate the effect of PKCδand Fas on free fatty acid-induced apoptosis in human umbilical vein endothelium cell （ HUVEC） .Methods：PKCδsiRNA was transfected into HUVEC , and RT-PCR was used to detect the ex-pression of PKCδ mRNA.HUVEC cells were treated with blank , control, 50,75,100 μmol/L FFA and the corresponding PKCδsiRNA transfected groups .FFA groups were treated with 50, 75 and 100 μmol/L FFA, and PKCδ siRNA transfected groups were transfected with PKCδsiRNA after treatment with different concentrations of FFA .Colorimetric assay was used to determine cell proliferation , and Western blot was used to investigate the protein expression of p-PKCδ.HUVEC cells were classified into four groups：blank, control, FFA and FFA＋PKCδ siRNA group.FFA group was treated with 100 μmol/L FFA, and FFA＋PKCδsiRNA group was transfected with PKCδsiRNA after treated with 100μmol/L FFA.Flow cytometry was used to measure the apoptosis rate of the four groups .100μmol/L FFA treated HUVEC cells were transfected with unre-lated sequence siRNA and Fas siRNA ,the FFA treated HUVEC cells without siRNA transfection was used as the control . Western blot was used to investigate the protein expression of p-PKCδand Fas.Results：The mRNA expression of PKCδwas decreased in cells transfected with PKCδsiRNA （F=36.839, P〈0.001）.Significant statistical difference was observed in cell proliferation between groups （F=20.863,P〈0.001）.FFA could significantly inhibit the proliferation of HUVEC cells （P〈0.05）, whereas PKCδsiRNA decreased the effect of FFA in HUVEC cells （P〈0.05）.Significant statistical difference was observed in protein expression of p-PKCδbetween groups （F=229.072, P〈0.001）.FFA could upregulate the protein expression of p-PKCδ（P〈0.05）, while PKCδsiRNA could abolish this effect （P〈0.05）.Significant statistical difference was observed in cell apoptosis between groups （F=86.973,P〈0.001）.FFA could increase cell apoptosis （P〈0.05）, while PK

关 键 词：PKCΔ Fas游离脂肪酸 人脐静脉内皮细胞 凋亡 SIRNA 

分 类 号：R363[医药卫生—病理学]