[1]蔡威,周逸,杨锴,等.沉默Fas和PKCδ表达对游离脂肪酸诱导的HUVEC凋亡的影响[J].郑州大学学报(医学版),2014,(06):811.
 CAI Wei,ZHOU Yi,YANG Kai,et al.Role of PKCδ and Fas in free fatty acidinduced HUVEC cell apoptosis[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2014,(06):811.
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沉默Fas和PKCδ表达对游离脂肪酸诱导的HUVEC凋亡的影响
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2014年06期
页码:
811
栏目:
论著
出版日期:
2014-11-20

文章信息/Info

Title:
Role of PKCδ and Fas in free fatty acidinduced HUVEC cell apoptosis
作者:
蔡威周逸杨锴陈曼华杨飞燕#
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
Author(s):
CAI WeiZHOU YiYANG KaiCHEN ManhuaYANG Feiyan
Department of Cardiology,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014
关键词:
PKCδFas游离脂肪酸人脐静脉内皮细胞凋亡siRNA
Keywords:
PKCδFasfree fatty acidhuman umbilical vein endothelium cellapoptosissiRNA
分类号:
R363
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、100 μmol/L FFA组及相应FFA+PKCδ siRNA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(100 μmol/L FFA处理)和FFA+PKCδ siRNA组,采用流式细胞术检测细胞凋亡情况。③取100 μmol/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PKCδ siRNA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PKCδ siRNA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PKCδ siRNA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。
Abstract:
Aim: To investigate the effect of PKCδ and Fas on free fatty acidinduced apoptosis in human umbilical vein endothelium cell(HUVEC). Methods: PKCδ siRNA was transfected into HUVEC, and RTPCR was used to detect the expression of PKCδ mRNA.HUVEC cells were treated with blank, control, 50,75,100 μmol/L FFA and the corresponding PKCδ siRNA transfected groups.FFA groups were treated with 50, 75 and 100 μmol/L FFA, and PKCδ siRNA transfected groups were transfected with PKCδ siRNA 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+ PKCδ siRNA group was transfected with PKCδ siRNA after treated with 100 μmol/L FFA. Flow cytometry was used to measure the apoptosis rate of the four groups. 100 μmol/L FFA treated HUVEC cells were transfected with unrelated 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PKCδ and Fas. Results: The mRNA expression of PKCδ was decreased in cells transfected with PKCδ siRNA (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 PKCδ siRNA decreased the effect of FFA in HUVEC cells (P<0.05). Significant statistical difference was observed in protein expression of pPKCδ between groups (F=229.072, P<0.001). FFA could upregulate the protein expression of pPKCδ (P<0.05), while PKCδ siRNA 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 PKCδ siRNA could abolish FFA induced apoptosis (P<0.05). Significant statistical difference was observed in protein expression of Fas between groups (F=122.162, P<0.001). Fas siRNA could significantly downregulate the protein expression of Fas in HUVEC cells (P<0.05). Conclusion: FFAinduced apoptosis in HUVEC cells may possibly mediate by PKCδ and involve in the upregulation of its downstream Fas.

参考文献/References:

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备注/Memo

备注/Memo:
#通讯作者,女,1976年4月生,博士,副主任医师,研究方向:冠心病发病机制,Email:feiyanyang@hotmail.com
更新日期/Last Update: 2014-11-26