人剪切修复基因D介导ox-LDL促进HUVEC凋亡作用  

Hxeroderma pigmentosum mediates the effect of ox-LDL on promoting the apoptosis of HUVEC

作　　者：俞菊梅 胡利琳[2] 丁颖[3] 元文峰 李菊香[5] 孙国芳[3] YU Ju-Mei;HU Li-Lin;DING Ying;YUAN Wen-Feng;LI Ju-Xiang;SUN Guo-Fang(Department of ECG, the First Affiiated Hospital of Nanehang University, Nanchang, Jiangxi 330006, China;Medical school of Nanchang University, Nanehang, Jiangxi 330006, China;Department of ECG Diagnosis, the Second Affliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, China;People＇ s Hospital of Le＇ An County, Fuzhou, Jian- gxi 344300, China;Deptartment of Cardiovascular, the Second Affliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, China)

机构地区：[1]南昌大学第一附属医院心电图室,江西省南昌市330006 [2]南昌大学医学院,江西省南昌市330006 [3]南昌大学第二附属医院心电诊断室,江西省南昌市330006 [4]江西省乐安县人民医院,江西省抚州市344300 [5]南昌大学第二附属医院心血管内科,江西省南昌市330006

出　　处：《中国动脉硬化杂志》2018年第1期19-24,共6页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81300348);江西省青年科学基金资助项目(20132BAB215015);江西省卫生厅科技计划项目(20143084)

摘　　要：目的 探讨人剪切修复基因D（XPD）是否介导氧化型低密度脂蛋白（ox-LDL）促进人脐静脉内皮细胞（HUVEC）凋亡。 方法 以ox-LDL建立HUVEC凋亡模型。用脂质体将XPD-siRNA转染HUVEC,给予ox-LDL处理。实验分6组：空白对照组；阴性对照siRNA组；XPD-siRNA组；ox-LDL组；ox-LDL ＋阴性对照siRNA组；ox-LDL＋XPD-siRNA组。MTT测定细胞活力；流式细胞仪检测细胞凋亡率和细胞周期；用RT-PCR和Western blot检测XPD、Bax和Bcl-2的表达。 结果 建立HUVEC凋亡模型ox-LDL的最佳浓度为100 mg/L。与阴性对照siRNA组相比,XPD-siRNA组的细胞凋亡率明显下降（P〈0.05）,存活率明显增加（P〈0.01）,G0/G1期细胞减少（P〈0.05）、S期细胞增加（P〈0.05）,XPD、Bax表达降低（P均〈0.05）,Bcl-2表达增高（P〈0.05）；与空白对照组相比,ox-LDL组细胞凋亡率明显增加（P〈0.01）,细胞存活率下降（P〈0.05）,G0/G1期细胞增加（P〈0.05）、S期细胞明显减少（P〈0.01）,XPD、Bax表达升高（P均〈0.05）,Bcl-2表达降低（P〈0.05）；与ox-LDL＋阴性对照siRNA组相比,ox-LDL＋XPD-siRNA组细胞凋亡率下降（P〈0.05）,细胞存活率明显增加（P〈0.01）,G0/G1期细胞减少（P〈0.05）、S期细胞增加（P〈0.05）,XPD、Bax表达降低（P均〈0.05）,Bcl-2表达增高（P〈0.05）。 结论 XPD能介导ox-LDL促进HUVEC凋亡作用。Aim To explore whether the hxeroderma pigmentosum D（XPD） could mediate the apoptosis of human umbilical vein endothelial cells （HUVEC） promoted by oxidized low density lipoprotein （ox-LDL）. Methods The model of apoptosis of HUVEC was established by ox-LDL. XPD-siRNA was transfected into HUVEC with liposome,followed by treatment with ox-LDL. This experiment was divided into six groups：blank control group; negative control siRNA group; XPD-siRNA group; ox-LDL group; ox-LDL＋negative control siRNA group; ox-LDL＋XPD-siRNA group. Cell vitality was detected by MTT; Cell apoptosis rate and cell cycle were assessed with flow cytometry; The expressions of the XPD, Bax and Bcl-2 were detected by RT-PCR and Western blot. Results The optimal concentration of ox-LDL to establish the model of apoptosis of HUVEC was 100 mg/L. Compared with negative control siRNA group, the cell apoptosis rate of XPD-siRNA group was significantly decreased （P〈0.05）, the survival rate was significantly increased （P〈0.01）, the cell number in G0/G1 phase decreased （P〈0.05）, while increased in S phase （P〈0.05）, the expressions of XPD and Bax were declined （all P〈0.05）, while the expression of Bcl-2 was elevated （P〈0.05）; Compared with blank control group, the cell apoptosis rate of ox-LDL group increased significantly （P〈0.01）, the survival rate was decreased （P〈0.05）, the cell number in G0/G1 phase increased （P〈0.05）,while decreased significantly in S phase （P〈0.01）, the expressions of XPD and Bax were elevated （all P〈0.05）,while the expression of Bcl-2 was declined （P〈0.05）. Compared with ox-LDL＋negative control siRNA group,the cell apoptosis rate of ox-LDL＋XPD-siRNA group was reduced （P〈0.05）,the survival rate was increased（P〈0.01）, the cell number in G0/G1 phase decreased （P〈0.05）,while increased in S phase （P〈0.05）, the expressions of XPD and Bax were declined （all P〈0.05）, while the expression of Bcl-2 was elevated （P〈

关 键 词：人剪切修复基因D RNA干扰 人脐静脉内皮细胞 氧化型低密度脂蛋白 细胞凋亡 

分 类 号：R34[医药卫生—基础医学]