剪切修复基因XPD抑制Ox-LDL诱导人脐动脉平滑肌细胞的增殖  

Effect of xeroderma pigmentosum group D gene on proliferation of human umbilical arterial smooth muscle cells induced by Ox-LDL

作　　者：夏子荣[1] 李青[1] 夏珍[1] 李菊香[1] 洪葵[1] 吴延庆[1] 吴清华[1] 程晓曙[1] 

机构地区：[1]南昌大学第二附属医院心内科,江西南昌330006

出　　处：《中国病理生理杂志》2017年第12期2238-2244,共7页Chinese Journal of Pathophysiology

基　　金：江西省自然科学基金资助项目(No.2010GZY0254)

摘　　要：目的:探讨剪切修复基因——着色性干皮病D组基因(XPD)在氧化低密度脂蛋白(Ox-LDL)促血管平滑肌细胞增殖中的作用及机制。方法:将重组质粒pEGFP-N2/XPD利用脂质体转染人脐动脉平滑肌细胞(HUASMCs),实验分为空白对照组、空载质粒pEGFP-N2组、重组质粒pEGFP-N2/XPD组、Ox-LDL组、Ox-LDL+pEGFP-N2组和Ox-LDL+pEGFP-N2/XPD组。用MTT法和Ed U法测定各组细胞的增殖率;流式细胞术检测各组细胞周期分布;利用Western blot法检测XPD、caspase-3、Bcl-2和Bax的蛋白水平。结果:Western blot实验结果发现,与空白对照组相比,pEGFP-N2/XPD组的XPD表达增加(P<0.05),表明转染成功;MTT和Ed U检测结果显示,pEGFP-N2/XPD组的细胞增殖率较空白对照组降低(P<0.05);与Ox-LDL组比较,Ox-LDL+pEGFP-N2/XPD组细胞增殖明显被抑制(P<0.05)。流式细胞术的检测结果显示,与空白对照组比较,pEGFP-N2/XPD组的S期细胞比例明显减少(P<0.05),G0/G1期细胞比例明显增多(P<0.05);与Ox-LDL组比较,Ox-LDL+pEGFP-N2/XPD组的S期细胞比例减少(P<0.05),G0/G1期细胞比例明显增多(P<0.05)。Western blot结果显示,与对照组比较,pEGFP-N2/XPD组的cleaved caspase-3和Bax蛋白水平增加(P<0.05),Bcl-2蛋白表达降低(P<0.05);与Ox-LDL组比较,Ox-LDL+pEGFP-N2/XPD组的cleaved caspase-3和Bax蛋白水平增加(P<0.01),Bcl-2蛋白表达降低(P<0.05)。结论:XPD能抑制HUASMCs的增殖并促其凋亡,还能抑制Ox-LDL的促HUASMCs增殖作用,有可能成为抗动脉粥样硬化治疗的靶点。AIM ： To investigate the effects of xeroderma pigmentosum group D （XPD） gene on the prolifera-tion of human umbilical arterial smooth muscle cells （HUASMCs） induced by oxidized low-density lipoprotein （Ox-LDL）. METHODS ： The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome. The cells were di-vided into blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL ＋ pEGFP-N2 group and Ox-LDL ＋ pEGFP-N2/XPD group. The proliferation rate of the cells was detected by MTT and EdU assays. The apop- totic rate and cell cycle distribution were analyzed by flow cytometry. The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS： Compared with blank control group, the expression of XPD was increased in pEGFP-N2/XPD group （P 〈0.05）. According to the results of MTT and EdU assays, the cell proliferation in pEGFP-N2/ XPD group was reduced compared with blank control group （P 〈 0 .0 5 ） . Compared with Ox-LDL group, the cell prolifera-tion in Ox-LDL ＋ pEGFP-N2/XPD group was significantly inhibited （P 〈 0 .0 5 ） . According to the results of flow cytome-try, the cell proportion of S phase decreased and the G0/ O 2-phase cell proportion increased significantly in pEGFP-N2/ XPD group and Ox-LDL ＋ pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively （P 〈 0.05）. Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL ＋ pEGFP-N2/XPD group, respectively （P 〈0. 05）. CONCLUSION： XPD inhibits the proliferation of HUASMCs and promotes their apoptosis, and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs. XPD may be the target for treatment of atherosclerosis.

关 键 词：着色性干皮病D组基因 氧化低密度脂蛋白 人脐动脉平滑肌细胞 动脉粥样硬化 

分 类 号：R0[医药卫生] R363.2