着色性干皮病基因B对白介素-6介导的血管平滑肌细胞增殖及凋亡的影响  

Effects of xeroderma pigmentosum B gene on proliferation and apoptosis of vascular smooth muscle cells induced by interleukin-6

作　　者：丁浩[1] 李菊香[1] 洪葵[1] 孙国芳[1] 张南[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,330006

出　　处：《中华老年医学杂志》2011年第10期849-853,共5页Chinese Journal of Geriatrics

基　　金：国家重点基础研究发展计划（973计划）资助（2008CB517305）

摘　　要：目的探讨着色性干皮病基因B（xeroderma pigmentosum B，XPB）对白介素-6（IL-6）介导入血管平滑肌细胞（VSMC）增殖及凋亡的影响。方法用脂质体转染法将重组质粒pcDNA3．1-XPB和空载质粒pcDNA3．1稳定转染VSMC，然后给予100U／ml的IL-6孵育48h。实验分为6组：（1）空白对照组；（2）peDNA3．1组；（3）pcDNA3．1-XPB组；（4）IL-6组；（5）IL-6＋pcDNA3．1组；（6）IL-6＋pcDNA3．1-XPB组。用反转录聚合酶链反应（RT—PCR）和蛋白质免疫印迹法（Westernblotting）检测XPB、Bcl-2、Bax和野生型p53（wt—p53）表达量的变化；用比色法（MTT）观察细胞增殖活力；用流式细胞仪检测细胞周期和凋亡率。结果重组质粒pcDNA3．1-XPB转染使细胞XPB表达增高（P〈0.05或P％0．01），同时Bcl-2表达降低、Bax和wt—p53表达增高（P〈0.05或P％0.01），抑制IL-6促进VSMC的Bcb2高表达、Bax和wt-p53降表达（P％0．05或P％0．01）；XPB高表达抑制了细胞增殖活力（q=2．95，P〈0．05），并抑制IL-6促进VSMC增殖，IL-6＋pcDNA3．1-XPB组和IL-6＋pcDNA3．1组VSMC存活率分别为（102．6±6．2）％和（124．5±7．9）％（q=3．49，P〈0．05）；流式细胞仪检测结果显示，XPB高表达引起细胞G0／G1期增加、S期减少、凋亡率增加（q值分别为2．99、5．39、3．05，P〈0.05或P〈0．01），并抑制IL-6促进VSMC G0／G1期减少、S期增加、凋亡率降低的作用，IL-6＋peDNA3．1-XPB组和IL-6＋peDNA3．1组分别为（70．9±6．7）％与（54．8±2．9）％、（20．2±3．6）％与（36．4±7．2）％、（5．9±2．1）％与（0.3±0．1）％（q值分别为6．91、8．54、7．53，均P〈0．01）。结论XPB基因能抑制VSMC增殖并促进其凋亡，并能抑制IL-6促进VSMC增殖和降低其凋亡的作用，有望成为治疗动脉粥样硬化的靶点。Objective To investigate effects of xeroderma pigmentosum B（XPB） gene on IL-6 induced proliferation and apoptosis in human vascular smooth muscle cells （VSMC）. Methods Recombinant plasmid pcDNA3.1-XPB and vacant vector plasmid pcDNA3.1 were transfected stably into VSMC by liposome, and these ceils were incubated with IL-6 at a 100 U/ml for 48 hours. The experiments were divided into six groups： blank control group; pcDNA3.1 group; pcDNA3.1-XPB group;IL-6 group; IL-6 ＋ pcDNA3.1 group; IL-6 ＋ pcDNA3.1-XPB group. The expression levels of XPB, Bcl-2, Bax and wild type p53 （wt-p53） were detected through reverse transcription polymerase chain reaction （RT-PCR） and Western blotting. The cell survival, cell cycle and apoptosis were examined with 3-（ 4, 5-Dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） and flow cytometry,respectively. Results The transfection of pcDNA3.1-XPB increased the expression of XPB, Bax and wt-p53 （P〈0.05 or P^0.01）, decreased the expression of Bcl-2 （P〈0.05 or P〈 0.01）, and reduced the IL-6 induced effects on decreasing the expression of Bax and wt-p53 and increasing the expression of Bcl-2（P〈0.05 or P〈0.01）. The over expression of XPB inhibited the cell growth（q=2.95,P〈0.05）, and reduced the positive effects of IL6 on VSMC growth（102.6± 6.2）% vs. （124. 5±7.9）%,q=3.49, P〈0.05. The over expression of XPB increased the apoptosis rate of VSMC（P〈0. 01） and the cell amounts o{ G0/G1 phase（q=2.99, P〈0.05）, decreased the cell amounts of S phase（q=3.05, P〈0.05）, and reduced the IL-6 induced effects on decreasing the apoptosis rate of VSMC（5. 922.1）%vs. （0. 320.1）%,q=7.53, P〈0.01; the cell amounts of G0/ G1 phase（70. 926.7）% vs. （54. 822.9）% ,q=6.91, P〈0.01 ;and on increasing the cell amounts of Sphase（20.223.6）% vs. （36.4±7.2）%,q=8.54, P〈0.01. Conclusions XPB gene could inhibit VSMC proliferation, promote VSMC apoptosis, and reduce the effects that IL-6 promotes VSMC prolife

关 键 词：干皮病 着色性 肌 平滑 血管 增殖 细胞凋亡 

分 类 号：R543.5[医药卫生—心血管疾病]