去整合素金属蛋白酶10在糖尿病冠状动脉支架内再狭窄中作用的研究  

Effects of a disintegrin and metalloprotease 10 on coronary artery in-stent restenosis in diabetes

作　　者：吴丽苹[1] 王燕萍[2] 蔡沁[2] 陈媛媛[2] 杨克[2] 陆林[2] 沈卫峰[2] 曹久妹[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院老年病科,200025 [2]上海交通大学医学院附属瑞金医院心内科,200025

出　　处：《国际心血管病杂志》2017年第2期105-109,共5页International Journal of Cardiovascular Disease

基　　金：国家自然科学基金(81570226)

摘　　要：目的:探讨去整合素金属蛋白酶10(ADAM10)在糖尿病冠状动脉(冠脉)支架内再狭窄(ISR)中的作用。方法:在17头糖尿病猪和10头正常猪的冠脉内置入雷帕霉素洗脱支架,6个月后行冠脉造影,留取发生和未发生ISR的冠脉组织,Western blot检测ADAM10表达水平。在人主动脉平滑肌细胞(HASMC)中感染ADAM10的过表达和敲减病毒,BrdU检测细胞增殖,划痕实验检测细胞迁移能力。分别用低糖培养基、高糖培养基、晚期糖基化终末产物-牛血清白蛋白(AGE-BSA)、AGE-BSA+AGE受体(RAGE)抗体培养HASMC,实时定量RT-PCR和Western blot检测ADAM10表达水平。结果:在糖尿病组中,未发生ISR和发生ISR的冠脉组织中ADAM10的表达均高于非糖尿病组(3.36±1.27对2.11±2.05,10.48±4.72对6.72±1.36,P均<0.01)。在非糖尿病组和糖尿病组,发生ISR的冠脉组织中ADAM10的表达均高于未发生ISR的冠脉组织(P均<0.01)。BrdU实验显示,在低糖培养基和高糖培养基中,ADAM10过表达的HASMC增殖均明显高于转染空载体的HASMC(2.25±0.07对1.87±0.08,2.47±0.10对2.07±0.10,P均<0.05);而ADAM10敲减的HASMC增殖均明显低于转染空载体的HASMC(1.34±0.10对1.87±0.08,1.46±0.09对2.07±0.10,P均<0.05);ADAM10过表达和ADAM10敲减的HASMC在高糖培养基中的增殖均明显高于低糖培养基(P均<0.05)。细胞划痕实验显示,在低糖培养基和高糖培养基中,ADAM10过表达的HASMC迁移距离均明显大于转染空载体的HASMC[(1.02±0.12)mm对(0.65±0.04)mm,(1.26±0.06)mm对(0.78±0.06)mm,P均<0.05)],而ADAM10敲减的HASMC迁移距离均明显小于转染空载体的HASMC[(0.26±0.06)mm对(90.65±0.04)mm,(0.43±0.14)mm对(0.78±0.06)mm,P均<0.05)];ADAM10过表达和ADAM10敲减的HASMC在高糖培养基中的迁移距离均明显大于低糖培养基(P均<0.05)。与低糖培养基相比,高糖培养基和AGE-BSA中HASMC ADAM10 mRNA和蛋白的相对表达水平均明显升高(P均<0.05);与AGE-BSA相比,AGE-BSA+RAGE抗体中HASMC ADAM10 mRNA�Objective:To investigate the effects of a disintegrin and metalloprotease 10(ADAM10)on coronary artery in-stent restenosis(ISR)in diabetes. Methods:Rapamycin-eluting stents were implanted in the coronary arteries of 17 diabetic and 10 normal minipigs,and angiography was repeated after 6 months.The coronary artery tissues of significant ISR and non-ISR segments in both diabetic and normal minipigs were analyzed by western blot analysis to detect the expression of ADAM10.Overexpression and konckdown of ADAM10 were transfected by retrovirus in human aortic smooth muscle cells(HASMC).The proliferation of HASMC was measured by BrdU assay and migration was detected by scratch test.The ADAM10 expressions of HASMC were analyzed by real time RT-PCR and western blot after treatment with low glucose,high glucose,advanced glycation end products-bovine serum albumin(AGE-BSA),and AGE-BSA+receptor for AGE(RAGE)antibody. Results:The results showed that the expressions of ADAM10 in both non-ISR tissues(3.36±1.27 vs.2.11±2.05,P<0.01)and ISR tissues(10.48±4.72 vs.6.72±1.36,P<0.01)were significantly higher in diabetic minipigs than those in normal minipigs.ADAM10 levels were significantly increased in ISR tissues compared with non-ISR tissues in both normal minipigs and diabetic minipigs(both P<0.01).BrdU assay showed that The proliferation of HASMC with overexpre-ssion of ADAM10 increased in both low glucose culture(2.25±0.07 vs.1.87±0.08,P<0.05)and high glucose culture(2.47±0.10 vs.2.07±0.10,P <0.05)compared with that of HASMC transfected by empty vector,while the proliferation of HASMC with knockdown of ADAM10 significantly inhibited in both low glucose culture(1.34±0.10 vs.1.87±0.08,P<0.05)and high glucose culture(1.46±0.09 vs.2.07±0.10,P<0.05)compared with that of HASMC transfected by empty vector.The cell proliferation in high glucose culture was significantly higher than that in low glucose culture both in HASMC with overexpression and knockdown of ADAM10(both P<0.05).Cell scratch test showed that the cell migrati

关 键 词：去整合素金属蛋白酶10 支架内再狭窄 平滑肌细胞 糖尿病 

分 类 号：R543.3[医药卫生—心血管疾病] R587.1[医药卫生—内科学]