CAJ | 학술논문

目的观察缺血预适应(ischemic preconditioning,IPC)对老年大鼠心肌Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶(a disintesrin and metalloprotease with thrombospondin type 1motifs,ADAMTS-1)表达及ADAMTS-1特异小于扰核糖核酸(small interfering ribonucleic acid,siRNA)干预对老年IPC保护作用的影响. 方法成年(4月龄)和老年(24月龄)SD大鼠各32只,按抽签法分别随机分入IPC组20只和假手术组12只,IPC后留取缺血再灌注部位心肌行免疫组织化学和免疫蛋白印迹检测ADAMTS-1表达.另老年SD大鼠110只,随机分人ADAMTS-1 siRNA组55只和对照组55只,观察ADAMTS-1 siRNA转染对IPC后ADAMTS-1蛋白表达的影响,同时观察ADAMTS-1 siRNA转染对IPC后心肌梗死存活率、心功能和心肌梗死面积的影响. 结果成年和老年大鼠IPC后24 h时缺血再灌注部位心肌ADAMTS-1蛋白表达明显升高(P＜0.05),同时老年大鼠心肌的ADAMTS-1蛋白表达高于成年大鼠(P＜0.05).成年大鼠IPC后0 h和24 h时ADAMTS-1蛋白表达的免疫组织化学检测吸光度分别为0.05±0.01和0.12±0.03,免疫蛋白印迹检测吸光度为0.68±0.16和1.17±0.21.老年大鼠IPC后0 h和24 h时ADAMTS-1蛋白表达的免疫组织化学检测吸光度分别为0.07±0.03和0.21±0.04,免疫蛋白印迹检测吸光度为0.76±0.21和1.48±0.17.ADAMTS-1 siRNA干预抑制老年大鼠IPC后的ADAMTS-1蛋白表达(IPC后0 h和24 h时的ADAMTS-1免疫蛋白印迹检测吸光度分别为0.66±0.19和0.78±0.21,P＞0.05),但不影响老年IPC大鼠心肌梗死存活率[ADAMTS-1 siRNA组和对照组老年大鼠分别为14.3%(5/35)和17.1%(6/35),P＞0.05]、心肌梗死面积[分别为(39.0±4.1)%和(38.0±5.3)%,P＞0.05]和左心室短轴缩短率[分别为(14.0±3.2)%和(13.0±2.9)%,P＞0.05]. 结论延迟IPC促使老年心肌ADAMTS-1表达增加,ADAMTS-1 siRNA转染能够抑制ADAMTS-1的表达但不能恢复老年心肌的IPC保护作用.
목적 관찰결혈예괄응(ischemic preconditioning,IPC)대노년대서심기Ⅰ형혈소판결합단백기서적해취단백양금속단백매(a disintesrin and metalloprotease with thrombospondin type 1motifs,ADAMTS-1)표체급ADAMTS-1특이소우우핵당핵산(small interfering ribonucleic acid,siRNA)간예대노년IPC보호작용적영향. 방법성년(4월령)화노년(24월령)SD대서각32지,안추첨법분별수궤분입IPC조20지화가수술조12지,IPC후류취결혈재관주부위심기행면역조직화학화면역단백인적검측ADAMTS-1표체.령노년SD대서110지,수궤분인ADAMTS-1 siRNA조55지화대조조55지,관찰ADAMTS-1 siRNA전염대IPC후ADAMTS-1단백표체적영향,동시관찰ADAMTS-1 siRNA전염대IPC후심기경사존활솔、심공능화심기경사면적적영향. 결과성년화노년대서IPC후24 h시결혈재관주부위심기ADAMTS-1단백표체명현승고(P＜0.05),동시노년대서심기적ADAMTS-1단백표체고우성년대서(P＜0.05).성년대서IPC후0 h화24 h시ADAMTS-1단백표체적면역조직화학검측흡광도분별위0.05±0.01화0.12±0.03,면역단백인적검측흡광도위0.68±0.16화1.17±0.21.노년대서IPC후0 h화24 h시ADAMTS-1단백표체적면역조직화학검측흡광도분별위0.07±0.03화0.21±0.04,면역단백인적검측흡광도위0.76±0.21화1.48±0.17.ADAMTS-1 siRNA간예억제노년대서IPC후적ADAMTS-1단백표체(IPC후0 h화24 h시적ADAMTS-1면역단백인적검측흡광도분별위0.66±0.19화0.78±0.21,P＞0.05),단불영향노년IPC대서심기경사존활솔[ADAMTS-1 siRNA조화대조조노년대서분별위14.3%(5/35)화17.1%(6/35),P＞0.05]、심기경사면적[분별위(39.0±4.1)%화(38.0±5.3)%,P＞0.05]화좌심실단축축단솔[분별위(14.0±3.2)%화(13.0±2.9)%,P＞0.05]. 결론연지IPC촉사노년심기ADAMTS-1표체증가,ADAMTS-1 siRNA전염능구억제ADAMTS-1적표체단불능회복노년심기적IPC보호작용.
Objective To investigate the effect of ischemic preconditioning (IPC) on the expression of a disintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1), and to study whether the application of small interfering (si)RNA specifically targeting ADAMTS-1 would help to recover IPC protection in the aged heart. Methods The 32 young (4 months) and 32 aged(24 months) male Sprague-Dawley (SD) rats were assigned randomly to IPC group (n=20) and sham operated group (n= 12) respectively. Myocardial samples from the ischemic-reperfused region were harvested for detecting the ADAMTS-1 expression. In addition, the 110 aged SD rats were assignedrandomly to ADAMTS-1 siRNA group and control group (n=55, each). The effects of ADAMTS-1siRNA transfcction on the expression of ADAMTS-1 protein, myocardial infarction survival rate,heart function and myocardial infarction size after IPC were observed.Results Twenty-four hours after IPC, the ADAMTS-1 protein expression increased significantly in iscbemic-reperfused region both in young and aged rats (P＜0. 05), and the protein expression was higher in aged rats than in young rats (P＜0.05). In young-IPC group, the absorbency showed ADAMTS-1 protein expression at 0 hrs and 24 hrs after IPC were 0. 05±0.01 and 0.12±0.03 by immunohistochemical staining, and were 0.68±0. 16 and 1. 17±0.21 by Western blots respectively. In aged-IPC group, the absorbency showed ADAMTS-1 protein expression at 0 hrs and 24 hrs after IPC were 0.07±0. 03 and 0.21 ±0.04 by immunohistochemical staining, and were 0. 76±0. 21 and 1. 48±0. 17 by Western blots. In the aged rats, ADAMTS-1 siRNA transfection inhibited ADAMTS-1 protein expression (0. 66±0. 19and 0.78±0.21, by Western blots at 0 hrs and 24 hrs after IPC, P＞0.05), but didn't improve myocardial infarction survival rates [ADAMTS-1 siRNA group and sham operated group: 14.3% (5/35) vs. 17.1 %(6/35), P＞0.05], left ventricular fractional shortening [(14.0±3.2)% vs. (13.0±2.9)%, P＞0.05] and myocardial infarction size[(39.0±4.1)% vs. (38.0±5.3)%, P＞0.05].Conclusions ADAMTS-1 expression induced by IPC increases significantly in aged versus in young rats. ADAMTS-1 knockdown by siRNA inhibits ADAMTS-1 protein expression but cannot recover the age-associated loss of IPC protection.