CAJ | 학술논문

目的观察重组腺相关病毒介导大鼠血红素加氧酶-1基因转染对大鼠离体心肌缺血再灌注心肌细胞凋亡的影响.方法雄性SD大鼠30只随机分成3组,对照组(C组,n=6),缺血再灌注组(I/R组,n=12),腺相关病毒介导血红索加氧酶-1基因组(A-r组,n=12).基因转染3个月后,建立大鼠离体心脏Langendorff灌流模型,C组持续灌注100 min,其他各组均平衡15 rain,停灌40min与再灌注45 min,记录冠脉流量(CF),测定冠脉流出液肌酸激酶(CK)活性,测定再灌注后45 min时的心肌心肌组织超氧化物岐化酶(SOD)活性活性及丙二醛(MDA)含量,同时取每组大鼠心肌检测梗死面积、心肌细胞凋亡率以及心肌组织bax、bcl-2蛋白表达量.结果离体心肌Langendorff灌注后,与I/R组比较,A-r组在复灌后冠脉流出液CK活性降低(P＜0.01),复灌后45min后心肌MDA含量降低(P＜0.01),SOD活性增高(P＜0.01),梗死面积较小(P＜0.01),心肌组织bax表达量、心肌细胞凋亡率均显著下降,心肌组织bel-2表达量明显增加(P＜0.01).结论重组腺相关病毒血红素加氧酶1基因转染心肌后,可抑制离体缺血再灌注心肌细胞凋亡和增强心肌抗氧化能力,对大鼠离体心肌缺血再灌注损伤有显著保护作用.
목적 관찰중조선상관병독개도대서혈홍소가양매-1기인전염대대서리체심기결혈재관주심기세포조망적영향.방법 웅성SD대서30지수궤분성3조,대조조(C조,n=6),결혈재관주조(I/R조,n=12),선상관병독개도혈홍색가양매-1기인조(A-r조,n=12).기인전염3개월후,건립대서리체심장Langendorff관류모형,C조지속관주100 min,기타각조균평형15 rain,정관40min여재관주45 min,기록관맥류량(CF),측정관맥류출액기산격매(CK)활성,측정재관주후45 min시적심기심기조직초양화물기화매(SOD)활성활성급병이철(MDA)함량,동시취매조대서심기검측경사면적、심기세포조망솔이급심기조직bax、bcl-2단백표체량.결과 리체심기Langendorff관주후,여I/R조비교,A-r조재복관후관맥류출액CK활성강저(P＜0.01),복관후45min후심기MDA함량강저(P＜0.01),SOD활성증고(P＜0.01),경사면적교소(P＜0.01),심기조직bax표체량、심기세포조망솔균현저하강,심기조직bel-2표체량명현증가(P＜0.01).결론 중조선상관병독혈홍소가양매1기인전염심기후,가억제리체결혈재관주심기세포조망화증강심기항양화능력,대대서리체심기결혈재관주손상유현저보호작용.
Objective To study the effects of rat heme oxygenase-1 ( rHO-1 ) gene carried by recombinant adeno-associated virus (AAV) on isolated rat myocardial ischemia/repeffusion injury. Methods Thirty male SD rats were randomly divided into 3 groups: control ( group c, n = 6), ischemia-reperfusion group ( I/R group, n = 12) and AAV-rHO-1 group ( A-r group, n = 12). After gene delivery for 3 months, the rats were killed and hearts were isolated and perfused with Langendorff apparatus. The hearts in cntrolgroup were perfused persistently for 100 rain. After being stabilized for 15 min, the I/R group and A-r group hearts were subjected to 40 rain global ischemia by suspension of perfusion followed by 45 rain reperfusion. Coronary effluent was collected continuously for determination of CK activity. At the end of 45 rain repeffusion,the hearts were removed for determination of mycordial SOD activity, MDA content and myocardial infarct size. Cardiomyocyte apoptosis rate, the expression of bcl-2 and bax in hearts were also measured respectively. Results After reperfusion, in A-r group myocardial CK release and MDA content were significantly reduced, the SOD activiy was increasd, the infarct area was significantly lessened, cardiomyocyte apoptosis rate and the expression of bax in hearts were significantly decreased, the expression of bcl-2 in hearts significantly enhanced as compared with I/R group ( all P ＜ 0.01 ). Conclusion Recombinant adeno-associated virus mediated gene transfection of rHO-1 can inhibit the apoptosis in rat cardiomyocytes and enhance the ability of myocardial antioxidation. Gene transfection of rHO-1 has significantly protective effects on isolated myocardial ischemia/reperfusion.