摘要
目的探讨Bcl-2基因转染在小鼠心脏移植急性排斥反应模型中对心肌细胞凋亡的影响和作用机制。方法通过TDT介导的原位末端标志(TUNEL)、DNA阶梯及Western免疫印迹(WesternBlot)等检测方法,分析了小鼠心脏移植模型急性排斥反应时细胞凋亡的情况,并结合心脏移植急性排斥反应的组织学分级,探讨了心肌细胞凋亡在急性排斥反应中的作用。同时应用免疫组织化学染色(SABC法)分析活化型Caspase3蛋白的表达,探讨了Bcl2基因转染对心肌细胞凋亡的影响及可能作用机制。结果与同种异体移植组比较,Bcl2基因转染后能显著减少急性排斥反应中的心肌细胞凋亡现象,同时,活化型Caspase3蛋白的表达从9.75±2.71被下调到3.56±0.62(P<0.01)。结论小鼠心脏移植急性排斥反应过程中存在心肌细胞的凋亡。它是小鼠心脏移植急性排斥反应中心肌组织损伤的主要机制,Bcl2基因转染能有效地抑制心肌细胞的凋亡,可能主要通过阻断细胞凋亡信号传导途径的中下游阶段,达到阻止Caspase3蛋白酶活化而发挥抗心肌凋亡作用。
Objective To investigate the effect and action mechanis ms of Bcl-2 gene transfer on apoptosis of myocardial cells in mice heart transpla ntation.Methods Measures including TUNEL,DNA ladders that wer e showed by electrophoresis on agarose gels and Western blot were used to analyz e the apoptosis of allograft acute rejection after the mice heart transplantatio n.With histological studies,we investigated the roles of apoptotic cells in th e cardiac allograft rejection.Also by using immunohistochemistry method to stud y the expression of active form of Caspase 3 protease,we try to find the effect and action mechanisms of Bcl-2 gene transfer on apoptosis of myocardial cells. Results Compared with allograft group,Bcl-2 gene transfer g roup decrease d significantly the apoptosis of myocardial cell in acute rejectio n.Meanwhile the expression of active form of Caspase 3 protease was cu t down from(9.75±2.71) to (3.56±0.62)(P<0.01). Con clusions Apoptosis of myocardial cells occurrs in acute rejection afte r mice heart transplantation.It is the main mechanisms of damage of myocardial tissue for the acute rejection after mice heart transplantation.Bcl-2 gene tra nsfer can efficiently choke back the apoptosis of myocardial cells.Bcl-2 may b e cut-out the channel of information transmission of apoptosis in middle-late stage to prevent Caspase 3 protease from activation so that the anti-apoptosis of myocardial cells will be achieved.
出处
《广西医学》
CAS
2005年第5期629-633,共5页
Guangxi Medical Journal
基金
德国Essem大学医学院资助项目
德国Aachem理工大学医学院资助项目
