HOE642对大鼠停跳心脏供体心肌细胞凋亡的影响

Effect of HOE642 on Cardiac Myocyte Apoptosis in Rat Non Heart-beating Donors

目的研究HOE642对热缺血死亡大鼠颈部异位停跳心脏供体心肌细胞凋亡的作用。方法取清洁级雄性Sprague-Dawley大鼠112只,随机分成7组,每组16只,各有8只分别作为移植受体和供体。具体如下：C组为正常心脏对照组;S10、S30、S45组分别为心脏停搏后10、30、45 min取心组;SH10、SH30、SH45组分别为心脏停搏后10、30、45 min取心并用HOE642灌注组。对6组实验组大鼠进行热缺血死亡处理,取得供体后,用STH-1液对S10、S30、S45组供体灌注30 min,加20μmol/L的HOE642对SH10、SH30、SH45组供体灌注30 min,再用颈部Cuff法进行异位心脏移植。S10、SH10、S30、SH30组于移植后48 h取供心,S45、SH45组于移植术后取供心。脱氧核糖核苷酸末端转移酶介导的生物素脱氧尿嘧啶核苷酸缺口末端标记法检测供体心肌细胞凋亡,免疫组织化学法检测Bcl-2、Bax、Caspase-3蛋白的表达。结果大鼠腹主动脉横断放血后9-11 min,平均（10.11±0.59）min死亡。S10、S30组的凋亡心肌细胞数显著高于SH10、SH30组（P〈0.05）;S10、S30组的Bcl-2表达水平显著低于SH10、SH30组（P〈0.05）,而Bax、Caspase-3表达水平显著高于SH10、SH30组（P〈0.05）。结论热缺血死亡大鼠颈部异位心脏移植模型是进行停跳心脏供体的心肌保护研究的理想模型;HOE642能抑制热缺血死亡后（30 min内）大鼠心脏心肌细胞凋亡的发生。

Objective To investigate the effect of HOE642 on cardiac myocyte apoptosis of the heterotopic heart transplantation of rat non heart-beating donors.Methods Totally 112 male Sprague-Dawley rats were randomly divided into 7 groups（n=16 in each group）： C,the control group（normal hearts）;S10,S30,and S45（groups of transplanted hearts after 10,30,and 45 minutes of asystole）;and SH10,SH30,and SH45（groups of transplanted hearts after 10,30,and 45 minutes of asystole and infused with HOE642）.After rats in the experimental groups were killed by warm ischemia,the donators of the S10,S30,and S45 groups were infused with STH-1 for 30 minutes,and the dead rats in group SH10,SH30,and SH4 were infused with STH-1 and HOE642（20 μmol/L）for 30 minutes.Heterotopic heart transplantation were processed by the method of neck Cuff.The heart specimens of S10,SH10,S30,and SH30 groups were taken after 48 hours of transplantation,and the heart specimens of S45 and SH45 groups were taken immediately after transplantation.Then apoptotic myocytes were detected with terminal deoxynucleotide transferase-mediated deoxyuridine-biotin nick end labeling method and the expressions of Bcl-2,Bax,and Caspase-3 proteins were detected by immunohistochemistry.Results The rats were discerned death when cardiac electric wave vanished after 9-11 minutes of bloodletting by transsection of abdominal aorta.The number of positive cardiac muscle cells in S10 and S30 groups were significantly larger than those in group SH10 and SH30（P〈0.05）.The levels of Bcl-2 protein expression in S10 and S30 groups were significantly lower than those in SH10 and SH30 groups（P〈0.05）.The levels of Bax and Caspase-3 protein expression were significantly higher than those in SH10 and SH30 groups（P〈0.05）.Conclusions The rat model of a heterotopic heart transplantation on the cervical part is a convenient animal model for cardiac muscle protection.HOE642 can suppress rat cardiac muscle cells apoptosis（within 30 min） after death caused by warm ischemia.