重组bcl-xL腺病毒对心肌细胞凋亡的影响  

Effect of Recombinant Adenovirus Containing bcl-xL Gene on Apoptosis of Cardiac Myocytes

作　　者：廖洪映[1] 熊利华[1] 陈波[1] 张惠忠[1] 华平[1] 殷香保[2] 祝曙光[1] 傅玉如[2] 

机构地区：[1]中山大学附属第二医院心胸外科,广东广州510120 [2]中山大学附属第二医院医学研究中心,广东广州510120

出　　处：《中山大学学报（医学科学版）》2005年第4期421-423,427,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：卫生部科研基金资助项目(981093)

摘　　要：【目的】探讨转染人类抗凋亡基因bcl-xL能否提高心肌细胞对缺氧的耐受性,评价它对心肌细胞的抗凋亡作用。【方法】体外培养SD大鼠乳鼠心肌细胞,建立模拟心肌缺氧模型;将携带人类bcl-xL基因的重组腺病毒感染心肌细胞;免疫细胞化学染色法分析bcl-xL蛋白的表达;TUNEL法检测凋亡指数;荧光显微镜下观察心肌细胞表达绿色荧光蛋白,用Hoechst33342染色观察细胞核的形态变化。【结果】转染rAd-bcl-xL/s的心肌细胞表达bcl-xL蛋白阳性单位(PU)(17.84±3.42)明显高于正常对照组(11.02±2.56)与rAd-bcl-xL/as转染组(8.53±2.25)(P<0.01);缺氧诱导后心肌细胞凋亡指数为(45.67±8.47)%,而转染rAd-bcl-xL/s后的细胞凋亡指数减少至(9.42±2.48)%,有显著性差异(P<0.01),未见典型的凋亡小体。【结论】腺病毒介导的bcl-xL基因能高效转染心肌细胞,并表达目的基因;外源性bcl-xL基因对体外培养的心肌细胞具有抗凋亡作用,能够提高心肌细胞对缺氧的耐受性,促进损伤心肌的恢复。[Objective] To study the effects of bcl-xL gene on the apoptosis of cardiac myocytes after hypoxia in vitro. [Methods] Cardiac myocytes of neonatal rat were cultured and infected by recombinant adenovirus containing bcl-xL gene, then cultured under hypoxic condition. The bcl-xL protein expression was detected by immunochemistry. The apoptosis index was measured by TUNEL; and the apoptosis bodies were observed by Hoechst33342 staining under fluorescence microscope. [Results]The expression of bcl-xL protein (Positive unit,PU)was increased to (17.84±3.42) PU. The apoptosis index of cardiac myocytes after hypoxic was (45.67±8.47)%, then it decreased to (9.42±2.48)% by rAd-bcl-xL/s; No typical apoptosis body was detected in rAd-bcl-xL/s group. [Conclusions] Recombinant adenovirus containing bcl-xL gene can transfect cardiac myocytes efficiently and the exogenous bcl-xL gene expressed highly in vitro. bcl-xL gene transfection can reduce apoptosis of cardiac myocyte. bcl-xL may serve as a potential therapeutic tool for cardioprotection.

关 键 词：心肌细胞凋亡 BCL-XL基因 HOECHST33342 重组 免疫细胞化学染色法 心肌细胞表达 细胞凋亡指数 抗凋亡作用 TUNEL法 绿色荧光蛋白 显微镜下观察 体外培养 抗凋亡基因 腺病毒感染 正常对照组 显著性差异 腺病毒介导 SD大鼠 

分 类 号：R542.2[医药卫生—心血管疾病] R456[医药卫生—内科学]