论文部分内容阅读
【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 investigate whether transfection of human anti-apoptotic gene bcl-xL can improve cardiomyocyte hypoxia tolerance and evaluate its anti-apoptotic effect on cardiomyocytes. 【Methods】 Cardiomyocytes were cultured in vitro to establish simulated myocardial hypoxia model. Cardiomyocytes were infected with recombinant adenovirus carrying human bcl-xL gene. The expression of bcl-xL protein was analyzed by immunocytochemical staining. TUNEL assay The apoptosis index was detected. The green fluorescent protein was observed under the fluorescence microscope and the morphological changes of the nucleus were observed by Hoechst33342 staining. 【Results】 The positive expression of bcl-xL protein (17.84 ± 3.42) in cardiomyocytes transfected with rAd-bcl-xL / s was significantly higher than that of rAd-bcl-xL / The apoptotic index of cardiomyocytes after hypoxia induction was (45.67 ± 8.47)%, while the apoptosis index of transfected rAd-bcl-xL / s decreased to (9.42 ± 2.25) (P <0.01) ± 2.48)%, there was a significant difference (P <0.01), no typical apoptotic bodies. 【Conclusion】 Adenovirus-mediated bcl-xL gene can efficiently transfect cardiomyocytes and express the target gene; exogenous bcl-xL gene has anti-apoptotic effect on cultured cardiomyocytes, Tolerance, promote the recovery of injured myocardium.