论文部分内容阅读
目的研究在缺氧/复氧条件下随着缺氧时间延长αB-晶体蛋白对原代培养心肌细胞中caspase-3的影响。方法以阳离子脂质体(LipofectamineTM2000)介导,将原代培养SD大鼠乳鼠心肌细胞分为转染组(转染pEGFP-αB-crystallin-C3)、空载组(转染pEGFP-C3)和空白组(无任何质粒转染),转染后分别缺氧培养6、12、18h后复氧12h,Western Blot半定量检测αB-晶体蛋白和caspase-3活化产物p17表达。结果转染组心肌细胞αB-晶体蛋白表达有下降趋势,但差异无显著性(P>0.05);转染组p17与空白组和空载组比较,明显降低(P<0.05),在缺氧6～18h内,随着缺氧时间延长,表达相对稳定,有下降趋势但差异无显著性(P>0.05);在缺氧12h后,各组p17表达量趋于平稳。结论在缺氧/复氧条件下,αB-晶体蛋白可能具有抑制caspase-3活化,抗细胞凋亡作用;在缺氧6～18h内,随着缺氧时间延长,αB-晶体蛋白抑制caspase-3活化的作用虽有下降趋势,但基本相对稳定。
Objective To investigate the effects of αB-crystallin on caspase-3 in cultured primary cultured cardiomyocytes under hypoxia / reoxygenation condition. Methods Primary cultured SD rat neonatal rat cardiomyocytes were transfected with pEGFP-αB-crystallin-C3 and non-transfected with pEGFP-C3 by LipofectamineTM2000. And blank group (without any plasmid transfection). After transfection, cells were reoxygenated for 6h, 12h and 12h after transfection respectively. Western Blot was used to detect the expression of αB-crystallin and caspase-3 activation product p17. Results The expression of αB-crystallin in cardiomyocytes of transfected group showed a decreasing trend, but the difference was not significant (P> 0.05). Compared with the blank group and the no-load group, the expression of p17 in the transfection group was significantly lower (P <0.05) Within 6 to 18 hours, the expression of p17 was relatively stable with a decreasing trend but no significant difference (P> 0.05). The expression of p17 in each group tended to be stable after hypoxia for 12 hours. Conclusion Under hypoxia / reoxygenation conditions, αB-crystallin may inhibit caspase-3 activation and anti-apoptotic effect. Within 6-18 h of hypoxia, αB-crystallin inhibits caspase- 3 Although the role of activation of the downward trend, but basically relatively stable.