骨髓间质细胞对体外培养的缺氧心肌细胞影响的初步研究

Effect of bone marrow stromal cells on the anoxia cardiomyocytes in vitro

目的:探讨缺氧条件下骨髓间质细胞对培养的心肌细胞凋亡的作用。方法:共培养乳鼠心肌细胞和骨髓间质细胞。利用厌养罐模拟细胞缺氧,观察细胞形态、检测细胞凋亡亚二倍体、观察细胞脱氧核苷酸断裂情况及凋亡相关蛋白的表达。结果:单独培养的骨髓间质细胞对缺氧不敏感,而单独培养的心肌细胞在缺氧后24h出现凋亡,表现为明显的凋亡亚二倍体峰和“DNAladder”。当两种细胞缺氧共培养24h后,混合细胞的凋亡亚二倍体峰明显减少,“DNAladder”消失,抗凋亡蛋白Bcl-2表达无显著差异而促凋亡蛋白Bax表达降低。结论:缺氧条件下体外细胞共培养模型表明骨髓间质细胞对心肌细胞的凋亡有一定的拮抗作用,可能是通过抑制Bax蛋白的表达而实现的。

AIM： To investigate the influence of bone marrow stromal cells （BMSCs） on the anoxia cardiomyocyte apoptosis. METHODS： Using the anaerobic culture apparatus, the apoptosis of the cardiomyocytes, the BMSCs alone and cocultured with each other were detected by morphological observation, PI staining flowcytometry, electrophoretic gel mobility analysis of DNA fragmemation. Western blotting was used to detect Bax and Bcl-2 protein expression. RESULTS：Compared with control, the BMSCs were unsensifive to anode cultured while the anode cardiomyocytes were prone to apoptosis. Apoptosis of cardiomyocytes was increased significandy, detected by PI staining and agarose gel elestrophoresis showed ＂DNA ladder＂. However, when anoxia cardiomyocytes co-cultured with BMSCs, apoptosis cells were decreased, ＂DNA ladder＂ disappeared and the expression of protein Bax was also decreased. CONCLUSION： Bone marrow stromal cells prevent the anoxia cardiomyocytes from apogtosis, probably by suppressing the expgession of Bax protein.