体外心肌细胞缺血后适应模型的建立及评价

目的建立心肌细胞缺血后适应（IPost）模型并观察IPost对心肌细胞再灌注（I／R）损伤的影响。方法体外培养新生SD乳鼠心肌细胞，随机分正常对照组（SH组）、缺血再灌注组（I／R组）及缺血后适应组（IPost组），用烟酸己可碱（Hochest33342）及碘化丙啶混合溶液染色后观察细胞凋亡率情况，流式细胞术检测细胞凋亡率，逆转录PCR（RT—PCR）方法和蛋白质印迹法（WesternBlot）方法检测Bcl-2／Bax基因表达。结果与I／R组比较，IPost组细胞凋亡率显著改善，Bcl-2蛋白表达上调，Bax蛋白表达下降，Bcl-2／Bax显著增加（P均d0．05）。结论IPost能通过抗心肌凋亡有效地减轻离体心肌细胞缺血再灌注损伤。

The establishment and evaluation of vitro ischemic postconditioning modle of myocardial cells

Objective To evaluate the effect of ischemic postconditioning （IPost） on ischemia reperfusion injury by establishing the Ipost modle of myocardial cells. Methods Myocardial cells of SD neonatals were cultured in vitro, and divided randomly into 3 groups： SH group, I/R group and IPost group. The morphology of myocardial cells was cultured through inverted fluorescence microscope. The cells were stained by Hochest33342 to observe the cell＇s apoptosis situation. Flow cytometry was adopted to evaluate the apoptosis rate of cells; RT-PCR and Western Blot were used to test the mRNA of Bcl-2/Bax. Results Compared with the I/R group, the apoptosis rate in IPost group was greatly improved, the expression of Bcl-2 was upregulated while Bax downregulated, and Bcl 2/Bax increased （P 〈0.05）. Conclusion IPost can protect the myocardiol cells in vivo from ischemia reperfusion injury by opposing to the apoptosis of myocardiol cells.