高糖对乳鼠心肌细胞凋亡及相关基因表达的影响

Effects of high glucose on apoptosis and expression of apoptosis-related genes in neonatal rat cardiomyocytes

目的:探讨高糖对乳鼠心肌细胞凋亡及相关基因表达的影响。方法:分离培养SD乳鼠心肌细胞,以20mmol/L终浓度的葡萄糖干预48h后,激光共聚焦显微镜、TUNEL染色和流式细胞术检测心肌细胞凋亡;荧光定量PCR检测高糖对凋亡相关基因P53、Pax3、Bmp4、Slug、Msx1表达的影响。结果:激光共聚焦显微镜观察可见高糖组心肌细胞凋亡明显增加;TUNEL检测心肌细胞凋亡指数,高糖组为(19.67±3.45)%,明显高于对照组(1.12±0.24)%(P<0.05);流式细胞术显示高糖组细胞凋亡率较对照组明显增加(P<0.05);荧光定量PCR表明高糖能上调P53、Msx1基因的表达,下调Pax3、Bmp4、Slug基因的表达。结论:高糖可能通过影响凋亡相关基因P53、Pax3、Bmp4、Slug、Msx1的表达而诱导心肌细胞凋亡。

Objective：To investigate the effects of high glucose on apoptosis and expression of apoptosis-related genes in neonatal rat cardiomyocytes. Methods：Cultured cardiomyocytes were exposed to high glucose （20 mmol/L） for 48 h. Laser confocal microscopy, TUNEL assay and flow cytometry were utilized to detect cardiomyocyte apoptosis. Real-time PCR was performed to identify mRNA expression of apoptosis-related P53,Pax3, Bmp4,Slug and Msx1 genes in the glucose group and the control group. Results：Cardiomyocyte apoptosis was obviously observed in the glucose group under the laser confocal microscope. TUNEL assay indicated that cardiomyocyte apoptotic index in the glucose group was significantly higher than that in the control group （P 〈 0.05）. Flow cytometry revealed that cardiomyocyte apoptotic rate in the glucose group was significantly higher than that in the control group （P 〈 0.05）. Real-time PCR Showed that the expressions of P53 and Msx were up-regulated in the glucose group while the expressions of Pax3, Bmp4 and Slug were down-regulated. Conclusion：High glucose can disrupt apoptosis-related genes’ expression, including P53,Pax3, Bmp4,Slug and Msx1, which may be related to the cardiomyocyte apoptosis.