类糖尿病环境对P19细胞分化为心肌细胞的影响  

The effects of diabetes-like environment on P19 cell for myocardial cell differentiation

作　　者：方旭[1] 杨荣[1] 徐春阳[1] 张浩[1] 钱玲梅[1] 盛燕辉[1] 孔祥清[1] 

机构地区：[1]南京医科大学第一附属医院心血管科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2013年第5期621-625,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然基金青年基金项目(30901473);江苏省卫生厅科教兴卫工程重点学科开放课题(ZX07200905)

摘　　要：目的:探讨在类糖尿病环境下诱导P19细胞向心肌细胞分化过程中心肌转录因子GATA结合蛋白4(GATA bindingprotein 4,GATA4)和心肌特异标志物心肌肌钙蛋白T(cardiac troponin T,cTnT)的表达水平。方法:体外类糖尿病环境下,诱导P19细胞向心肌细胞分化,通过Real-time PCR和蛋白质印迹法分别检测其cTnT和GATA4在不同时间点的mRNA以及蛋白表达水平。结果:在分化过程中,类糖尿病组P19细胞分化形成的胚胎样小体较正常组增殖缓慢,形成的生长晕松散零乱。类糖尿病环境下GATA4 mRNA的表达量在分化第6、8、10天(0.05±0.06、0.11±0.04、0.45±0.12)均明显低于同时间点的对照组(1.00±0.04、0.44±0.06、0.78±0.20),cTnT mRNA的表达量(0.12±0.03、0.07±0.04、0.46±0.11)均明显低于同时间点的对照组(1.02±0.25、0.25±0.02、0.71±0.21)。蛋白质印迹法结果用积分光密度值量化数据,类糖尿病环境下分化第6、8、10天GATA4蛋白表达(0.40±0.06、0.25±0.03、0.92±0.13)均明显低于同时间点的对照组(0.69±0.09、0.75±0.08、1.05±0.07)。类糖尿病环境下分化第6、8天cTnT蛋白表达(0.39±0.08、0.37±0.05)均明显低于同时间点的对照组(0.79±0.05、0.54±0.07)。统计结果表明在分化第6、8天时,类糖尿病环境下GATA4和cTnT mRNA和蛋白的表达量与对照组相比,差异均有统计学意义。结论:类糖尿病环境培养降低了P19细胞向心肌细胞的分化。Objective：To investigate myocardial transcription factor GATA4 and myocardial specific marker cardiac tropenin T （cTnT） expression of P19 cells differentiated into cardiomyocytes in diabetes-like environment, nethods：P19 cells were induced to myocardial cells in supplemented diabetic teratogenic medium. Real-time PCR and Western blot were performed to detect GATA4 and cTnT expressions in the two groups on differentiated process. Results： Compared with the control group,the elementary bodies（EB） that cultured in diabetes-like environment propagated obviously slower and the aggregation outgrowths were scattering and sparse. The volumes of GATA4 in diabetes-like environment medium on day 6,8 and 10（0.05 ±0.06,0.11 ± 0.04 and 0.45 ± 0.12） of culture were significantly lower than the same time of the control group （1.00 ± 0.04,0.44 ± 0.06 and 0.78 ± 0.20） of culture respectively. The volumes of cTnT in diabetes-like environment medium on day 6,8 and 10 （0.12 ± 0.03,0.07 ± 0.04 and 0.46 ± 0.11 ） of culture were significantly lower than the same time of the control group （1.02 ± 0.25,0.25 ± 0.02 and 0.71 ± 0.21） of culture respectively. Western blot assay resulted with the integral optical density （IOD） values of quantitative data. The protein expressions of GATA4 in diabetes-like environment medium on day 6,8 and 10 （0.40 ± 0.06,0.25 ± 0.03 and 0.92 ± 0.13） of culture were significantly lower than the same time of the control group （0.69 ± 0.09,0.75 ±0.08 and 1.05 ±0.07）. The protein expressions of cTnT in diabetes-like environment medium on day 6 and 8 （0.39 ± 0.08,0.37 ±0.05） of culture were significantly lower than the same time of the control group （0.79 ± 0.05,0.54 ± 0.07）. There were significant differences between the day 6 and 8 groups （P 〈 0.05）. Conclusion：P19 cell induced decreasingly to myocardial cell differentiation in diabetes-like environment culture.

关 键 词：类糖尿病环境 P19细胞 分化 心肌细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]