文章摘要
方 旭,杨 荣,徐春阳,张 浩,钱玲梅,盛燕辉,孔祥清.类糖尿病环境对P19细胞分化为心肌细胞的影响[J].南京医科大学学报,2013,(5):621~625
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
投稿时间:2012-12-19  
DOI:10.7655/NYDXBNS20130511
中文关键词: 类糖尿病环境  P19细胞  分化  心肌细胞
英文关键词: diabetes-like environment  P19 cell  differentiation  myocardial cell
基金项目: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)
作者单位
方 旭 南京医科大学第一附属医院心血管科,江苏 南京 210029 
杨 荣 南京医科大学第一附属医院心血管科,江苏 南京 210029 
徐春阳 南京医科大学第一附属医院心血管科,江苏 南京 210029 
张 浩 南京医科大学第一附属医院心血管科,江苏 南京 210029 
钱玲梅 南京医科大学第一附属医院心血管科,江苏 南京 210029 
盛燕辉 南京医科大学第一附属医院心血管科,江苏 南京 210029 
孔祥清 南京医科大学第一附属医院心血管科,江苏 南京 210029 
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中文摘要:
      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 binding protein 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。
英文摘要:
      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. Methods: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.
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