摘要
目的 研究蛋白激酶C信号通路在心肌细胞缺氧性凋亡中的作用。方法 将培养的大鼠心肌细胞随机分成 8组 :(1 )对照组 ;(2 )缺氧 6h组 ;(3)缺氧 1 2h组 ;(4 )缺氧 2 4h组 ;(5 )PMA1 0nM组 ;(6 )PMA1 0 0nM组 ;(7)CHE1 μM组 ;(8)CHE1mM组。对照组常氧培养 2 4h。缺氧培养即将心肌细胞 95 %N2 ～ 5 %CO2 下 37℃缺氧培养。 (5 )和 (6 )组即于缺氧培养前 ,加入PKC激动剂PMA(Phorbol 1 2 myristate 1 3 acetata) ,使其终浓度分别为 1 0nM、1 0 0nM ,预处理 1 0min后再缺氧 2 4h。 (7)和 (8)组即于缺氧培养前 1 0min ,加入PKC抑制剂CHE(chelerythrine) ,使其终浓度分别为 1 μM、1mM ,然后再缺氧培养 2 4h。其余各组缺氧不同的时间。终止培养后分别检测细胞的凋亡率和活性。结果 单纯缺氧能引起心肌细胞的凋亡和坏死 ,以缺氧 2 4h最显著。PMA1 0nM预处理能明显减少心肌细胞缺氧性凋亡 (P <0 .0 5 ) ,而PMA1 0 0nM组和CHE1 μM组心肌细胞凋亡率明显增加 (P <0 .0 5 )。PMA和CHE对 2 4h缺氧诱导的心肌细胞坏死无明显影响。结论 蛋白激酶C信号传导通路参与了抗心肌细胞缺氧性凋亡的信号传导 。
Objective To investigate the roles of protein kinase C signal transduction in apoptosis of cardiomyocytes induced by hypoxia.Methods Cultured neonatal rat cardiomyocytes were randomly assigned to eight groups including control group, hypoxia for 6h group, hypoxia for 12 h group, hypoxia for 24 h group, PMA(Phorbol 12 myristate 13 acetata)10nM group, PMA 100nM group, CHE(chelerythrine)1μM group and CHE1mM group. In control group, cardiomyocytes were cultured under 95%O 2 and 5% CO 2;In PMA 10nM group and PMA 100nM group, cardiomyocytes were cultured for 24 h under 95%N 2 and 5% CO 2 after being preconditioned by PMA10nM or PMA 100nM respectively; cardiomyocytes were cultured for 24 h under 95%N 2 and 5% CO 2 after being preconditioned by CHE1μM or CHE1mM respectively in CHE 1μM group and CHE 1mM group. After culture was ended, apoptosis cells were measured by TUNEL label in individual cardiomyocyte, and viability was determined by trypan blue exclusion. Results Both apoptosis and necrosis were induced by hypoxia in cultured rat cardiomyocytes with a time dependent manner. Less apoptosis cells were induced by hypoxia in PMA 10nM group than in hypoxia for 24 h group( P <0.05), but in comparison with hypoxia for 24 h group, more apoptosis cells were induced in PMA 100nM group and CHE1μM group( P <0.05).Apart from control group, the cell viabilities of the other groups were not statistically significant. Conclusion Protein kinase C is involved in anti apoptosis signal in cardiomyocytes during hypoxia. The anti apoptosis signal and anti necrosis signal of PKC might be distinct during hypoxia.
出处
《重庆医学》
CAS
CSCD
2003年第8期973-974,986,共3页
Chongqing medicine
关键词
心肌细胞
蛋白激酶C
凋亡
缺氧
信号传导
cardiomyocytes
protein kinase C
apoptosis
signal transduction
hypoxia
