摘要
目的:研究MAPK通路在原癌基因Pim-3抗心肌急性缺氧复氧损伤中的作用。方法:采用原代培养新生大鼠的心肌细胞,随机分为4组:正常对照组(control)、缺氧复氧组(A/R)、缺氧预适应组(APC+A/R)、阻断剂组。在缺氧预处理前分别用终浓度为10μmol/LSB203850(p38MAPK阻断剂)、U0126(ERK1/2阻断剂)、SP600125(SAPK/JNK阻断剂)与细胞孵育30min。实验结束后测定MAPKs通路中ERK1/2、JNK、p38MAPK磷酸化蛋白表达水平及Pim-3蛋白的表达水平,同时检测培养液中乳酸脱氢酶(LDH)活性、四唑盐(MTT)比色试验测定细胞存活率、TUNEL法检测细胞凋亡。结果:SB203850、U0126、SP600125能分别取消由APC或A/R所诱导ERK1/2、JNK、p38MAPK的磷酸化水平的升高;由APC所诱导的Pim-3表达的升高在p38MAPK通路被阻断后明显下调(P<0.01),并且心肌细胞LDH值升高,细胞存活率则下降,心肌细胞的凋亡指数升高。结论:p38MAPK的激活可上调原癌基因Pim-3的表达,从而可能对心肌细胞起到保护作用。
AIM： To investigate the role of mitogen- activated protein kinases （MAPKs） pathways and the molecular mechanism by which the proto - oncogene Pim - 3 protects cardiomyocyte against anoxia/reoxygenation （A/R） injury. METHODS： The primarily cultured neonatal rat ventricular cardiomyocytes were randomly divided into 4 groups： control group; A/R group; APC ＋ AiR group; SB203850, U0126 or SP600125 ＋ APC ＋ A/R group. The cells were pre - incubated with U0126 （ERK1/2 inhibitor）, SP600125 （SAPK/JNK inhibitor）, or SB203850 （p38 MAPK inhibitor） at concentration of 10 μmol/L for 30 rain before the APC. The activities of p38 MAPK, JNK and ERK1/2 were detected by Western blotting. The viability of cardiomyocytes was assayed by MTT and the apoptosis of cardiomyocyte was detected by TUNEL. RESULTS： U0126, SB203850, and SP600125 abolished the increased expression of ERK1/2, p38 -MAPK, and JNK proteins induced by APC ＋ A/R or A/R, respectively. The expression level of Pim - 3 protein significantly decreased when the p38 MAPK signal pathway was inhibited. Meanwhile, the activity of LDH and the apoptosis index increased, and the viability of cardiomyocytes decreased. CONCLUSION： Pim- 3 expression through a p38 MAPK signaling pathway may protect cardiomyocytes from A/R injury.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第10期1912-1916,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30860271)
