小鼠不同组织及缺氧损伤后的大鼠心肌细胞中RIP3的表达

目的:检测小鼠组织中受体相互作用丝氨酸/苏氨酸蛋白激酶家族(RIPs)表达谱,并检测RIP3在大鼠心肌细胞缺氧损伤后的表达。方法:①采用荧光实时定量PCR分别检测RIPs家族基因在小鼠组织(心、肝、肺、肾、脑、小肠、骨骼肌、脾和主动脉)中的mRNA表达谱,并采用Western blot进一步检测RIP3在小鼠组织的蛋白表达谱。②将培养的大鼠心肌细胞分为缺氧组和对照组,缺氧组置于缺氧环境中培养48 h,采用western blot检测其中RIP3的表达变化。结果:①mRNA水平:RIP1 mRNA在脑组织中表达最高,心脏、肺、肾、骨骼肌较低;RIP2在心脏和肺表达量较其他组织高;RIP3在肠中表达较其他组织高出4倍以上,脑组织中未检测到RIP3表达;RIP4的表达以肺最高,而骨骼肌、脑和血管中表达量低。②蛋白水平:在小鼠组织中,RIP3表达以脑、骨骼肌中最高,心脏、肝、肺中表达较低。③培养的大鼠心肌细胞中,缺氧组心肌细胞的RIP3表达量显著高于对照组(P0.05)。结论:RIPs在小鼠组织中呈现差异表达,而在培养的大鼠心肌细胞缺氧损伤后RIP3表达升高。

Expression Profile of Receptor Interacting Protein 3 in Different MouseTissues and Rat's Myocardial Cells with Hypoxia Injury

Objective： To detect the expression profile of receptor interacting proteins （RIPs） in mouse tissues, and to detect the expression of RIP3 in rat＇s myocardial cells with hypoxia injury. Methods：① We detected the expression profile of RIPs family in mouse tissues （heart, liver, lung, kidney, brain, intestine, skeletal muscles, spleen and aorta） by real-time fluorescent quantitative PCR, then detected the expression profile of RIP3 protein. ② The myocardial cells of rat were divided into hypoxia group and control. We treated the first group for 48 h on hypoxia to obtain hypoxia injured models, then quantitated the expression of RIP3 by western blot. Results：③ On mRNA level, the expression of RIP1 was highest in brain, but lower in heart, lung, kidney and skeletal muscle; the expression of RIP2 was higher in heart and lung than other tissues; RIP3 in the intestine was 4 times higher than other, but it was very low expression in brain; the expression of RIP4 was highest in lung, and was lowest in skeletal muscle, brain, spleen, and vessel. ④ Western blot results showed the expression of RIP3 at protein level. It was higher expressed in brain, skeletal muscle, however, lower in heart, liver and lung. ⑤After treated on hypoxia, RIP3 of hypoxia group was significantly higher than control in the cultured rat myocardial cell（P〈0.05）. Conclusions： The expression of RIPs represented various level in mouse tissues. RIP3 increased in rat myocardial cells under hypoxia injured condition.