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目的研究硒对于H2O2诱导大鼠肝细胞(BRL)凋亡的影响并探讨其作用机制。方法采用0.1mmol/L的H2O2建立氧化损伤的细胞模型,并施加不同剂量的补硒干预。通过测定细胞活力(MTT)、谷胱甘肽过氧化物酶(GPH-Px)活力、丙二醛(MDA)含量和细胞凋亡率,研究硒对氧化损伤肝细胞的保护作用,采用实时定量PCR技术检测细胞GPH-Px和凋亡信号调节激酶1(ASK1)的mRNA表达水平,并通过Western blot技术检测ASK1的蛋白表达水平。结果补硒组细胞MTT(OD)值和GSH-Px活力高于损伤组,MDA的含量和细胞凋亡率低于损伤组,差别有统计学意义(P<0.05)。补硒组GPH-Px的mRNA表达水平高于损伤组,差别有统计学意义(P<0.05),补硒组ASK1的mRNA和蛋白表达水平低于损伤组,差别有统计学意义(P<0.05)。结论硒可能通过增加细胞GSH-Px的mRNA表达和酶活性、下调ASK1 mRNA和蛋白的表达、降低细胞中MDA含量等作用,对氧化损伤所致的肝细胞凋亡起到一定的保护作用。[营养学报,2010,32(5):466-469]
Objective To study the effect of selenium on H2O2-induced apoptosis in rat hepatocytes (BRL) and to explore its mechanism. Methods 0.1 mmol / L H2O2 was used to establish a cell model of oxidative damage and intervention with different dosages of selenium was performed. The protective effect of selenium on oxidative damage of hepatocytes was studied by measuring cell viability (MTT), glutathione peroxidase (GPH-Px) activity, malondialdehyde (MDA) content and apoptosis rate. Quantitative real- PCR was used to detect the mRNA expression of GPH-Px and ASK1. The protein expression of ASK1 was detected by Western blot. Results The MTT (OD) value and GSH-Px activity in selenium-supplemented group were higher than those in injured group. The content of MDA and the apoptosis rate of selenium-deficient group were lower than those of injury group (P <0.05). The mRNA expression of GPH-Px in the selenium-supplemented group was higher than that in the injury group (P <0.05), and the mRNA and protein expression of ASK1 in selenium-supplemented group was lower than that in the injury group (P <0.05) ). Conclusion Selenium may play a protective role in hepatocellular apoptosis induced by oxidative damage by increasing the mRNA and protein expression of GSH-Px, decreasing the expression of ASK1 mRNA and protein, and decreasing the content of MDA in cells. [Journal of Nutrition, 2010,32 (5): 466-469]