论文部分内容阅读
该文主要探讨Akt/HIF-1α(hypoxia inducible factor-1α)信号通路在二氧化硒(Se O2)诱导大鼠肾上腺嗜铬细胞瘤PC12细胞损伤中的作用。将PC12细胞暴露于不同浓度的Se O2(40、80、160μmol/L)24 h以诱导细胞发生损伤。采用噻唑蓝还原法和乳酸脱氢酶漏出率检测法测定细胞损伤程度,倒置显微镜观察细胞形态的变化,用丙二醛(malonic dialdehyde,MDA)和超氧化物歧化酶(superoxide dismutase,SOD)试剂盒检测细胞内活性氧类活性氧类(reactive oxygen species,ROS)水平,Hoechst 33342单荧光染色法观察细胞凋亡,免疫印迹法检测细胞HIF-1α、磷酸化Akt(phosphorylated Akt,p-Akt)、淋巴瘤/白血病-2(B cell lymphoma/leukemia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、PI3k、p53和Caspase-3(cysteinyl aspartate specific proteinase-3)的表达。结果显示,二氧化硒可呈剂量依赖性地诱导PC12细胞损伤,导致细胞内ROS增多和细胞凋亡,引起细胞皱缩,轴突变短。p-Akt、HIF-1α、p53、Caspase-3表达上调,Bax/Bcl-2表达比例显著增加。由此说明,二氧化硒诱导PC12细胞损伤,导致细胞凋亡,与其激活细胞Akt/HIF-1α信号通路,进而促进p53、Bax/Bcl-2、Caspase-3的表达及胞内ROS增加有关。
The aim of this study is to investigate the role of Akt / HIF-1α signaling pathway in selenium dioxide (Se O2) -induced PC12 cell injury in rat adrenal pheochromocytoma. PC12 cells were exposed to different concentrations of Se O2 (40,80,160 μmol / L) for 24 h to induce cell injury. The degree of cell damage was determined by thiazolyl blue reduction method and lactate dehydrogenase leakage rate test. The morphological changes of cells were observed by inverted microscope. Malondialdehyde (MDA) and superoxide dismutase (SOD) The levels of reactive oxygen species (ROS) in cells were detected by flow cytometry. The apoptosis of HIF-1α, phosphorylated Akt (p-Akt) cells was detected by Hoechst 33342 single fluorescent staining. , B cell lymphoma / leukemia-2 (Bcl-2), Bcl-2 associated X protein (Bax), PI3k, p53 and Caspase-3 (cysteinyl aspartate specific proteinase -3) expression. The results showed that selenium dioxide induced PC12 cell damage in a dose-dependent manner, resulting in increased intracellular ROS and apoptosis, causing cell shrinkage and shortening of axons. The expressions of p-Akt, HIF-1α, p53 and Caspase-3 were up-regulated and the ratio of Bax / Bcl-2 expression was significantly increased. Thus, selenium dioxide induced PC12 cell injury, leading to apoptosis, which was related to its activation of Akt / HIF-1α signaling pathway, and then to the expression of p53, Bax / Bcl-2 and Caspase-3 and the increase of intracellular ROS.