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该文研究了体外培养肝细胞内钙离子浓度改变对细胞存活率、凋亡和增殖的影响。建立了H2O2诱导小鼠胚胎肝细胞损伤模型,CCK-8检测细胞存活率,Fura-2/AM负载检测细胞内[Ca2+]i;免疫荧光和Western blot分别检测STIM1和Orai1在细胞内的定位和含量;流式细胞术检测细胞凋亡;Brdu掺入检测细胞增殖。结果显示,H2O2刺激后细胞存活率降低为对照组的73%,凋亡细胞比例增加,增殖细胞数目显著减少,细胞内[Ca2+]i升高,STIM1和Orai1蛋白质水平增加,且STIM1可与Orai1蛋白质共定位。2-APB预处理组可以降低细胞内[Ca2+]i,减少STIM1和Orai1蛋白质表达水平,抑制STIM1和Orai1蛋白质的相互作用。结果表明,H2O2可通过影响细胞内钙离子稳态导致细胞凋亡。
This article studies the effects of changes of intracellular calcium concentration on cell viability, apoptosis and proliferation in vitro. H2O2-induced mouse embryonic hepatocyte injury model was established. Cell viability was detected by CCK-8, and [Ca2 +] i was detected by Fura-2 / AM. The intracellular localization of STIM1 and Orai1 were detected by immunofluorescence and Western blot, respectively Content; apoptosis was detected by flow cytometry; Brdu incorporation was used to detect cell proliferation. The results showed that after H2O2 stimulation, the cell viability decreased to 73% of the control group, the percentage of apoptotic cells increased, the number of proliferating cells decreased significantly, [Ca2 +] i increased, the protein levels of STIM1 and Orai1 increased, and STIM1 and Orai1 Protein co-localization. The 2-APB preconditioning group decreased [Ca2 +] i, decreased the expression of STIM1 and Orai1 protein, and inhibited the interaction of STIM1 and Orai1 protein. The results showed that H2O2 can induce cell apoptosis by affecting intracellular calcium homeostasis.