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目的:研究KATP通道开放剂埃他卡林在缺氧诱导的肺血管内皮细胞凋亡调节中的作用及机制。方法:细胞分组培养并经过药物及缺氧处理后,分别采用四甲基偶氮唑盐(MTT)法对细胞活性进行测定,同时采用Hoechst 33342染色法检测细胞凋亡,采用Western blot对JNK、磷酸化JNK、caspase-3等蛋白的表达进行检测。结果:缺氧可降低细胞活性,诱导肺动脉内皮细胞凋亡,而埃他卡林可抑制凋亡,这种细胞保护作用可被KATP拮抗剂5-HD阻断;埃他卡林可抑制缺氧引起的c-JNK通路的磷酸化激活,并抑制下游凋亡蛋白caspase-3的表达,与c-JNK抑制剂SP600125作用一致,而5-HD处理组的趋势则相反。结论:埃他卡林可通过开放KATP通道抑制缺氧诱导的肺动脉内皮细胞凋亡,这种保护机制涉及了对c-JNK通路及下游凋亡蛋白的调节。
OBJECTIVE: To study the role and mechanism of ethakalim, a KATP channel opener, in the regulation of hypoxia-induced pulmonary vascular endothelial cell apoptosis. Methods: The cells were cultured in groups and treated with hypoxia. MTT assay was used to determine the cell viability. At the same time, cell apoptosis was detected by Hoechst 33342 staining. The expression of JNK, Phosphorylation of JNK, caspase-3 and other proteins were detected. RESULTS: Hypoxia decreased cell viability and induced apoptosis of pulmonary artery endothelial cells, whereas iptakalim inhibited apoptosis. This cell-protective effect was blocked by 5-HD, a KATP antagonist. Oxytocin inhibited hypoxia Induced the phosphorylation of c-JNK pathway and inhibited the expression of caspase-3, a downstream inhibitor of apoptosis, which was consistent with the effect of c-JNK inhibitor SP600125, while the trend of 5-HD treatment was opposite. CONCLUSION: Iptakalim inhibits hypoxia-induced apoptosis of pulmonary artery endothelial cells by opening KATP channels. This protective mechanism involves the regulation of c-JNK pathway and downstream apoptotic proteins.