Flotillin-2在高氧诱导肺上皮细胞凋亡中的作用

发布时间：2018-06-06 17:30

本文选题：Flotillin- + 肺上皮细胞　；参考：《广东医学》2017年05期

【摘要】：目的了解Flotillin-2(Flot-2)在高氧诱导肺上皮细胞凋亡中的调控分子机制。方法把小鼠置于高氧环境中处理24 h后,取其肺脏切片通过免疫组化检测Flot-2的表达。通过体外肺上皮细胞株进一步验证高氧后Flot-2在细胞中基因和蛋白水平的表达情况。通过质粒转染的方式把Flot-2的基因整合进肺上皮细胞的基因组中使Flot-2高表达,把肺上皮细胞分别置于空气和高氧环境中继续培养24 h,收集细胞后提取总蛋白,通过Westen blot检测经典凋亡途径中的各个相关因子,包括Caspase-3、Caspase-9,以及线粒体和细胞质中的细胞色素C,进一步研究在Flot-2高表达情况下,高氧对细胞凋亡的影响。结果 Flot-2广泛存在于支气管上皮和肺泡上皮细胞中,高氧刺激后表达明显增加。肺上皮细胞经质粒转染高表达Flot-2之后接受高氧刺激,细胞的死亡减少,凋亡途径中关键的凋亡蛋白Caspase-3及其活性片段cleaved-Caspase-3明显减少。内源性线粒体途径中关键的凋亡蛋白Caspase-9及其活性片段cleaved-Caspase-9也明显减少。同时线粒体中的细胞色素C的释放也相应减少。结论高氧刺激可使肺上皮细胞Flot表达上调,其作用是保护细胞对抗高氧所造成的损伤,调控的分子机制与细胞的线粒体凋亡途径有关。
[Abstract]:Objective to investigate the regulatory molecular mechanism of Flotillin-2 Flot-2 in hyperoxia-induced apoptosis of pulmonary epithelial cells. Methods after the mice were exposed to hyperoxia for 24 h, the expression of Flot-2 was detected by immunohistochemistry. The expression of Flot-2 gene and protein in lung epithelial cell line after hyperoxia was further verified in vitro. The Flot-2 gene was integrated into the genome of pulmonary epithelial cells by plasmid transfection to make Flot-2 overexpression. Pulmonary epithelial cells were cultured in air and hyperoxia for 24 hours, and the total protein was extracted after collecting the cells. The effects of hyperoxia on apoptosis were investigated by Westen blot, including Caspase-3, Caspase-9, cytochrome C in mitochondria and cytoplasm. Results Flot-2 existed widely in bronchial epithelium and alveolar epithelial cells, and the expression of Flot-2 was significantly increased after hyperoxia stimulation. Pulmonary epithelial cells were stimulated by hyperoxia after transfection with high expression of Flot-2, and the cell death decreased, and the key apoptotic protein Caspase-3 and its active fragment, cleaved-Caspase-3, decreased significantly. The key apoptotic protein Caspase-9 and its active fragment cleaved-Caspase-9 in endogenous mitochondrial pathway were also significantly reduced. At the same time, the release of cytochrome C in mitochondria was also reduced. Conclusion hyperoxia stimulation can up-regulate the expression of Flot in lung epithelial cells, which protects the cells from the damage caused by hyperoxia. The molecular mechanism of hyperoxia regulation is related to the apoptosis pathway of mitochondria.
【作者单位】：广州市第一人民医院南沙医院病理科;广州市第一人民医院图书馆;广州市第一人民医院呼吸内科;

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