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首页> 外文期刊>Cellular Physiology and Biochemistry >Autophagy Activation by Rapamycin Before Hypoxia-Reoxygenation Reduces Endoplasmic Reticulum Stress in Alveolar Epithelial Cells
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Autophagy Activation by Rapamycin Before Hypoxia-Reoxygenation Reduces Endoplasmic Reticulum Stress in Alveolar Epithelial Cells

机译：低氧-再氧化前雷帕霉素的自噬激活减少了肺泡上皮细胞的内质网应激

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>Background: To determine potential effects of autophagy activation on hypoxia-reoxygenation (H/R) induced damage of a rat alveolar epithelial cell line. Methods: CCL149 cells were subjected to autophagy agonist (rapamycin, Rap), autophagy inhibitor (3-methyladenine, 3-MA) or PBS for 1 h before H/R treatment for 2 h, 4 h and 6 h. The optimal concentration of Rap (150 nM, 200 nM and 250 nM) or 3-MA (5 mM, 10 mM and 15 mM) was obtained from MTT assay. Autophagy was determined by fluorescence microscopy of eRFP-LC3 positive cells, transmission electron microscopy of autophagosome, western blot of LC3, AMPK, Beclin-1, HDAC6 and p62 proteins. Endoplasmatic reticulum stress was indicated by detecting expressions of BIP, XBP-1 and CHOP via western blot. Results: Rap at concentration of 250 nM before H/R increased the autophagy formation with more eRFP-LC3 positive cells and higher expressions of LC3-II, Beclin-1, HDAC6 and p62, but lower expressions of BIP, XBP-1 and CHOP in H/R treated CCL149. This effect seemed to be still obvious after H/R exposure for 6 h. The contrary results were obtained by treatment with 5 mM 3-MA. Conclusion: Rap might be a promising agent before mechanical ventilation or reperfusion to prevent re-damage in hypoxia related lung diseases.
机译：> 背景： 要确定自噬激活对缺氧-复氧（H / R）诱导的大鼠肺泡上皮细胞系损伤的潜在影响。 方法： 在H / R处理之前，将CCL149细胞置于自噬激动剂（雷帕霉素，Rap），自噬抑制剂（3-甲基腺嘌呤，3-MA）或PBS中1 h。持续2小时，4小时和6小时。从MTT分析中获得Rap（150 nM，200 nM和250 nM）或3-MA（5 mM，10 mM和15 mM）的最佳浓度。自噬是通过eRFP-LC3阳性细胞的荧光显微镜检查，自噬体的透射电子显微镜检查，LC3，AMPK，Beclin-1，HDAC6和p62蛋白的蛋白质印迹来确定的。通过Western印迹检测BIP，XBP-1和CHOP的表达来指示内质网应激。 结果： 在H / R之前，浓度为250 nM的Rap会增加自噬形成，其中eRFP-LC3阳性细胞更多，LC3-II，Beclin-1，HDAC6的表达更高和p62，但在经H / R处理的CCL149中BIP，XBP-1和CHOP的表达较低。 H / R暴露6 h后，这种效果似乎仍然很明显。通过用5 mM 3-MA处理获得相反的结果。 结论： 在机械通气或再灌注之前，Rap可能是一种有前途的药物，可防止缺氧相关的肺部疾病再次受损。

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