细胞自噬在大鼠Ⅱ型肺泡上皮细胞低氧/复氧损伤中的作用

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (10): 1440-1445.

细胞自噬在大鼠Ⅱ型肺泡上皮细胞低氧/复氧损伤中的作用

刘秀洁¹, 王新雨¹, 宋正阳¹, 楼国强¹, 郝卯林¹, 钱巍², 王万铁^1*

1.温州医科大学缺血/再灌注损伤研究所,浙江温州 325035;
2.杭州临安区人民医院全科医疗科, 浙江杭州 311300

收稿日期:2021-04-30 修回日期:2021-07-20 发布日期:2021-09-29
通讯作者: *wwt@wmu.edu.cn
基金资助:
温州市基础性科研项目(Y20180093);杭州市社会发展科研入库立项补助项目(201811)

Role of autophagy in hypoxia-reoxygenation-induced injury of rat alveolar epithelial cell type Ⅱ

LIU Xiu-jie¹, WANG Xin-yun¹, SONG Zheng-yang¹, LOU Guo-qiang¹, HAO Mao-lin¹, QIAN Wei², WANG Wan-tie^1*

1. Institute of Ischemia/Reperfusion Injury, Wenzhou Medical University, Wenzhou 325035;
2. Department of General Medicine, Lin'an District People's Hospital, Hangzhou 311300, China

Received:2021-04-30 Revised:2021-07-20 Published:2021-09-29
Contact: *wwt@wmu.edu.cn

摘要/Abstract

摘要： 目的探讨细胞自噬在大鼠Ⅱ型肺泡上皮细胞(AEC Ⅱ)低氧/复氧(H/R)中的作用。方法将大鼠AEC Ⅱ分为对照组(C)、低氧/复氧组(H/R)、DMSO溶剂组(HD)、自噬激动剂组(Rap)和自噬抑制剂组(3-MA)。CCK-8法检测细胞活力;反转录RT-PCR检测自噬相关蛋白mRNA表达水平;Western blot检测自噬相关蛋白水平;透射电镜检测细胞超微结构。结果与对照组相比,H/R组细胞活力均有下降,自噬相关蛋白及mRNA表达水平均上调(P＜0.05),细胞超微结构损伤较严重;与H/R组相比,3-MA组的细胞活力较高,自噬相关蛋白及mRNA表达水平均下调(P＜0.05)。结论低氧/复氧可激活细胞自噬引发大鼠肺泡Ⅱ型上皮细胞损伤。

关键词: 低氧/复氧损伤, 细胞自噬, Ⅱ型肺泡上皮细胞

Abstract: Objective To investigate the role of autophagy in the hypoxia-reoxygenation (H/R) model of rat alveolar epithelial cell type Ⅱ (AEC Ⅱ). Methods Cells were randomly divided into control group (C),hypoxia-reoxygenation group (H/R),solvent group (HD),autophagy agonist group(Rap) and autophagy inhibitor group (3-MA).Pretreated cells with 3-mA 1h before hypoxia. The cell viability was determined with CCK-8, and used RT-PCR, Western blot to detect the expression of autophagy related mRNA transmission and its proteins, and ultrastructure of cell was examined by electron microscopy. Results Compared to the C group, the cell viability decreased, the expression of autophagy related proteins and gene transcription increased, and the cell ultra-structure was significantly damaged (P＜0.05). The cell viability in 3-MA group was higher than that of H/R group, the autophagy related proteins and their gene transcription were all decreased (P＜0.05), the cell structure was damaged slightly,and autophagy body was not found. Conclusions Hypoxia-reoxygenation can activate autophagy and induce alveolar epithelial cell type Ⅱ damage.

Key words: hypoxia-reoxygenation injury, autophagy, alveolar epithelial cell type Ⅱ

中图分类号:

R363

刘秀洁, 王新雨, 宋正阳, 楼国强, 郝卯林, 钱巍, 王万铁. 细胞自噬在大鼠Ⅱ型肺泡上皮细胞低氧/复氧损伤中的作用[J]. 基础医学与临床, 2021, 41(10): 1440-1445.

LIU Xiu-jie, WANG Xin-yun, SONG Zheng-yang, LOU Guo-qiang, HAO Mao-lin, QIAN Wei, WANG Wan-tie. Role of autophagy in hypoxia-reoxygenation-induced injury of rat alveolar epithelial cell type Ⅱ[J]. Basic & Clinical Medicine, 2021, 41(10): 1440-1445.

参考文献

[1]Chen F, Date H. Update on ischemia-reperfusion injury in lung transplantation[J]. Curr Opin Organ Transplant, 2015, 20: 515-520.
[2]崔淑侠. 黄芪注射液对肺血栓栓塞症及其溶栓后再灌注损伤的影响 [D]. 武汉大学, 2018:70-79.
[3]李百玲, 柴家科, 段红杰,等. 肺泡Ⅱ型上皮细胞在急性肺损伤/急性呼吸窘迫综合征中的研究进展[J]. 中华损伤与修复杂志(电子版), 2014, 9: 323-325.
[4]陈鑫. 缺氧后处理影响肺泡Ⅱ型上皮细胞死亡受体途径凋亡机制的研究 [D].武汉:遵义医学院, 2012:1-41.
[5]刘浩, 刘修恒, 王磊, 等. 自噬在肾缺血再灌注损伤中的研究进展[J]. 职业与健康, 2019, 35: 700-704.
[6]李子一, 佟立权. 自噬在肝脏缺血再灌注损伤中的作用及研究进展[J]. 世界华人消化杂志, 2018, 26: 755-759.
[7]王彤, 李敬敬, 薛桥臻, 等. 细胞自噬在脑缺血再灌注损伤中作用机制的研究进展[J]. 世界最新医学信息文摘, 2019, 19: 71-72+74.
[8]田国卫, 夏豪. 自噬在心肌缺血再灌注损伤中的研究进展[J]. 疑难病杂志, 2018, 17: 1179-1184.
[9]李少莹. 自噬在LPS诱导的肺泡上皮细胞死亡中的作用及机制研究 [D].重庆:第三军医大学, 2015:25-39.
[10]Kim J, Kundu M, Viollet B, et al. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1[J]. Nat Cell Biol, 2011, 13: 132-141.
[11]Kang EB, Cho JY. Effect of treadmill exercise on PI3K/AKT/mTOR, autophagy, and Tau hyperphosphorylation in the cerebral cortex of NSE/htau23 transgenic mice[J]. J Exerc Nutrition Biochem, 2015, 19: 199-209.
[12]Luo T, Zhang H, Yu Q, et al. ERK1/2 MAPK promotes autophagy to suppress ER stress-mediated apoptosis induced by cadmium in rat proximal tubular cells[J]. Toxicol Vitro, 2018,52: 60-69.
[13]Benjamin D, Colombi M, Moroni C, et al. Rapamycin passes the torch: a new generation of mTOR inhibitors[J]. Nat Rev Drug Discov, 2011,10:868-880.
[14]Tanida I, Waguri S. Measurement of autophagy in cells and tissues[J]. Methods Mol Biol, 2010, 648: 193-214.
[15]Vittonatto E, Boschi S, Mongini T, et al. Differential diagnosis of vacuolar muscle biopsies: use of p62, LC3 and LAMP2 immunohistochemistry[J]. Acta Myol, 2017, 36:191-198.