Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou, P.R. China.
Department of Anesthesiology, Guizhou Province People's Hospital, Guiyang, P.R. China.
Aging (Albany NY). 2022 Aug 16;14(16):6507-6519. doi: 10.18632/aging.204137.
Some microRNAs (miRNAs) play important roles in lung ischemia-reperfusion injury (LIRI) injury. Here, this study aimed to examine whether miR-141 was related to lung ischemia-reperfusion injury (IRI) via regulating autophagy and the epidermal growth factor receptor (EGFR), and to explore the underlying signal transduction pathways. To this end, we constructed the LIRI cell model and mouse models, separately. According to RT-qPCR and Western blotting (WB) analysis results, miR-141 up-regulation together with β-catenin and EGFR down-regulation within mouse pulmonary microvascular endothelial cells (PMVECs) or lung tissues was related to lung IRI. Besides, we conducted dual-luciferase reporter assay, which suggested the binding of EGFR to miR-141. In addition, we carried out TUNEL staining, HE staining, and flow cytometric analysis to assess the apoptosis of PMVECs and the injury to mouse lung tissues. Furthermore, we performed light-chain immunofluorescence assay to examine autophagosomes within PMVECs. According to our results, miR-141 suppressed β-catenin level through reducing EGFR level. Besides, the miR-141/EGFR/β-catenin axis enhanced autophagy to aggravate LIRI. To sum up, miR-141 suppresses EGFR expression to inhibit β-catenin level, which subsequently aggravates autophagy and complicates LIRI. The above results offer the candidate therapeutic target for the treatment of lung IRI.
一些 microRNAs（miRNAs）在肺缺血再灌注损伤（LIRI）损伤中发挥重要作用。本研究旨在探讨 miR-141 是否通过调节自噬和表皮生长因子受体（EGFR）与肺缺血再灌注损伤（IRI）有关，并探讨其潜在的信号转导途径。为此，我们分别构建了 LIRI 细胞模型和小鼠模型。根据 RT-qPCR 和 Western blot（WB）分析结果，在小鼠肺微血管内皮细胞（PMVECs）或肺组织中，miR-141 的上调以及β-catenin 和 EGFR 的下调与肺 IRI 有关。此外，我们进行了双荧光素酶报告基因检测，表明 EGFR 与 miR-141 结合。另外，我们进行了 TUNEL 染色、HE 染色和流式细胞术分析，以评估 PMVECs 的凋亡和小鼠肺组织的损伤。此外，我们进行了轻链免疫荧光检测，以检测 PMVECs 中的自噬体。根据我们的结果，miR-141 通过降低 EGFR 水平来抑制β-catenin 水平。此外，miR-141/EGFR/β-catenin 轴增强自噬作用，加重 LIRI。总之，miR-141 抑制 EGFR 表达以抑制β-catenin 水平，进而加重自噬并使 LIRI 复杂化。上述结果为治疗肺 IRI 提供了候选治疗靶点。
