MiR-367 alleviates inflammatory injury of microglia by promoting M2 polarization via targeting CEBPA
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MiR-367 alleviates inflammatory injury of microglia by promoting M2 polarization via targeting CEBPA Hui Pei 1 & Qian Peng 2 & Shewei Guo 3 & Yulei Gu 1 & Tongwen Sun 4 & Dong Xu 1 & Yumin Jiang 1 & Jiafeng Xie 1 & Luanluan Zhang 1 & Zhiqiang Zhu 1 Received: 14 February 2020 / Accepted: 1 October 2020 / Editor: Tetsuji Okamoto # The Author(s) 2020
Abstract MiR-367 was reported to regulate inflammatory response of microglia. CCAAT/enhancer-binding protein α (C/EBPA) could mediate microglia polarization. In this study, we explored the possible roles of miR-367 and CEBPA in intracerebral hemorrhage (ICH). ICH and normal specimens were obtained from the tissue adjacent to and distant from hematoma of ICH patients, respectively. Microglia were isolated and identified by immunofluorescence. The isolated microglia were treated with erythrocyte lysate and randomly divided into 8 groups using different transfection reagents. The transfection efficiency of miR-367 was determined by qRT-PCR. The expressions of M1 and M2 microglia markers were detected by Western blotting. The relationship between CEBPA and miR-367 was confirmed by dual luciferase reporter system. Flow cytometry was performed to determine the level of apoptosis in the cells transfected with miR-367 and CEBPA in erythrocyte lysate–treated microglia. We found that miR367 expression level was downregulated in ICH specimens. Erythrocyte lysate–treated microglia was successfully established using erythrocyte lysate, as decreased miR-367 expression was observed. Overexpression of miR-367 could significantly decrease the expressions of MHC-ІІ, IL-1β, and Bax, reduced apoptosis rate, and increased the expressions of CD206, Bal-2, and Arg-1 in erythrocyte lysate–treated microglia. CEBPA was proved to be a direct target for miR-367, which could inhibit microglia M2 polarization and increase apoptosis rate. However, in the presence of both CEBPA and miR-367 mimic, the protein and mRNA expressions of CEBPA were decreased, leading to promoted microglia M2 polarization and a decreased apoptosis rate. MiR-367 regulates microglia polarization by targeting CEBPA and is expected to alleviate ICH-induced inflammatory injury. Keywords miR-367 . M2 polarization . Microglia . CEBPA . Intracerebral hemorrhage
Introduction Hemorrhagic stroke (also known as intracerebral hemorrhage (ICH)) is the most acute and serious cerebrovascular disease,
Hui Pei and Qian Peng contributed equally to this work. * Zhiqiang Zhu [email protected] 1
Department of Emergency Intensive Care Unit, the First Affiliated Hospital of Zhengzhou University, No.1, East Jianshe Road, Erqi District, Zhengzhou 450052, Henan Province, China
2
Department of Plastic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China
3
Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China
4
Department of General Intensive Care Unit, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China
as it has a quick onset and high mortality a
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