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 miR-367 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.

据报道，MiR-367 可调节小胶质细胞的炎症反应。CCAAT/增强子结合蛋白α（C/EBPA）可以介导小胶质细胞极化。在这项研究中，我们探讨了 miR-367 和 CEBPA 在脑出血 (ICH) 中的可能作用。ICH和正常标本分别取自ICH患者血肿附近和远处的组织。通过免疫荧光分离和鉴定小胶质细胞。分离出的小胶质细胞用红细胞裂解液处理，并使用不同的转染试剂随机分为 8 组。通过qRT-PCR测定miR-367的转染效率。Western印迹检测M1和M2小胶质细胞标志物的表达。CEBPA和miR-367之间的关系由双荧光素酶报告系统证实。进行流式细胞术以确定在红细胞裂解物处理的小胶质细胞中用 miR-367 和 CEBPA 转染的细胞的凋亡水平。我们发现 miR-367 表达水平在 ICH 标本中下调。使用红细胞裂解物成功建立了红细胞裂解物处理的小胶质细胞，因为观察到 miR-367 表达降低。miR-367的过表达可显着降低红细胞裂解液处理的小胶质细胞中MHC-ІІ、IL-1β和Bax的表达，降低细胞凋亡率，并增加CD206、Bal-2和Arg-1的表达。CEBPA被证明是miR-367的直接靶点，可以抑制小胶质细胞M2极化并增加细胞凋亡率。然而，在 CEBPA 和 miR-367 模拟物同时存在的情况下，CEBPA 的蛋白质和 mRNA 表达降低，导致促进小胶质细胞 M2 极化和降低细胞凋亡率。MiR-367 通过靶向 CEBPA 调节小胶质细胞极化，有望减轻 ICH 诱导的炎症损伤。