Deng Yuan, Wang Ganlan, Hou Dan, Zhang Lei, Pei Chaoying, Yang Guoshuai
Department of Neurology, Haikou Affiliated Hospital of Central South University Xiangya School of Medicine, Haikou, 570208, China.
In Vitro Cell Dev Biol Anim. 2025 Feb;61(2):178-188. doi: 10.1007/s11626-024-00986-0. Epub 2024 Dec 7.
MicroRNA-146a-5p (miR-146a-5p) actively participates in the process of cerebral ischemia-reperfusion (CI/R) injury. Dysregulation of the tumor necrosis factor receptor-associated factor 6 (TRAF6)/nuclear factor kappa-B (NF-κB) p65 axis is closely associated with inflammatory response. This study aimed to investigate the potential involvement of miR-146a-5p and TRAF6/NF-κB p65 in mediating CI/R progression in vitro. HT-22 cells were challenged with oxygen-glucose deprivation/reoxygenation (OGD/R) to simulate CI/R in vitro. HT-22 cells were transfected with miR-146a-5p mimics or TRAF6 overexpression constructs. The impact of miR-146a-5p on apoptosis, inflammation, and TRAF6/NF-κB p65 activation were investigated. OGD/R inhibited HT-22 cell viability, induced apoptosis, reduced miR-146a-5p levels and activated the TRAF6/NF-κB p65 pathway. MiR-146a-5p mimics reduced pro-inflammatory factor release, limited apoptosis-related protein expression, and inactivated the TRAF6/NF-κB p65 pathway in OGD/R-challenged HT-22 cells. Mechanistically, miR-146a-5p was verified to bind to TRAF6 3'UTR. TRAF6 overexpression reversed the beneficial effects of miR-146a-5p mimics on apoptosis, inflammation, and TRAF6/NF-κB p65 activation. This work revealed that miR-146a-5p targeted TRAF6 and suppressed the TRAF6/NF-κB p65 pathway, thereby reducing OGD/R-induced inflammation and apoptosis in HT-22 cells. These findings suggest the potential of the miR-146a-5p/TRAF6/NF-κB p65 axis in the treatment of CI/R.
微小RNA-146a-5p（miR-146a-5p）积极参与脑缺血再灌注（CI/R）损伤过程。肿瘤坏死因子受体相关因子6（TRAF6）/核因子κB（NF-κB）p65轴的失调与炎症反应密切相关。本研究旨在探讨miR-146a-5p和TRAF6/NF-κB p65在体外介导CI/R进展中的潜在作用。用氧糖剥夺/复氧（OGD/R）处理HT-22细胞以在体外模拟CI/R。用miR-146a-5p模拟物或TRAF6过表达构建体转染HT-22细胞。研究了miR-146a-5p对细胞凋亡、炎症和TRAF6/NF-κB p65激活的影响。OGD/R抑制HT-22细胞活力，诱导细胞凋亡，降低miR-146a-5p水平并激活TRAF6/NF-κB p65通路。miR-146a-5p模拟物减少促炎因子释放，限制凋亡相关蛋白表达，并使OGD/R处理的HT-22细胞中的TRAF6/NF-κB p65通路失活。机制上，验证了miR-146a-5p与TRAF6 3'非翻译区结合。TRAF6过表达逆转了miR-146a-5p模拟物对细胞凋亡、炎症和TRAF6/NF-κB p65激活的有益作用。这项工作表明，miR-146a-5p靶向TRAF6并抑制TRAF6/NF-κB p65通路，从而减少OGD/R诱导的HT-22细胞炎症和凋亡。这些发现提示miR-146a-5p/TRAF6/NF-κB p65轴在CI/R治疗中的潜力。
