Teng Jiajia, Yang Feng, Li Xiaoling
Department of Nephropathy, Yantai Hospital of Traditional Chinese Medicine, Yantai, Shandong 264001, P.R. China.
Department of Rheumatology, Yantai Hospital of Traditional Chinese Medicine, Yantai, Shandong 264001, P.R. China.
Exp Ther Med. 2022 Nov 22;25(1):21. doi: 10.3892/etm.2022.11720. eCollection 2023 Jan.
Dysregulated microRNA (miRNA or miR) expression is an important cause of immune homeostasis disorder in patients with systemic lupus erythematosus and lupus nephritis (LN). The present study evaluated the possibility of using miR-146a-5p as a therapeutic target for treating LN. The effects of miR-146a-5p on lupus syndrome in MRL/lpr mice were evaluated. MRL/lpr mice were injected with miR-146a-5p agomir (M146AG) or agomir negative control (NC). Renal function index, pathology and protein expression levels of inflammatory factors in MRL/lpr mice were evaluated after M146AG or agomir NC treatment. Reverse transcription-quantitative PCR, western blotting and immunofluorescence were used to assess the effect of M146AG on mRNA and protein expression levels of (tumor necrosis factor receptor-associated factor 6) TRAF6/NF-κB axis components. A luciferase dual reporter system was used to assess the mechanism of regulation of TRAF6/NF-κB axis expression. Finally, flow cytometry was used to assess the regulatory effect of M146AG on regulatory T cell (Treg)/T helper 17 (Th17) balance. The findings demonstrated that M146AG ameliorated renal lesions and the inflammatory response in MRL/lpr mice. TRAF6 was demonstrated to be targeted and significantly negatively regulated by miR-146a-5p. M146AG intervention significantly increased expression of miR-146a-5p and significantly downregulated the mRNA and protein expression levels of TRAF6 and NF-κB in CD4 T cells of MRL/lpr mice. Furthermore, M146AG intervention alleviated Treg/Th17 imbalance in MRL/lpr mice peripheral blood. The present findings demonstrated that M146AG improved Treg/Th17 imbalance and alleviated renal lesions in MRL/lpr mice by targeting the TRAF6/NF-κB axis. This may provide a new theoretical basis for the clinical diagnosis and treatment of LN.
微小RNA（miRNA或miR）表达失调是系统性红斑狼疮和狼疮性肾炎（LN）患者免疫稳态紊乱的重要原因。本研究评估了将miR-146a-5p用作治疗LN的治疗靶点的可能性。评估了miR-146a-5p对MRL/lpr小鼠狼疮综合征的影响。给MRL/lpr小鼠注射miR-146a-5p激动剂（M146AG）或激动剂阴性对照（NC）。在M146AG或激动剂NC处理后，评估MRL/lpr小鼠的肾功能指标、病理学以及炎症因子的蛋白表达水平。采用逆转录定量PCR、蛋白质印迹法和免疫荧光法评估M146AG对（肿瘤坏死因子受体相关因子6）TRAF6/NF-κB轴成分的mRNA和蛋白表达水平的影响。使用荧光素酶双报告系统评估TRAF6/NF-κB轴表达的调控机制。最后，采用流式细胞术评估M146AG对调节性T细胞（Treg）/辅助性T细胞17（Th17）平衡的调节作用。研究结果表明，M146AG改善了MRL/lpr小鼠的肾脏病变和炎症反应。结果表明TRAF6是miR-146a-5p的靶标且受到其显著负调控。M146AG干预显著增加了miR-146a-5p的表达，并显著下调了MRL/lpr小鼠CD4 T细胞中TRAF6和NF-κB的mRNA和蛋白表达水平。此外，M146AG干预减轻了MRL/lpr小鼠外周血中的Treg/Th17失衡。本研究结果表明，M146AG通过靶向TRAF6/NF-κB轴改善了MRL/lpr小鼠的Treg/Th17失衡并减轻了肾脏病变。这可能为LN的临床诊断和治疗提供新的理论依据。
