Objective: To investigate the role of miR-146a-5p in the effects of resveratrol (RSV) on inflammatory response in BV2 mouse microglial cells. Materials and methods: BV2 cells were pretreated by RSV and stimulated with lipopolysaccharide (LPS). Cell Viability was checked using a MTT assay. Real-Time PCR was performed to detect the levels of pro-inflammatory cytokines (tumor necrosisfactor-α－TNF-α, interleukin-1β－IL-1β and interleukin-6 － IL-6) and miR-146a-5p expression. Western blot was used to analyze the protein expression of TNF receptor associated factor 6 (TRAF6) and phospho-nuclear factor kappa B (pNF-κB). Gain-of-function and loss-of-function analysis of miR-146a-5p was performed using transfection of miR-146a-5p mimic and miR-146a-5p inhibitor, respectively. Results: Pretreatment with RSV significantly and dose dependently inhibited LPS-induced production of TNF-α, IL-1β and IL-6 in BV2 cells. MiR-146a-5p was significantly upregulated after LPS treatment, and further increased in RSV and LPS-co-treated cells. MiR-146a-5p overexpression via miR-146a-5p mimic transfection downregulated the mRNA level of TNF-α, IL-1β and IL-6, as well as abrogated the protein expression of TRAF6 and pNF-κB in BV2 cells exposed to LPS. More importantly, the reducion of TNF-α, IL-1β and IL-6 level by RSV were reversed by miR-146a-5p silence via miR-146a-5p inhibitor transfection. Furthermore, silencing miR-146a-5p attenuated the inhibitory effect of RSV on the TRAF6/NF-κB pathway which was activated after induction with LPS. Conclusions: RSV can suppress LPS-induced inflammatory injury via modulating the miR-146a-5p/TRAF6/NF-κB axis in BV2 mouse microglial cells.

中文翻译：

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白藜芦醇通过改变miR-146a-5p / TRAF6 /NF-κB轴，保护BV2小鼠小胶质细胞免受LPS诱导的炎症损伤。

目的：探讨miR-146a-5p在白藜芦醇（RSV）对BV2小鼠小胶质细胞炎症反应中的作用。材料和方法：BV2细胞经RSV预处理，并用脂多糖（LPS）刺激。使用MTT测定法检查细胞活力。实时荧光定量PCR检测促炎细胞因子（肿瘤坏死因子-α-TNF-α，白介素-1β-IL-1β和白介素-6-IL-6）和miR-146a-5p表达。用蛋白质印迹法分析了TNF受体相关因子6（TRAF6）和磷酸核因子κB（pNF-κB）的蛋白表达。使用miR-146a-5p模拟物和miR-146a-5p抑制剂转染分别进行miR-146a-5p的功能获得和功能丧失分析。结果：用RSV预处理可以显着且剂量依赖性地抑制LPS诱导的BV2细胞中TNF-α，IL-1β和IL-6的产生。LPS处理后，MiR-146a-5p显着上调，并在RSV和LPS共处理的细胞中进一步增加。通过miR-146a-5p模拟转染的MiR-146a-5p过表达下调了LPS暴露的BV2细胞中TNF-α，IL-1β和IL-6的mRNA水平，并废除了TRAF6和pNF-κB的蛋白表达。 。更重要的是，通过miR-146a-5p抑制剂转染的miR-146a-5p沉默可以逆转RSV对TNF-α，IL-1β和IL-6水平的降低。此外，沉默miR-146a-5p减弱了RSV对TRAP6 /NF-κB通路的抑制作用，该通路在LPS诱导后被激活。结论：