1. Resveratrol protects BV2 mouse microglial cells against LPS-induced inflammatory injury by altering the miR-146a-5p/TRAF6/NF-κB axis
- Author
-
Ying Lu, Yu-Ting Ge, An-Qi Zhong, and Guang-Fei Xu
- Subjects
Lipopolysaccharides ,0301 basic medicine ,Lipopolysaccharide ,Cell Survival ,Immunology ,Anti-Inflammatory Agents ,Cell Culture Techniques ,Resveratrol ,Toxicology ,Cell Line ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Western blot ,medicine ,Animals ,Immunology and Allergy ,MTT assay ,Viability assay ,TNF Receptor-Associated Factor 6 ,Pharmacology ,Dose-Response Relationship, Drug ,medicine.diagnostic_test ,NF-kappa B ,NF-κB ,General Medicine ,Transfection ,Molecular biology ,Up-Regulation ,MicroRNAs ,030104 developmental biology ,TNF receptor associated factor ,chemistry ,030220 oncology & carcinogenesis ,Cytokines ,Microglia ,Signal Transduction - Abstract
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.
- Published
- 2019
- Full Text
- View/download PDF