PMID- 35836229 OWN - NLM STAT- MEDLINE DCOM- 20220718 LR - 20220718 IS - 1477-3155 (Electronic) IS - 1477-3155 (Linking) VI - 20 IP - 1 DP - 2022 Jul 14 TI - Huc-MSCs-derived exosomes attenuate inflammatory pain by regulating microglia pyroptosis and autophagy via the miR-146a-5p/TRAF6 axis. PG - 324 LID - 10.1186/s12951-022-01522-6 [doi] LID - 324 AB - BACKGROUND: Chronic inflammatory pain significantly reduces the quality of life and lacks effective interventions. In recent years, human umbilical cord mesenchymal stem cells (huc-MSCs)-derived exosomes have been used to relieve neuropathic pain and other inflammatory diseases as a promising cell-free therapeutic strategy. However, the therapeutic value of huc-MSCs-derived exosomes in complete Freund's adjuvant (CFA)-induced inflammatory pain remains to be confirmed. In this study, we investigated the therapeutic effect and related mechanisms of huc-MSCs-derived exosomes in a chronic inflammatory pain model. METHODS: C57BL/6J male mice were used to establish a CFA-induced inflammatory pain model, and huc-MSCs-derived exosomes were intrathecally injected for 4 consecutive days. BV2 microglia cells were stimulated with lipopolysaccharide (LPS) plus adenosine triphosphate (ATP) to investigate the effect of huc-MSCs-derived exosomes on pyroptosis and autophagy. Bioinformatic analysis and rescue experiments were used to demonstrate the role of miR-146a-5p/ TRAF6 in regulating pyroptosis and autophagy. Western blotting, RT-qPCR, small interfering RNA and Yo-Pro-1 dye staining were performed to investigate the related mechanisms. RESULTS: Huc-MSCs-derived exosomes alleviated mechanical allodynia and thermal hyperalgesia in CFA-induced inflammatory pain. Furthermore, huc-MSCs-derived exosomes attenuated neuroinflammation by increasing the expression of autophagy-related proteins (LC3-II and beclin1) and inhibiting the activation of NLRP3 inflammasomes in the spinal cord dorsal horn. In vitro, NLRP3 inflammasome components (NLRP3, caspase1-p20, ASC) and gasdermin D (GSDMD-F, GSDMD-N) were inhibited in BV2 cells pretreated with huc-MSCs-derived exosomes. Western blot and Yo-Pro-1 dye staining demonstrated that 3-MA, an autophagy inhibitor, weakened the protective effect of huc-MSCs-derived exosomes on BV2 cell pyroptosis. Importantly, huc-MSCs-derived exosomes transfected with miR-146a-5p mimic promoted autophagy and inhibited BV2 cell pyroptosis. TRAF6, as a target gene of miR-146a-5p, was knocked down via small-interfering RNA, which increased pyroptosis and inhibited autophagy. CONCLUSION: Huc-MSCs-derived exosomes attenuated inflammatory pain via miR-146a-5p/TRAF6, which increased the level of autophagy and inhibited pyroptosis. CI - (c) 2022. The Author(s). FAU - Hua, Tong AU - Hua T AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Yang, Mei AU - Yang M AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Song, Honghao AU - Song H AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Kong, Erliang AU - Kong E AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Deng, Mengqiu AU - Deng M AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Li, Yongchang AU - Li Y AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Li, Jian AU - Li J AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. FAU - Liu, Zhixiao AU - Liu Z AD - Research Center of Developmental Biology, Department of Histology and Embryology, College of Basic Medicine, Naval Medical University, Shanghai, 200433, China. FAU - Fu, Hailong AU - Fu H AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. fuhailong1979@163.com. FAU - Wang, Yue AU - Wang Y AD - Stem Cell and Regeneration Medicine Institute, Research Center of Translational Medicine, Naval Medical University, Shanghai, 200433, China. wangyuesmmu@163.com. FAU - Yuan, Hongbin AU - Yuan H AD - Department of Anesthesiology, Changzheng Hospital, Naval Medical University, Shanghai, 200003, China. jfjczyy@163.com. LA - eng GR - 81971046/National Natural Science Foundation of China/ GR - 82171220/National Natural Science Foundation of China/ PT - Journal Article DEP - 20220714 PL - England TA - J Nanobiotechnology JT - Journal of nanobiotechnology JID - 101152208 RN - 0 (MicroRNAs) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (TNF Receptor-Associated Factor 6) SB - IM MH - Animals MH - Autophagy MH - *Exosomes/metabolism MH - Humans MH - Male MH - Mice MH - Mice, Inbred C57BL MH - *MicroRNAs/genetics/metabolism MH - Microglia/metabolism MH - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism MH - Pain MH - Pyroptosis MH - Quality of Life MH - TNF Receptor-Associated Factor 6/genetics/metabolism PMC - PMC9281091 OTO - NOTNLM OT - Autophagy OT - Huc-MSCs-derived exosomes OT - Inflammatory pain OT - Microglia OT - Pyroptosis COIS- The authors declare that they have no competing interests. EDAT- 2022/07/15 06:00 MHDA- 2022/07/19 06:00 PMCR- 2022/07/14 CRDT- 2022/07/14 23:44 PHST- 2022/04/20 00:00 [received] PHST- 2022/06/21 00:00 [accepted] PHST- 2022/07/14 23:44 [entrez] PHST- 2022/07/15 06:00 [pubmed] PHST- 2022/07/19 06:00 [medline] PHST- 2022/07/14 00:00 [pmc-release] AID - 10.1186/s12951-022-01522-6 [pii] AID - 1522 [pii] AID - 10.1186/s12951-022-01522-6 [doi] PST - epublish SO - J Nanobiotechnology. 2022 Jul 14;20(1):324. doi: 10.1186/s12951-022-01522-6.