Huc-MSCs-derived exosomes attenuate inflammatory pain by regulating microglia pyroptosis and autophagy via the miR-146a-5p/TRAF6 axis.
Huc-MSCs-derived exosomes attenuate inflammatory pain by regulating microglia pyroptosis and autophagy via the miR-146a-5p/TRAF6 axis.
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Huc-MSC 衍生的外泌体通过 miR-146a-5p/TRAF6 轴调节小胶质细胞焦亡和自噬，从而减轻炎症疼痛。
DOI：
10.1186/s12951-022-01522-6
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发表时间：
2022-07-14
影响因子：
10.2
通讯作者：
Yuan, Hongbin
中科院分区：
文献类型：
--
作者：
Hua, Tong;Yang, Mei;Song, Honghao;Kong, Erliang;Deng, Mengqiu;Li, Yongchang;Li, Jian;Liu, Zhixiao;Fu, Hailong;Wang, Yue;Yuan, Hongbin
慢性炎性疼痛显著降低生活质量，缺乏有效的干预措施。近年来，人脐带间充质干细胞（human umbilical cord mesenchymal stem cells，huc-MSCs）衍生的外泌体作为一种有前途的无细胞治疗策略已被用于缓解神经病理性疼痛和其他炎性疾病。然而，huc-MSC衍生的外泌体在完全弗氏佐剂（CFA）诱导的炎性疼痛中的治疗价值仍有待证实。在这项研究中，我们研究了huc-MSCs来源的外泌体在慢性炎症疼痛模型中的治疗效果和相关机制。采用C57 BL/6 J雄性小鼠建立CFA诱导的炎性疼痛模型，鞘内注射huc-MSCs来源的exosomes，连续4天。用脂多糖（LPS）加三磷酸腺苷（ATP）刺激BV 2小胶质细胞，以研究huc-MSC衍生的外泌体对焦亡和自噬的影响。使用生物信息学分析和拯救实验来证明miR-146 a-5 p/TRAF 6在调节焦亡和自噬中的作用。采用Western blotting、RT-qPCR、small interfering RNA和Yo-Pro-1染料染色等方法研究其作用机制。Huc-MSC衍生的外泌体减轻CFA诱导的炎性疼痛中的机械异常性疼痛和热痛觉过敏。此外，huc-MSCs衍生的外泌体通过增加自噬相关蛋白（LC 3-II和beclin 1）的表达和抑制脊髓背角中NLRP 3炎性体的激活来减轻神经炎症。在体外，在用huc-MSC衍生的外泌体预处理的BV 2细胞中，NLRP 3炎性体组分（NLRP 3、半胱天冬酶1-p20、ASC）和gasdermin D（GSDMD-F、GSDMD-N）被抑制。Western blot和Yo-Pro-1染料染色结果表明，自噬抑制剂3-MA可减弱huc-MSCs来源的exosomes对BV 2细胞凋亡的保护作用。重要的是，用miR-146 a-5 p模拟物转染的huc-MSC衍生的外泌体促进自噬并抑制BV 2细胞焦亡。TRAF 6作为miR-146 a-5 p的靶基因，通过小干扰RNA（small interfering RNA，siRNA）被敲低，从而增加焦亡并抑制自噬。Huc-MSC衍生的外泌体通过miR-146 a-5 p/TRAF 6减轻炎性疼痛，这增加了自噬水平并抑制了焦亡。在线版本包含补充材料，可通过10.1186/s12951-022-01522-6获得。
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. 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. 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. Huc-MSCs-derived exosomes attenuated inflammatory pain via miR-146a-5p/TRAF6, which increased the level of autophagy and inhibited pyroptosis. The online version contains supplementary material available at 10.1186/s12951-022-01522-6.
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影响因子：
4.6
作者：
Asami T;Ishii M;Fujii H;Namkoong H;Tasaka S;Matsushita K;Ishii K;Yagi K;Fujiwara H;Funatsu Y;Hasegawa N;Betsuyaku T
通讯作者：
Betsuyaku T
M1巨噬细胞衍生的细胞外囊泡传递miR-146a-5p和miR-146b-5p，通过靶向TRAF6抑制复发性流产中的滋养层细胞迁移和侵袭
影响因子：
12.4
作者：
Ding J;Zhang Y;Cai X;Zhang Y;Yan S;Wang J;Zhang S;Yin T;Yang C;Yang J
通讯作者：
Yang J
影响因子：
3.3
作者：
Joerger-Messerli MS;Oppliger B;Spinelli M;Thomi G;di Salvo I;Schneider P;Schoeberlein A
通讯作者：
Schoeberlein A
神经炎症，骨髓干细胞和慢性疼痛。
影响因子：
7.3
作者：
Huh Y;Ji RR;Chen G
通讯作者：
Chen G
神经炎症驱动的慢性疼痛的新兴目标。
影响因子：
120.1
作者：
Ji, Ru-Rong;Xu, Zhen-Zhong;Gao, Yong-Jing
通讯作者：
Gao, Yong-Jing