Dorsal root ganglion-derived exosomes deteriorate neuropathic pain by activating microglia via the microRNA-16-5p/HECTD1/HSP90 axis.

IF 4.3 2区生物学 Q1 BIOLOGY Biological Research Pub Date : 2024-05-15 DOI:10.1186/s40659-024-00513-1

Yinghao Xing, Pei Li, Yuanyuan Jia, Kexin Zhang, Ming Liu, Jingjing Jiang

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Abstract

Background: The activated microglia have been reported as pillar factors in neuropathic pain (NP) pathology, but the molecules driving pain-inducible microglial activation require further exploration. In this study, we investigated the effect of dorsal root ganglion (DRG)-derived exosomes (Exo) on microglial activation and the related mechanism.

Methods: A mouse model of NP was generated by spinal nerve ligation (SNL), and DRG-derived Exo were extracted. The effects of DRG-Exo on NP and microglial activation in SNL mice were evaluated using behavioral tests, HE staining, immunofluorescence, and western blot. Next, the differentially enriched microRNAs (miRNAs) in DRG-Exo-treated microglia were analyzed using microarrays. RT-qPCR, RNA pull-down, dual-luciferase reporter assay, and immunofluorescence were conducted to verify the binding relation between miR-16-5p and HECTD1. Finally, the effects of ubiquitination modification of HSP90 by HECTD1 on NP progression and microglial activation were investigated by Co-IP, western blot, immunofluorescence assays, and rescue experiments.

Results: DRG-Exo aggravated NP resulting from SNL in mice, promoted the activation of microglia in DRG, and increased neuroinflammation. miR-16-5p knockdown in DRG-Exo alleviated the stimulating effects of DRG-Exo on NP and microglial activation. DRG-Exo regulated the ubiquitination of HSP90 through the interaction between miR-16-5p and HECTD1. Ubiquitination alteration of HSP90 was involved in microglial activation during NP.

Conclusions: miR-16-5p shuttled by DRG-Exo regulated the ubiquitination of HSP90 by interacting with HECTD1, thereby contributing to the microglial activation in NP.

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背根神经节衍生的外泌体通过microRNA-16-5p/HECTD1/HSP90轴激活小胶质细胞，从而恶化神经病理性疼痛。

背景：据报道，活化的小胶质细胞是神经病理性疼痛（NP）病理学的支柱因素，但驱动疼痛诱导的小胶质细胞活化的分子还需要进一步探索。本研究探讨了背根神经节（DRG）衍生的外泌体（Exo）对小胶质细胞活化的影响及其相关机制：方法：通过脊神经结扎术（SNL）建立NP小鼠模型，并提取DRG衍生的外泌体。通过行为测试、HE染色、免疫荧光和Western blot等方法评估了DRG-Exo对SNL小鼠NP和小胶质细胞活化的影响。接着，使用芯片分析了 DRG-Exo 处理过的小胶质细胞中不同程度富集的微 RNA（miRNA）。通过 RT-qPCR、RNA pull-down、双荧光素酶报告实验和免疫荧光来验证 miR-16-5p 与 HECTD1 的结合关系。最后，通过Co-IP、Western印迹、免疫荧光检测和挽救实验研究了HECTD1对HSP90泛素化修饰对NP进展和小胶质细胞活化的影响：结果：DRG-Exo加重了小鼠SNL导致的NP，促进了DRG中小胶质细胞的活化，加重了神经炎症。DRG-Exo 通过 miR-16-5p 和 HECTD1 之间的相互作用调节 HSP90 的泛素化。结论：DRG-Exo穿梭的miR-16-5p通过与HECTD1相互作用调节了HSP90的泛素化，从而促进了NP中的小胶质细胞活化。

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来源期刊

Biological Research 生物-生物学

CiteScore

10.10

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Biological Research is an open access, peer-reviewed journal that encompasses diverse fields of experimental biology, such as biochemistry, bioinformatics, biotechnology, cell biology, cancer, chemical biology, developmental biology, evolutionary biology, genetics, genomics, immunology, marine biology, microbiology, molecular biology, neuroscience, plant biology, physiology, stem cell research, structural biology and systems biology.