Bone marrow mesenchymal stem cell-derived exosomes shuttling miR-150-5p alleviates mechanical allodynia in rats by targeting NOTCH2 in microglia

Abstract

Background This study probes into the function and mechanism of bone marrow mesenchymal stem cell (BMSC)-derived exosomes loaded with miR-150-5p in mechanical allodynia. Methods BMSCs were infected with miR-150-5p inhibition lentiviruses to obtain exosomes with low miR-150-5p expression. A L5 spinal nerve ligation (SNL) model was established in rats where exosomes, NOTCH2 overexpression/inhibition plasmids, or microglial cells were intrathecally administered. Hind paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) of rats were measured. TUNEL staining was used to measure the apoptotic rate in rat spinal dorsal horn (SDH), ELISA to evaluate pro-inflammatory factor levels, and RT-qPCR, western blotting, and immunohistochemistry to detect miR-150-5p and NOTCH2 expression. Immunofluorescence was used for localizing exosomes and NOTCH2 and detecting the expression of OX42, a maker for microglia. Dual luciferase reporter and RNA pull down assays were performed to validate the putative binding between miR-150-5p and NOTCH2. Results NOTCH2 expressed at a high level and miR-150-5p was downregulated in SDH of SNL rats. Exosomes injected were localized in rat SDH. BMSC-exosomes or NOTCH2 downregulation increased PWT and PWL of SNL rats and reduced apoptosis and inflammation in SDH. In contrast, NOTCH2 overexpression aggravated mechanical allodynia and SDH injury. Moreover, inhibiting miR-150-5p in BMSC-exosomes offset the therapeutic effects of BMSC-exosomes. Microglia activation induced mechanical allodynia in wild rats, while intrathecal injection of microglial cells incubated with BMSC-exosomes showed alleviated mechanical allodynia in SNL rats. NOTCH2 was targeted by miR-150-5p. Conclusion BMSC-derived exosomal miR-150-5p alleviates mechanical allodynia by targeting NOTCH2 in microglial cells.

Bibliometric and visual analysis of microglia-related neuropathic pain from 2000 to 2021

“…Intrathecal injection of MSC-EVs carrying miR-99b-3p reduced microglia activation in the spinal cord’s dorsal horn and mechanical allodynia in chronic constriction injury (CCI) rats by stimulating autophagy ( Gao et al, 2023 ). Li et al (2022) showed that the BMSC-derived exosomal miR-150-5p attenuates mechanical allodynia by targeting NOTCH2 in microglial. In addition to the beneficial action of MicroRNAs, Huc-MSCs-derived exosomes may exert analgesic effects on neuropathic pain by inhibiting activation of the TLR2/MyD88/NF-κB signaling pathway in spinal microglia ( Gao et al, 2023 ).…”

Stem cell-derived extracellular vesicle-based therapy for nerve injury: A review of the molecular mechanisms

“…Li et al documented that intrathecally administration of the bone marrow mesenchymal stem cell-derived exosomes loaded with miR-150–5p alleviate mechanical allodynia through targeting neurogenic locus notch homolog protein 2 (NOTCH2) in microglial cells following L5 spinal nerve ligation model in rat. 49 In this regard, another study showed that a single intrathecal injection of the human placental mesenchymal stem cells (hPMSCs)-derived small extracellular vesicles durably reversed mechanical hypersensitivity and neuroinflammation through miR-26a-5p/Wnt5a in mice with partial sciatic nerve ligation. 50 Histological and functional evaluation of the crushed sciatic nerve after transplanting the gel containing human multipotent mesenchymal stromal cell-derived extracellular vesicles (MSCs-EVs) showed a significant decrease in apoptotic neuronal death and an increase in regeneration-associated proteins NF-200 and GAP-43, as well as a decrease in muscle atrophy.…”

Clinical applications of stem cell-derived exosomes

“…These include, but are not limited to: 1, dementia, such as vascular dementia. 199 HIV-associated neurocognitive disorders 200 and radiation-induced cognitive dysfunction 201,202 ; 2, functional disorders, such as epilepsy 203 and mechanical allodynia 204 ; 3, congenital abnormalities, such as spina bifida aperta 205 ; 4, neuropsychological conditions, such as depression 206 and stress 207 ; 5, brain aging [208][209][210] ; 6, iatrogenic brain problems, such as deep hypothermic circulatory arrest. 211 PLASTIC SURGERY AND SC-EXO THERAPY Wound healing occurs in all parts of the human body, with cutaneous wounds being the most common.…”