MECHANISM OF MIR-25-3P CARRIED BY EXTRACELLULAR VESICLES DERIVED FROM PLATELET-RICH PLASMA IN IL-1β-INDUCED NUCLEUS PULPOSUS CELL DEGENERATION VIA THE SOX4/CXCR7 AXIS.

Shock (Augusta, Ga.) Pub Date : 2022-07-01 Epub Date: 2022-07-19 DOI:10.1097/SHK.0000000000001947
Baoshan Hu, Lianxin Wang, Naikun Sun, Shengrong Lin, Gang Rui
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Abstract

Abstract: Objectives: Nucleus pulposus (NP) cell degeneration promotes the progression of intervertebral disc (IVD) degeneration. MicroRNAs (miRs) are associated with IVD degeneration. This study expounded the mechanism of microRNA (miR)-25-3p carried by extracellular vesicles (EVs) derived from platelet-rich plasma (PRP) in interleukin (IL)-1β-induced NP cell degeneration. Methods: Platelet-rich plasma from mouse blood was obtained, and EVs were isolated from PRP (EVs derived from PRP [PRP-EVs]) and identified. Nucleus pulposus cells were isolated from the mouse lumbar IVD and treated with IL-1β to induce NP cell degeneration. Extracellular vesicles derived from PRP were added into NP cell culture medium. Afterward, intracellular miR-25-3p, sex determining region Y-related high-mobility-group box 4 (SOX4), and CXC chemokine receptor 7 (CXCR7) levels were examined. Nucleus pulposus cell viability, apoptosis, and inflammation were detected. Extracellular vesicles derived from PRP were labeled by PKH67 to obverse the uptake of EVs by NP cells. The binding relations between SOX4 and miR-25-3p and CXCR7 were predicted and examined. Functional rescue experiments were performed to investigate the roles of miR-25-3p, SOX4, and CXCR7 in NP cell degeneration. Results: miR-25-3p was downregulated, whereas SOX4 and CXCR7 were upregulated in IL-1β-induced NP cells. Extracellular vesicles derived from PRP increased the cell viability, and decreased apoptosis and inflammation. miR-25-3p carried by PRP-EVs into NP cells alleviated NP cell degeneration. miR-25-3p inhibited SOX4 expression and limited CXCR7 transcription. Silencing miR-25-3p or overexpressing SOX4 or CXCR7 reversed the alleviating role of PRP-EVs in NP cell degeneration. Conclusion: miR-25-3p carried by PRP-EVs into NP cells elevated intracellular miR-25-3p expression, which suppressed SOX4 expression and further limited CXCR7 transcription, thus alleviating IL-1β-induced NP cell degeneration. Extracellular vesicles derived from PRP containing miR-25-3p may be a new method for IVD treatment.

富血小板血浆细胞外囊泡携带mir-25-3p通过sox4 / cxcr7轴参与il -1β诱导的髓核细胞变性的机制。
摘要:目的:髓核(NP)细胞退变促进椎间盘退变的进展。MicroRNAs (miRs)与IVD变性相关。本研究阐述了富血小板血浆(PRP)来源的细胞外囊泡(ev)携带的microRNA (miR)-25-3p在白细胞介素(IL)-1β诱导的NP细胞变性中的作用机制。方法:从小鼠血液中获得富血小板血浆,从PRP中分离EVs (EVs derived from PRP [PRP-EVs])并进行鉴定。从小鼠腰椎IVD中分离髓核细胞,用IL-1β诱导NP细胞变性。将PRP衍生的细胞外囊泡加入NP细胞培养基中。随后,检测细胞内miR-25-3p、性别决定区y相关高流动性组盒4 (SOX4)和CXC趋化因子受体7 (CXCR7)水平。检测髓核细胞活力、凋亡及炎症反应。PRP衍生的细胞外囊泡被PKH67标记,以阻止NP细胞摄取ev。预测并检测SOX4与miR-25-3p和CXCR7的结合关系。通过功能修复实验研究miR-25-3p、SOX4和CXCR7在NP细胞变性中的作用。结果:miR-25-3p在il -1β诱导的NP细胞中下调,而SOX4和CXCR7上调。PRP衍生的细胞外囊泡增加了细胞活力,减少了细胞凋亡和炎症。prp - ev携带miR-25-3p进入NP细胞可减轻NP细胞的变性。miR-25-3p抑制SOX4表达,限制CXCR7转录。沉默miR-25-3p或过表达SOX4或CXCR7逆转了prp - ev在NP细胞变性中的缓解作用。结论:prp - ev携带miR-25-3p进入NP细胞,提高细胞内miR-25-3p的表达,抑制SOX4的表达,进一步限制CXCR7的转录,从而缓解il -1β诱导的NP细胞变性。含有miR-25-3p的PRP衍生的细胞外囊泡可能是治疗IVD的新方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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