Bone marrow mesenchymal stem cells-derived exosomal miR-381 alleviates lung ischemia-reperfusion injury by activating Treg differentiation through inhibiting YTHDF1 expression

IF 4.4 2区生物学 Q2 CELL BIOLOGY

Cellular signalling Pub Date : 2024-09-30 DOI:10.1016/j.cellsig.2024.111440

Cao Gao , Lei Chen , Xiang-yu Xie , Xiao-feng He , Jiang Shen , Liang Zheng

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Abstract

Aim

Our study aimed to investigate whether BMSCs-derived exosomal miR-381 promotes Treg cell differentiation in lung ischemia-reperfusion injury (LIRI), and the underlying mechanism.

Methods

The in vitro and in vivo models of LIRI were established by hypoxia/reoxygenation (H/R) treatment and lung ischemia/reperfusion (I/R) surgery, respectively. BMSCs-derived exosomes were isolated and identified by western blot, nanoparticle tracking analysis, and transmission electron microscopy. Cell viability, proliferation, and apoptosis were assessed by CCK-8, EdU, and flow cytometry assay, respectively. IL-18 secretion level in lung microvascular endothelial cells (LMECs) and lung tissue homogenate was examined by ELISA. Treg cell differentiation was determined using flow cytometry. The relationships between miR-381, YTHDF1, and IL-18 were investigated using dual-luciferase reporter gene, RIP, and/or RNA pull-down assays. MeRIP assay was employed to determine m⁶A modification of IL-18 mRNA in LMECs. The ubiquitination level of Foxp3 protein in CD4⁺ T cells was analyzed by Co-IP assay.

Results

BMSCs-derived exosomes reduced LMECs injury and increased Treg cell differentiation in LIRI, whereas miR-381 inhibition in BMSCs weakened these impacts. Mechanistically, miR-381 inhibited IL-18 translation in LMECs by inhibiting YTHDF1 expression via binding to its 3’-UTR. As expected, YTHDF1 overexpression in LMECs abolished the effects of miR-381-overexpressed exosomes on LMECs injury and Treg cell differentiation. Moreover, LMECs-secreted IL-18 inhibited Treg cell differentiation by promoting the ubiquitination degradation of Foxp3 protein.

Conclusion

BMSCs-derived exosomal miR-381 suppressed IL-18 translation in LMECs through binding to YTHDF1 3’-UTR, thus suppressing the ubiquitination degradation of Foxp3 in CD4⁺ T cells, which promoted Treg cell differentiation and mitigated LIRI development.

Abstract Image

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骨髓间充质干细胞衍生的外泌体miR-381通过抑制YTHDF1的表达激活Treg分化，从而减轻肺缺血再灌注损伤。

目的：我们的研究旨在探讨BMSCs衍生的外泌体miR-381是否能促进肺缺血再灌注损伤（LIRI）中Treg细胞的分化及其内在机制：分别通过缺氧/再氧合（H/R）治疗和肺缺血/再灌注（I/R）手术建立肺缺血再灌注损伤的体外和体内模型。通过Western印迹、纳米颗粒追踪分析和透射电子显微镜分离并鉴定了BMSCs衍生的外泌体。细胞活力、增殖和凋亡分别通过 CCK-8、EdU 和流式细胞术进行评估。用 ELISA 检测肺微血管内皮细胞（LMECs）和肺组织匀浆中 IL-18 的分泌水平。流式细胞术测定了 Treg 细胞的分化情况。使用双荧光素酶报告基因、RIP 和/或 RNA 拉取试验研究了 miR-381、YTHDF1 和 IL-18 之间的关系。MeRIP 分析法用于确定 LMECs 中 IL-18 mRNA 的 m6A 修饰。Co-IP 试验分析了 CD4+ T 细胞中 Foxp3 蛋白的泛素化水平：结果：BMSCs衍生的外泌体减轻了LIRI中LMECs的损伤并增加了Treg细胞的分化，而抑制BMSCs中的miR-381则削弱了这些影响。从机理上讲，miR-381 通过与其 3'-UTR 结合抑制 YTHDF1 的表达，从而抑制了 LMECs 中 IL-18 的翻译。不出所料，LMECs 中 YTHDF1 的过表达消除了 miR-381 表达的外泌体对 LMECs 损伤和 Treg 细胞分化的影响。此外，LMECs分泌的IL-18通过促进Foxp3蛋白的泛素化降解抑制了Treg细胞的分化：结论：BMSCs衍生的外泌体miR-381通过与YTHDF1 3'-UTR结合，抑制了LMECs中IL-18的翻译，从而抑制了CD4+ T细胞中Foxp3的泛素化降解，促进了Treg细胞的分化，缓解了LIRI的发展。

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来源期刊

Cellular signalling 生物-细胞生物学

CiteScore

8.40

自引率

0.00%

发文量

250

审稿时长

27 days

期刊介绍： Cellular Signalling publishes original research describing fundamental and clinical findings on the mechanisms, actions and structural components of cellular signalling systems in vitro and in vivo. Cellular Signalling aims at full length research papers defining signalling systems ranging from microorganisms to cells, tissues and higher organisms.