The Second Department of General Surgery, Yunnan University Affiliated Hospital, Kunming, Yunnan, P.R. China.
FASEB J. 2024 Dec 15;38(23):e70042. doi: 10.1096/fj.202400803RR.
As a common digestive disease, acute pancreatitis (AP) often threatens the life of patients. Bone marrow mesenchymal stem cells (BMSCs) derived exosomes have exhibited some benefits for AP. However, the mechanism remains unclear and deserves to be further investigated. The characteristics of BMSCs-exosomes (BMSCs-Exos) were identified. The abundance of genes and proteins was evaluated using quantitative real-time PCR (RT-qPCR), western blot, enzyme-linked immunosorbent assay (ELISA) and IF assay. Cell apoptosis and CD206-positive cells were measured by flow cytometry. The interactions among miR-181a-5p, Zinc finger E-box binding homeobox 2 (ZEB2) and Receptor for Activated C Kinase 1 (RACK1) were verified using dual luciferase reporter assay, RNA immunoprecipitation (RIP), coimmunoprecipitation (Co-IP). BMSCs-Exos effectively improved AP injury through restraining AR42J cell apoptosis and promoting M2 macrophage polarization, which was realized due to BMSCs-Exos harboring an abundance of miR-181a-5p. Further experiments validated miR-181a-5p silenced ZEB2 and ZEB2 reduced RACK1 expression through mediating RACK1 ubiquitination. ZEB2 knockdown decreased AR42J cell apoptosis and induced M2 macrophage polarization to alleviate AP injury, whereas RACK1 downregulation abolished these phenomena. BMSCs-Exos harboring miR-181a-5p suppressed AR42J cell apoptosis and promoted M2 macrophage polarization to delay AP progression through ZEB2-mediated RACK1 ubiquitination.
作为一种常见的消化系统疾病，急性胰腺炎（AP）常威胁患者生命。骨髓间充质干细胞（BMSCs）衍生的外泌体对 AP 具有一定的治疗作用。然而，其作用机制尚不清楚，值得进一步研究。本研究旨在鉴定 BMSCs 衍生外泌体（BMSCs-Exos）的特征，采用实时定量 PCR（RT-qPCR）、Western blot、酶联免疫吸附试验（ELISA）和免疫荧光检测法评估基因和蛋白丰度，通过流式细胞术检测细胞凋亡和 CD206 阳性细胞数，利用双荧光素酶报告基因检测、RNA 免疫沉淀（RIP）和免疫共沉淀（Co-IP）实验验证 miR-181a-5p、锌指 E 框结合同源盒 2（ZEB2）和激活蛋白激酶 C 受体 1（RACK1）之间的相互作用。结果表明，BMSCs-Exos 通过抑制 AR42J 细胞凋亡和促进 M2 型巨噬细胞极化来有效改善 AP 损伤，这是由于 BMSCs-Exos 富含 miR-181a-5p 所致。进一步的实验验证了 miR-181a-5p 通过沉默 ZEB2 并介导 RACK1 泛素化降低 RACK1 表达，ZEB2 敲低减少 AR42J 细胞凋亡并诱导 M2 型巨噬细胞极化从而减轻 AP 损伤，而 RACK1 下调则消除了这些现象。综上所述，BMSCs-Exos 可通过 ZEB2 介导的 RACK1 泛素化抑制 AR42J 细胞凋亡和促进 M2 型巨噬细胞极化来延缓 AP 进展。
