中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (13): 2706-2712.doi: 10.12307/2025.048

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐带间充质干细胞来源外泌体抗小鼠肾缺血再灌注损伤的机制

李灵玉1，魏华锋1，罗  豪1，王  浩2，贺佳辉1，姚雅韡1，吕兴华2   

  1. 1兰州大学第一临床医学院，甘肃省兰州市   730000；2兰州大学第一医院日间手术中心，甘肃省兰州市   730000
  • 收稿日期:2024-01-05 接受日期:2024-03-15 出版日期:2025-05-08 发布日期:2024-09-11
  • 通讯作者: 吕兴华，博士，硕士生导师，主任医师，兰州大学第一医院日间手术中心，甘肃省兰州市 730000
  • 作者简介:李灵玉，女，1996年生，宁夏回族自治区吴忠市人，回族，兰州大学在读硕士，主要从事干细胞外泌体、肾缺血再灌注研究。
  • 基金资助:
    甘肃省自然科学基金(23JRRA0932)，项目负责人：吕兴华；甘肃省重点人才项目(2021RCXM002)，项目负责人：吕兴华；兰州大学第一医院院内基金(ldyyyn2020-106，ldyyyn2022-1)，项目负责人：吕兴华；兰州大学医学教育创新发展项目(lzuyxcx-2022-130)，项目负责人：吕兴华

Mechanism of human umbilical cord mesenchymal stem cell-derived exosomes against mouse renal ischemia/reperfusion injury

Li Lingyu1, Wei Huafeng1, Luo Hao1, Wang Hao2, He Jiahui1, Yao Yawei1, Lyu Xinghua2   

  1. 1First Clinical Medical College of Lanzhou University, Lanzhou 730000, Gansu Province, China; 2Day Surgery Center, The First Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China
  • Received:2024-01-05 Accepted:2024-03-15 Online:2025-05-08 Published:2024-09-11
  • Contact: Lyu Xinghua, MD, Master’s supervisor, Chief physician, Day Surgery Center, The First Hospital of Lanzhou University, Lanzhou 730000, Gansu Province, China
  • About author:Li Lingyu, Master candidate, First Clinical Medical College of Lanzhou University, Lanzhou 730000, Gansu Province, China
  • Supported by:
    Natural Science Foundation of Gansu Province, No. 23JRRA0932 (to LXH); Key Talent Project of Gansu Province, No. 2021RCXM002 (to LXH); Internal Hospital Fund of The First Hospital of Lanzhou University, Nos. ldyyyn2020-106, ldyyyn2022-1 (to LXH); Medical Education Innovation and Development Project of Lanzhou University, No. lzuyxcx-2022-130 (to LXH)

PDF

240

摘要：

文题释义：

人脐带间充质干细胞：来源于新生儿脐带组织中的一种多功能干细胞，因能分化为间充质组织而得名，具有亚全能分化潜能，在特定的体内、体外环境中能够诱导分化成为多种组织细胞。
外泌体：多囊泡体与细胞膜融合后释放入细胞间隙的直径为40-160 μm的细胞外囊泡，富含多种mRNA和蛋白质，通过分泌、摄取等相互作用参与细胞间通讯。

摘要
背景：人脐带间充质干细胞来源外泌体参与多种损伤修复过程，其对肾缺血再灌注损伤的影响以及具体机制还没有完全阐明。
目的：探讨人脐带间充质干细胞来源外泌体治疗肾缺血再灌注损伤的分子机制。
方法：①培养人脐带间充质干细胞，使用外泌体提取试剂盒获取外泌体并鉴定；②采用活体荧光成像技术检测外泌体在肾缺血再灌注损伤小鼠肾脏中的分布；③C57/BL6雄性小鼠30只，按随机数字表法分为5组：假手术组、肾缺血再灌注组、假手术组+Compound C组、肾缺血再灌注+外泌体组、肾缺血再灌注+外泌体+Compound C组，每组 6 只。除假手术组外，其余组均夹闭双侧肾蒂45 min，再灌注24 h建立肾缺血再灌注小鼠模型；假手术组+Compound C组和外泌体+Compound C组于造模前30 min腹腔注射AMPK抑制剂Compound C；外泌体组和外泌体+Compound C组在肾蒂夹闭前15 min经尾静脉注射外泌体。再灌注24 h检测各组小鼠血清肌酐和尿素氮水平、肾组织白细胞介素6和肿瘤坏死因子α水平、肾组织凋亡相关因子的表达。
结果与结论：①人脐带间充质干细胞外泌体具有典型的茶托形态，直径分布在40-160 nm范围内，表达外泌体表面特异性标志膜蛋白；②与假手术组相比，肾缺血再灌注损伤小鼠肾脏更易聚集人脐带间充质干细胞外泌体；③外泌体预处理可减轻肾缺血再灌注小鼠肾损伤，降低肾小管上皮细胞凋亡水平，并且这种保护作用可被AMPK抑制剂所逆转。结果表明，人脐带间充质干细胞来源外泌体发挥肾缺血再灌注损伤的保护作用可能与激活AMPK/YAP1通路抗细胞凋亡有关。

https://orcid.org/0000-0003-3938-3856 (李灵玉)；https://orcid.org/0000-0002-6757-6974 (吕兴华)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 人脐带间充质干细胞, 外泌体, AMPK-YAP1通路, 凋亡, 肾, 缺血再灌注损伤

Abstract: BACKGROUND: Human umbilical cord mesenchymal stem cell-derived exosomes are involved in multiple injury repair processes, and the effects and the specific mechanisms of renal ischemia/reperfusion injury have not been fully elucidated.
OBJECTIVE: To investigate the molecular mechanism of human umbilical cord mesenchymal stem cell-derived exosomes in the treatment of renal ischemia/reperfusion injury.
METHODS: (1) Human umbilical cord mesenchymal stem cells were cultured and exosomes were obtained and identified using an exosome extraction kit. (2) The distribution of exosomes in the kidney of mice with renal ischemia/reperfusion injury was examined by intravital fluorescence imaging. (3) Thirty C57/BL6 male mice were divided into five groups according to the random number table method: sham operation group, renal ischemia/reperfusion group, sham operation group+Compound C group, renal ischemia/reperfusion+exosome group (exosome group), and renal ischemia/reperfusion+exosome+Compound C group (exosome+Compound C group), with 6 mice in each group. Except the sham operation group, bilateral renal pedicles were clamped for 45 minutes and a mouse model of renal ischemia/reperfusion injury was established after 24 hours of reperfusion. In sham operation+Compound C group and exosome+Compound C group, AMPK inhibitor Compound C was intraperitoneally injected 30 minutes before model establishment. In the exosome group and exosome+Copmpound C group, exosomes were injected through the tail vein 15 minutes before renal pedicle clipping. The levels of serum creatinine and urea nitrogen, interleukin 6, and tumor necrosis factor α in renal tissue, and the expression of apoptosis-related factors in renal tissue were detected after 24 hours of reperfusion in each group. 
RESULTS AND CONCLUSION: (1) Human umbilical cord mesenchymal stem cell exosomes had the typical tea tray morphology, with the diameter distribution in the range of 40-160 nm, and expressed the specific marker membrane protein of exosome surface. (2) Murine kidneys after renal ischemia/reperfusion injury were more likely to gather human umbilical cord mesenchymal stem cell exosomes compared with the sham operation group. (3) Exosome pretreatment reduced renal injury and the level of renal cell apoptosis in mice treated with renal ischemia/reperfusion injury. Moreover, this protective effect could be reversed by AMPK inhibitors. These findings verify that human umbilical cord mesenchymal stem cell-derived exosomes exerting a protective effect on renal ischemia/reperfusion injury may be related to the activation of the AMPK/YAP1 pathway to antiapoptosis.

Key words: human umbilical cord mesenchymal stem cell, exosome, AMPK-YAP1 pathway, apoptosis, kidney, ischemia/reperfusion injury

中图分类号: