中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (10): 1492-1500.doi: 10.12307/2023.270

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

miR-1-5p修饰脐带间充质干细胞治疗MRL/lpr狼疮小鼠的作用

丁丽丽1,胡明智1,武志慧1,孙晓林2,王永福1   

  1. 包头医学院第一附属医院,1风湿免疫科,2中心实验室(内蒙古自治区自体免疫学重点实验室),内蒙古自治区包头市   014010
  • 收稿日期:2022-02-10 接受日期:2022-04-29 出版日期:2023-04-08 发布日期:2022-09-08
  • 通讯作者: 王永福,博士,教授,包头医学院第一附属医院风湿免疫科,内蒙古自治区包头市 014010 孙晓林,博士,主管检验师,包头医学院第一附属医院中心实验室(内蒙古自治区自体免疫学重点实验室),内蒙古自治区包头市 014010
  • 作者简介:丁丽丽,女,1992年生,包头医学院在读硕士,主要从事自身免疫性疾病的研究。
  • 基金资助:
    国家自然科学基金项目(81860294),项目负责人:孙晓林;内蒙古自治区自然科学基金项目(2019MS08055),项目参与人:孙晓林;内蒙古自治区自然科学基金项目(2021MS08045),项目负责人:孙晓林;内蒙古自治区科技计划项目(201802089),项目参与人:孙晓林;内蒙古自治区科技计划项目(2019GG052),项目负责人:王永福

Therapeutic effect of umbilical cord mesenchymal stem cells modified by miR-1-5p in MRL/lpr lupus mice

Ding Lili1, Hu Mingzhi1, Wu Zhihui1, Sun Xiaolin2, Wang Yongfu1   

  1. 1Department of Rheumatology and Immunology, First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China; 2Central Laboratory (Key Laboratory of Autoimmunity in Inner Mongolia Autonomous Region), First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China
  • Received:2022-02-10 Accepted:2022-04-29 Online:2023-04-08 Published:2022-09-08
  • Contact: Wang Yongfu, MD, Professor, Department of Rheumatology and Immunology, First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China Sun Xiaolin, MD, Inspector-in-charge, Central Laboratory (Key Laboratory of Autoimmunity in Inner Mongolia Autonomous Region), First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China
  • About author:Ding Lili, Master candidate, Department of Rheumatology and Immunology, First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81860294 (to SXL); Natural Science Foundation of Inner Mongolia Autonomous Region, No. 2019MS08055 (to SXL); Natural Science Foundation of Inner Mongolia Autonomous Region, No. 2021MS08045 (to SXL); Science and Technology Planning Project of Inner Mongolia Autonomous Region, No. 201802089 (to SXL); Science and Technology Planning Project of Inner Mongolia Autonomous Region, No. 2019GG052 (to WYF)

摘要:

文题释义:
miR-1:是肌肉中特异性表达的miRNA,其可通过转录后水平参与基因表达的调节。目前已发现miR-1在多种疾病中存在异常表达,例如在多发性肌炎、皮肌炎和包涵体肌炎患者中均呈低表达。miR-1的表达下调,可抑制成肌细胞分化,参与到特发性炎症性肌病的发病过程中。
JAK/STAT信号通路:JAK/STAT作为细胞因子信号转导通路之一,参与多种自身免疫性疾病的调节。选择性的JAK抑制剂可降低克罗恩病患者C-反应蛋白水平,减轻炎症反应,促进损伤的肠黏膜愈合。在自身免疫性肝炎中,抑制JAK/STAT信号通路的激活,可减轻肝脏炎症,减少肝细胞凋亡。抑制JAK-STAT通路的激活可缓解系统性红斑狼疮相关脏器损伤。

背景:大量研究表明间充质干细胞能够通过多种免疫调节途径治疗自身免疫性疾病,基因修饰可能提高间充质干细胞的治疗潜力。miR-1-5p在系统性红斑狼疮中呈低表达,过表达miR-1-5p的间充质干细胞可能对系统性红斑狼疮具有治疗作用。
目的:探讨miR-1-5p修饰的脐带间充质干细胞是否介导JAK2/STAT3通路对系统性红斑狼疮 MRL/lpr模型小鼠存在治疗作用。
方法:将20只 MRL/lpr 狼疮小鼠随机分为miR-1-5p 转染脐带间充质干细胞治疗组(UC-MSCs+miR-1-5p组)、miRNA阴性对照转染脐带间充质干细胞治疗组(UC-MSCs+miR-NC组)、脐带间充质干细胞治疗组(UC-MSCs 组)、PBS治疗组(PBS组)、MRL/lpr狼疮小鼠未处理组(对照组)。以尾静脉移植的方式治疗MRL/lpr狼疮小鼠,每周1次,治疗5次。结束治疗后间隔1周处死小鼠,苏木精-伊红染色观察各组MRL/lpr小鼠肝组织和肠组织的病理变化;Western blot法检测各组小鼠肾组织JAK2/ STAT3通路蛋白及其磷酸化水平和白细胞介素18的表达水平。
结果与结论:①与对照组相比,UC-MSCs+miR-1-5p组、UC-MSCs+miR-NC组和UC-MSCs组MRL/lpr小鼠肝组织中炎性细胞浸润明显减少,UC-MSCs+miR-NC组可见肝细胞水肿变性;UC-MSCs+miR-1-5p组MRL/lpr小鼠肠组织中杯状细胞明显增多,其余各组差异不显著。②与对照组相比,UC-MSCs+miR-1-5p组狼疮小鼠肾组织中p-JAK2(Y1007+Y1008)、p-STAT3(s727)、白细胞介素18的表达水平显著下调(P < 0.05);UC-MSCs 组狼疮小鼠肾组织中p-STAT3(s727) (P < 0.05)、白细胞介素18 (P < 0.01)显著下调;UC-MSCs+ miR-NC组白细胞介素18的表达水平显著下调(P < 0.05);各组狼疮小鼠肾组织中p-STAT3(Y705)的表达差异无显著性意义(P > 0.05)。与PBS组相比,UC-MSCs 组白细胞介素18表达明显下调(P < 0.05)。③结果表明,miR-1-5p修饰的脐带间充质干细胞可能通过抑制JAK2/STAT3通路的激活和白细胞介素18的表达来缓解MRL/lpr小鼠肝肠损害,从而对狼疮小鼠起到一定的治疗作用。

https://orcid.org/0000-0002-2196-8008 (丁丽丽) 

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: miR-1-5p, 脐带间充质干细胞, 系统性红斑狼疮, JAK2/STAT3信号通路, 白细胞介素18

Abstract: BACKGROUND: Numerous studies have shown that mesenchymal stem cells can treat autoimmune diseases through a variety of immunomodulatory pathways, and genetic modification may improve the therapeutic potential of mesenchymal stem cells. MiR-1-5p is lowly expressed in systemic lupus erythematosus, and mesenchymal stem cells overexpressing miR-1-5p may have a therapeutic effect on systemic lupus erythematosus.  
OBJECTIVE: To investigate whether miR-1-5p-modified umbilical cord mesenchymal stem cells mediate the JAK2/STAT3 pathway and have a therapeutic effect on systemic lupus erythematosus MRL/lpr model mice.
METHODS:  Twenty MRL/lpr lupus mice were randomly divided into miR-1-5p transfected umbilical cord mesenchymal stem cells treatment group (UC-MSCs+miR-1-5p group), miRNA negative control transfected umbilical cord mesenchymal stem cells treatment group (UC-MSCs+miR-NC group), umbilical cord mesenchymal stem cell treatment group (UC-MSCs group), PBS treatment group (PBS group), and MRL/lpr lupus mice untreated (control group). MRL/lpr lupus mice were treated with tail vein transplantation, once a week, 5 times. Mice were sacrificed at 1-week intervals after the end of treatment. Hematoxylin-eosin staining was used to observe the pathological changes of liver tissue and intestinal tissue of MRL/lpr mice in each group. Western blot assay was used to detect JAK2/STAT3 pathway proteins and their phosphorylation levels and expression levels of interleukin-18 in kidney tissue of mice in each group.  
RESULTS AND CONCLUSION: (1) Compared with control group, the infiltration of inflammatory cells in liver tissue of MRL/lpr mice in UC-MSCs+miR-1-5p group, UC-MSCs+miR-NC group, and UC-MSCs group decreased significantly, and hepatocyte edema and degeneration could be seen in UC-MSCs+miR-NC group. Goblet cells in intestinal tissue of MRL/lpr mice in UC-MSCs+miR-1-5p group increased significantly, but there was no significant difference among other groups. (2) Compared with control group, the expression of p-JAK2 (Y1007+Y1008), p-STAT3 (s727) and interleukin-18 in renal tissue of lupus mice in UC-MSCs+miR-1-5p group was significantly down-regulated (P < 0.05); p-STAT3(s727) (P < 0.05) and interleukin-18 (P < 0.01) were significantly down-regulated in the kidney tissue of lupus mice in UC-MSCs group; expression level of interleukin-18 in UC-MSCs+miR-NC group was significantly down-regulated (P < 0.05); and there was no significant difference in the expression of p-STAT3 (Y705) in renal tissue of lupus mice of each group (P > 0.05). The expression of interleukin-18 in UC-MSCs group was significantly lower than that in PBS group (P < 0.05). (3) The results suggested that umbilical cord mesenchymal stem cells modified by miR-1-5p may alleviate liver and intestinal damage in MRL/lpr mice by inhibiting the activation of JAK2/STAT3 pathway and the expression of interleukin 18, thereby playing a certain therapeutic role in lupus mice.

Key words: miR-1-5p, umbilical cord-mesenchymal stem cell, systemic lupus erythematosus, JAK2/STAT3 signaling pathway, interleukin-18

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