miR-125b-5p修饰脐带间充质干细胞对系统性红斑狼疮的免疫调控机制

武志慧; 胡明智; 赵巧英; 吕凤凤; 张晶莹; 张伟; 王永福; 孙晓林; 王慧

doi:10.19723/j.issn.1671-167X.2024.05.017

北京大学学报（医学版） >

2024 , Vol. 56 >Issue 5: 860 - 867

DOI: https://doi.org/10.19723/j.issn.1671-167X.2024.05.017

论著

miR-125b-5p修饰脐带间充质干细胞对系统性红斑狼疮的免疫调控机制

武志慧 ,
胡明智 ,
赵巧英 ,
吕凤凤 ,
张晶莹 ,
张伟 ,
王永福 ,
孙晓林 ,
王慧

展开

1. 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 014010
2. 包头医学院第一附属医院风湿免疫科，内蒙古自治区包头 014010

收稿日期: 2021-07-26

网络出版日期: 2024-10-16

基金资助

国家自然科学基金(81860294);国家自然科学基金(81860295);内蒙古自治区自然科学基金(2019MS08055);内蒙古自治区科技计划项目(201802089);内蒙古自治区科技计划项目(2019GG052)

版权

收起

Immunomodulatory mechanism of umbilical cord mesenchymal stem cells modified by miR-125b-5p in systemic lupus erythematosus

Zhihui WU ,
Mingzhi HU ,
Qiaoying ZHAO ,
Fengfeng LV ,
Jingying ZHANG ,
Wei ZHANG ,
Yongfu WANG ,
Xiaolin SUN ,
Hui WANG

Expand

1. Central Laboratory, First Affiliated Hospital of Baotou Medical College (Inner Mongolia Key Laboratory of Autoimmunology), Baotou 014010, Inner Mongolia Autonomous Region, China
2. Department of Rheumatism and Immunology, First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China

SUN Xiaolin, e-mail, yolo_1010@126.com
WANG Hui, e-mail, wanghuier@126.com

Received date: 2021-07-26

Online published: 2024-10-16

Supported by

Supported by the National Natural Science Foundation of China(81860294);Supported by the National Natural Science Foundation of China(81860295);the Natural Science Foundation of Inner Mongolia(2019MS08055);the Inner Mongolia Autonomous Region Science and Technology Plan Projects(201802089);the Inner Mongolia Autonomous Region Science and Technology Plan Projects(2019GG052)

Copyright

Fold

摘要

目的: 研究miR-125b-5p修饰脐带间充质干细胞(umbilical cord mesenchymal stem cells，UC-MSCs)对系统性红斑狼疮(systemic lupus erythematosus，SLE)的免疫调控作用机制。方法: 实时荧光定量PCR检测miR-125b-5p在UC-MSCs和SLE患者、健康体检者外周血单个核细胞中的表达水平；Annexin V-FITC/PI凋亡检测试剂盒检测miR-125b-5p对UC-MSCs的凋亡影响；每周对MRL/lpr小鼠进行尾静脉注射UC-MSCs，5周后流式细胞术检测各组小鼠脾细胞T淋巴细胞亚群分化情况；ELISA法检测各组MRL/lpr小鼠血清中白细胞介素(interleukin，IL)-4和IL-17A表达水平；苏木精-伊红染色法观察MRL/lpr小鼠肺和肾组织病理变化。结果: 与健康对照组相比，SLE患者外周血单个核细胞中miR-125b-5p的表达水平显著下调(P < 0.01)；与UC-MSCs相比，miR-125b-5p转染UC-MSCs组miR-125b-5p的表达水平显著上调(P < 0.01)。miR-125b-5p转染UC-MSCs 48 h可显著提高细胞存活率(P < 0.01)；与未处理的MRL/lpr小鼠组相比，miR-125b-5p修饰UC-MSCs对MRL/lpr小鼠脾脏Th17细胞分化具有明显的下调作用(P < 0.05)，且在小鼠血清中IL-4表达水平明显升高(P < 0.05)，IL-17A表达水平明显减低(P < 0.05)。经miR-125b-5p修饰的UC-MSCs治疗后，MRL/lpr小鼠肺和肾组织内炎性细胞浸润和微血栓减少。结论: miR-125b-5p修饰UC-MSCs对SLE具有免疫调控作用。

关键词： 间充质干细胞; 基因修饰; 系统性红斑狼疮; 免疫调节; 脐带

本文引用格式

武志慧 , 胡明智 , 赵巧英 , 吕凤凤 , 张晶莹 , 张伟 , 王永福 , 孙晓林 , 王慧 . miR-125b-5p修饰脐带间充质干细胞对系统性红斑狼疮的免疫调控机制[J]. 北京大学学报（医学版）, 2024 , 56(5) : 860 -867 . DOI: 10.19723/j.issn.1671-167X.2024.05.017

Abstract

Objective: To investigate the mechanism of immunomodulatory effects of umbilical cord mesenchymal stem cells (UC-MSCs) modified by miR-125b-5p on systemic lupus erythematosus (SLE). Methods: The expression level of miR-125b-5p was detected by real-time fluorescence quantitative PCR in UC-MSCs and peripheral blood mononuclear cells (PBMCs) from SLE patients and health checkers. Annexin V-FITC/PI apoptosis detection kit was used to detect the effect of miR-125b-5p on apoptosis of UC-MSCs. MRL/lpr mice in each group were injected with UC-MSCs via tail vein, and T-lymphocyte subsets in the spleen of the MRL/lpr mice were detected by flow cytometry after 5 weeks. The expression levels of interleukin (IL)-4 and IL-17A in serum of MRL/lpr mice were detected by ELISA. Hematoxylin-eosin staining was used to observe the pathological manifestations of the lungs and kidneys of the MRL/lpr mice. Results: miR-125b-5p was significantly down-regulated in PBMCs of SLE patients compared with healthy controls (P < 0.01). Compared with the UC-MSCs group, the expression of miR- 125b-5p in UC-MSCs modified by miR-125b-5p group was increased (P < 0.01). The survival rate of UC-MSCs was significantly increased by miR-125b-5p (P < 0.01). Compared with the untreated group of MRL/lpr mice, the expression level of IL-4 in serum was increased (P < 0.05); the expression level of IL-17A was decreased (P < 0.05); the proportion of Th17 cells in the spleen of MRL/lpr mice was decreased (P < 0.05); the inflammatory cells infiltration and micro-thrombosis of lungs and kidneys of MRL/lpr mice were significantly reduced in the UC-MSCs modified by miR-125b-5p treatment group. Conclusion: UC-MSCs modified by miR-125b-5p have immunomodulatory effects on systemic lupus erythematosus.

Key words： Mesenchymal stem cells; Gene modification; Systemic lupus erythematosus; Immunomo-dulation; Umbilical cord

参考文献

1	Tsokos GC . Systemic lupus erythematosus[J]. N Engl J Med, 2011, 365 (22): 2110- 2121.
2	Relle M , Foehr B , Schwarting A . Epigenetic aspects of systemic lupus erythematosus[J]. Rheumatol Ther, 2015, 2 (1): 33- 46.
3	Basta F , Fasola F , Triantafyllias K , et al. Systemic lupus erythematosus (SLE) therapy: The old and the new[J]. Rheumatol Ther, 2020, 7 (3): 433- 446.
4	Sharabi A , Tsokos GC . T cell metabolism: New insights in systemic lupus erythematosus pathogenesis and therapy[J]. Nat Rev Rheumatol, 2020, 16 (2): 100- 112.
5	Muhammad Yusoff F , Wong KK , Mohd Redzwan N . Th1, Th2, and Th17 cytokines in systemic lupus erythematosus[J]. Autoimmunity, 2020, 53 (1): 8- 20.
6	Serakinci N , Fahrioglu U , Christensen R . Mesenchymal stem cells, cancer challenges and new directions[J]. Eur J Cancer, 2014, 50 (8): 1522- 1530.
7	Drela K , Lech W , Figiel-Dabrowska A , et al. Enhanced neuro-therapeutic potential of Wharton' s jelly-derived mesenchymal stem cells in comparison with bone marrow mesenchymal stem cells culture[J]. Cytotherapy, 2016, 18 (4): 497- 509.
8	Liu L , Wong CW , Han M , et al. Meta-analysis of preclinical studies of mesenchymal stromal cells to treat rheumatoid arthritis[J]. EBioMedicine, 2019, 47, 563- 577.
9	Qi J , Tang X , Li W , et al. Mesenchymal stem cells inhibited the differentiation of MDSCs via COX2/PGE2 in experimental sialadenitis[J]. Stem Cell Res Ther, 2020, 11 (1): 325.
10	Liu C , Zhang H , Tang X , et al. Mesenchymal stem cells promote the osteogenesis in collagen-induced arthritic mice through the inhibition of TNF-α[J]. Stem Cells Int, 2018, 2018, 4069032.
11	Choudhery MS , Badowski M , Muise A , et al. Donor age negatively impacts adipose tissue-derived mesenchymal stem cell expansion and differentiation[J]. J Transl Med, 2014, 12, 8.
12	Escacena N , Quesada-Hernández E , Capilla-Gonzalez V , et al. Bottlenecks in the efficient use of advanced therapy medicinal products based on mesenchymal stromal cells[J]. Stem Cells Int, 2015, 2015, 895714.
13	Fischer UM , Harting MT , Jimenez F , et al. Pulmonary passage is a major obstacle for intravenous stem cell delivery: The pulmonary first-pass effect[J]. Stem Cells Dev, 2009, 18 (5): 683- 692.
14	Abdelmohsen K , Gorospe M . Noncoding RNA control of cellular senescence[J]. Wiley Interdiscip Rev RNA, 2015, 6 (6): 615- 629.
15	Su T , Xiao Y , Xiao Y , et al. Bone marrow mesenchymal stem cells-derived exosomal MiR-29b-3p regulates aging-associated insulin resistance[J]. ACS Nano, 2019, 13 (2): 2450- 2462.
16	Meng Y , Eirin A , Zhu XY , et al. Micro-RNAs regulate metabolic syndrome-induced senescence in porcine adipose tissue-derived mesenchymal stem cells through the P16/MAPK pathway[J]. Cell Transplant, 2018, 27 (10): 1495- 1503.
17	Vishnoi A , Rani S . MiRNA biogenesis and regulation of diseases: An overview[J]. Methods Mol Biol, 2017, 1509, 1- 10.
18	Gong B , Zheng L , Lu Z , et al. Mesenchymal stem cells negatively regulate CD4⁺ T cell activation in patients with primary Sj?gren syndrome through the miRNA-125b and miRNA-155 TCR pathway[J]. Mol Med Rep, 2021, 23 (1): 43.
19	Xiu L , Xing Q , Mao J , et al. miRNA-125b-5p suppresses hypo-thyroidism development by targeting signal transducer and activator of transcription 3[J]. Med Sci Monit, 2018, 24, 5041- 5049.
20	胡明智, 张晶莹, 杨国安, 等. miR-1-5p修饰脐带间充质干细胞对系统性红斑狼疮T淋巴细胞亚群的免疫调节[J]. 中国组织工程研究, 2021, 25 (31): 4928- 4938.
21	Gentile P , Sterodimas A . Adipose-derived stromal stem cells (ASCs) as a new regenerative immediate therapy combating coronavirus (COVID-19)-induced pneumonia[J]. Expert Opin Biol Ther, 2020, 20 (7): 711- 716.
22	Toyserkani NM , J?rgensen MG , Tabatabaeifar S , et al. Concise review: A safety assessment of adipose-derived cell therapy in cli-nical trials: A systematic review of reported adverse events[J]. Stem Cells Transl Med, 2017, 6 (9): 1786- 1794.
23	Chen C , Liang J , Yao G , et al. Mesenchymal stem cells upregulate Treg cells via sHLA-G in SLE patients[J]. Int Immuno-pharmacol, 2017, 44, 234- 241.
24	张立民. 系统性红斑狼疮microRNA表达谱和功能的初步研究[D]. 北京: 中国协和医科大学, 2010.
25	Wang D , Huang S , Yuan X , et al. The regulation of the Treg/Th17 balance by mesenchymal stem cells in human systemic lupus erythematosus[J]. Mol Immunol, 2017, 14 (5): 423- 431.
26	Golpanian S , DiFede DL , Pujol MV , et al. Rationale and design of the allogeneiC human mesenchymal stem cells (hMSC) in patients with aging fRAilTy via intravenoUS delivery (CRATUS) study: A phase Ⅰ/Ⅱ, randomized, blinded and placebo controlled trial to evaluate the safety and potential efficacy of allogeneic human mesenchymal stem cell infusion in patients with aging frailty[J]. Oncotarget, 2016, 7 (11): 11899- 11912.
27	Yang J , Yang X , Zou H , et al. Oxidative stress and Treg and Th17 dysfunction in systemic lupus erythematosus[J]. Oxid Med Cell Longev, 2016, 2016, 2526174.
28	Li D , Guo B , Wu H , et al. Interleukin-17 in systemic lupus erythematosus: A comprehensive review[J]. Autoimmunity, 2015, 48 (6): 353- 361.
29	Chen DY , Chen YM , Wen MC , et al. The potential role of Th17 cells and Th17-related cytokines in the pathogenesis of lupus nephritis[J]. Lupus, 2012, 21 (13): 1385- 1396.
30	La Cava A . Tregs in SLE: An Update[J]. Curr Rheumatol Rep, 2018, 20 (2): 6.
31	Beringer A , Noack M , Miossec P . IL-17 in chronic inflammation: From discovery to targeting[J]. Trends Mol Med, 2016, 22 (3): 230- 241.
32	Dolff S , Bijl M , Huitema MG , et al. Disturbed Th1, Th2, Th17 and T(reg) balance in patients with systemic lupus erythematosus[J]. Clin Immunol, 2011, 141 (2): 197- 204.
33	Moseley TA , Haudenschild DR , Rose L , et al. Interleukin-17 family and IL-17 receptors[J]. Cytokine Growth Factor Rev, 2003, 14 (2): 155- 174.
34	Qu N , Xu M , Mizoguchi I , et al. Pivotal roles of T-helper 17-related cytokines, IL-17, IL-22, and IL-23, in inflammatory diseases[J]. Clin Dev Immunol, 2013, 2013, 968549.
35	Ghali JR , Holdsworth SR , Kitching AR . Targeting IL-17 and IL-23 in immune mediated renal disease[J]. Curr Med Chem, 2015, 22 (38): 4341- 4365.
36	Talaat RM , Mohamed SF , Bassyouni IH , et al. Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus (SLE) patients: Correlation with disease activity[J]. Cytokine, 2015, 72 (2): 146- 153.
37	Zickert A , Amoudruz P , Sundstr?m Y , et al. IL-17 and IL-23 in lupus nephritis: Association to histopathology and response to treatment[J]. BMC immunol, 2015, 16 (1): 7.
38	Jha AN , Singh VK , Kumari N , et al. IL-4 haplotype -590T, -34T and intron-3 VNTR R2 is associated with reduced malaria risk among ancestral indian tribal populations[J]. PLoS One, 2012, 7 (10): e48136.
39	Patterson D , Jones C , Hart I , et al. The human interleukin-1 receptor antagonist (IL1RN) gene is located in the chromosome 2q14 region[J]. Genomics, 1993, 15 (1): 173- 176.
40	Rapoport M , Bloch O . Systemic lupus erythematosus[J]. N Engl J Med, 2012, 366 (6): 574.
41	Tsokos GC , Lo MS , Costa Reis P , et al. New insights into the immunopathogenesis of systemic lupus erythematosus[J]. Nat Rev Rheumatol, 2016, 12 (12): 716- 730.

Options

文章导航

模态框（Modal）标题

摘要

本文引用格式

Abstract

参考文献