过表达<italic style="font-style: italic">miR</italic>-<italic style="font-style: italic">124</italic>-<italic style="font-style: italic">1</italic>基因的骨髓间充质干细胞外泌体调控小胶质细胞M2型极化对脑卒中的影响

郝磊; 卢柳西; 李琼莉; 孙洋; 秦翠玲; 展群岭

doi:10.16016/j.2097-0927.202305003

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过表达miR-124-1基因的骨髓间充质干细胞外泌体调控小胶质细胞M2型极化对脑卒中的影响

基础医学 | 更新时间：2026-01-09

- 过表达miR-124-1基因的骨髓间充质干细胞外泌体调控小胶质细胞M2型极化对脑卒中的影响
- Effect of exosomes derived from mesenchymal stem cells with miR-124-1 overexpression on rat model of stroke by regulating M2 polarization of microglia
- 陆军军医大学学报 2024年46卷第5期页码：458-466
- 作者机构：
  
  1.400062 重庆，重庆市第五人民医院神经内科
  2.400042 重庆，重庆医科大学附属第一医院第一分院神经内科
- 作者简介：
  
  [ 展群岭，E-mail：aczhan@163.com ZHAN Qunling, E-mail: aczhan@163.com
- 基金信息：
  
  重庆市自然科学基金面上项目(cstc2020jcyj-msxmX0988)
- DOI：10.16016/j.2097-0927.202305003
  中图分类号： R329.25;R394.3;R743.3
- 收稿：2023-05-04，
  
  修回：2023-07-13，
  
  纸质出版：2024-03-15
- 稿件说明：
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郝磊, 卢柳西, 李琼莉, 等. 过表达miR-124-1基因的骨髓间充质干细胞外泌体调控小胶质细胞M2型极化对脑卒中的影响[J]. 陆军军医大学学报, 2024,46(5):458-466.

Lei HAO, Liuxi LU, Qiongli LI, et al. Effect of exosomes derived from mesenchymal stem cells with miR-124-1 overexpression on rat model of stroke by regulating M2 polarization of microglia[J]. Journal of Army Medical University, 2024, 46(5): 458-466.
郝磊, 卢柳西, 李琼莉, 等. 过表达miR-124-1基因的骨髓间充质干细胞外泌体调控小胶质细胞M2型极化对脑卒中的影响[J]. 陆军军医大学学报, 2024,46(5):458-466. DOI： 10.16016/j.2097-0927.202305003.

Lei HAO, Liuxi LU, Qiongli LI, et al. Effect of exosomes derived from mesenchymal stem cells with miR-124-1 overexpression on rat model of stroke by regulating M2 polarization of microglia[J]. Journal of Army Medical University, 2024, 46(5): 458-466. DOI： 10.16016/j.2097-0927.202305003.

摘要

目的

探讨骨髓间充质干细胞(bone marrow derived mesenchymal stem cells，BMMSCs)外泌体(exosomes，Exo)过表达

miR

124

基因(Exo/124-1)调控小胶质细胞(microglia，MG)的M2型极化对脑卒中的影响。

方法

分离培养大鼠BMMSCs，收集其Exo(BMMSCs-Exo)，采用流式细胞术、Western blot与透射电子显微镜(transmission electron microscope，TEM)分别对其进行检测鉴定。30只大鼠简单随机分为假手术组(Sham组)、模型组(MCAO/R组)、Exo移植组(Exo组)、空病毒(Empty lentivirus，Elv)转染Exo移植组(Exo-Elv组)及Exo/124-1移植组(Exo/124-1组)，每组6只。Sham组仅行假手术，其余各组均复制大脑中动脉栓塞/再灌注(middle cerebral artery occlusion and reperfusion，MCAO/R)模型。模型复制1 d与14 d后，各移植组动物于右侧脑室植入相应移植物，Sham组、模型组注入相同剂量生理盐水作对照。术后2 h及1、3、7、14、21、28 d，分别对各组动物行改良神经系统严重程度评分(modified neurological severity scores，mNSS)。三苯基四氮唑(triphenyl tetrazolium chloride，TTC)染色检测其梗死体积。在基因与蛋白水平分别检测各组28 d脑组织MG的M1型分子[肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)]、M2型分子(CD206)的表达情况。

结果

成功获取并鉴定了BMMSCs及其Exo。Exo/124-1显著表达

miR

124

。所有动物(假手术组除外)术后出现神经功能缺损。术后7~28 d，Exo/124-1组的mNSS明显低于MCAO/R组(

＜0.05)与Exo组、Exo-Elv组(

＜0.01)；术后28 d，Exo/124-1组的脑梗死体积、TNF-α的表达明显小于MCAO/R组(

＜0.01)与Exo组、Exo-Elv组(

＜0.01)，CD206的表达显著高于MCAO/R组(

＜0.01)与Exo组、Exo-Elv组(

＜0.01)。

结论

BMMSCs-Exo携带

miR

124

基因可能调控MG的M2型极化，抑制M1型介导的炎症反应，促进脑卒中大鼠神经功能的恢复。

Abstract

Objective

To investigate the effect of exosomes (Exo) derived from bone marrow mesenchymal stem cells (BMMSCs) overexpressing

miR

124

by regulating M2 polarization of microglia (MG) on stroke.

Methods

Rat BMMSCs were isolated and cultured. Then BMMSCs-Exo were extracted

and flow cytometry

Western blotting and transmission electron microscopy (TEM) were used to identify the BMMSCs and BMMSCs-Exo

respectively. Thirty rats were randomized into sham operation group (sham group)

model group (middle cerebral artery occlusion and reperfusion

MCAO/R)

Exo transplantation group (Exo group)

transplantation group of Exo transfected with empty lentivirus (Exo-Elv group) and transplantation group of Exo transfected with124-1 lentivirus (Exo/124-1 group)

with 6 rats in each group. In 1 and 14 d after modeling

the corresponding agents were transplanted into the right lateral ventricle of rats in each transplantation group

while the rats of Sham group and MCAO/R group were given the injection of the same dose of normal saline. At 2 h and 1

21 and 28 d after modeling

modified neurological severity score (mNSS) was employed to evaluate neurologic deficit

and triphenyl tetrazolium chloride (TTC) staining was performed to measure cerebral infarct volume. The expression of TNF-α (M1 molecule) and CD206 (M2 molecule) in the MG in the brain tissue at 28 d at both mRNA and protein levels were detected.

Results

BMMSCs and BMMSCs-Exo were successfully obtained and identified. MiR-124-1 was significantly expressed in Exo/124-1 rats. All rats (except those from the sham operation group) had neurological deficit after modeling. From 7 d to 28 d after modeling

the mNSS was significantly lower in the Exo/124-1 group than the MCAO/R group (

0.05) and the Exo group and the Exo-Elv group (

0.01). At 28 d after modeling

the infarct volume was notably smaller and the TNF-α expression level was significantly lower in the Exo/124-1 group than the MCAO/R group (

0.01) and the Exo group and Exo-Elv group (

0.01)

and the CD206 level was remarkably higher than the MCAO/R group (

0.01) and the Exo group and Exo-Elv group (

0.01).

Conclusion

BMMSCs-Exo carrying miR-124-1 may regulate the polarization of MG to M2 type

inhibit M1 type-mediated inflammatory response

and thus promote the recovery of neurological function in stroke rats.

关键词

Keywords

references

PAUL S, CANDELARIO-JALIL E. Emerging neuroprotective strategies for the treatment of ischemic stroke: an overview of clinical and preclinical studies[J]. Exp Neurol, 2021, 335: 113518. DOI: 10.1016/j.expneurol.2020.113518.

GVLKE E, GELDERBLOM M, MAGNUS T. Danger signals in stroke and their role on microglia activation after ischemia[J]. Ther Adv Neurol Disord, 2018, 11: 1756286418774254. DOI: 10.1177/1756286418774254.

MA Y Y, WANG J X, WANG Y T, et al . The biphasic function of microglia in ischemic stroke[J ] . Prog Neurobiol, 2017, 157: 247-272. DOI: 10.1016/j.pneurobio.2016.01.005.

KANAZAWA M, NINOMIYA I, HATAKEYAMA M, et al . Microglia and monocytes/macrophages polarization reveal novel therapeutic mechanism against stroke[J ] . Int J Mol Sci, 2017, 18(10): 2135. DOI: 10.3390/ijms18102135.

LI Z W, SONG Y Y, HE T T, et al . M2 microglial small extracellular vesicles reduce glial scar formation via the miR-124/STAT3 pathway after ischemic stroke in mice[J ] . Theranostics, 20 21, 11(3): 1232-1248. DOI: 10.7150/thno.48761..

CHUNG J W, CHANG W H, BANG O Y, et al . Efficacy and safety of intravenous mesenchymal stem cells for ischemic stroke[J ] . Neurology, 2021, 96(7): e1012-e1023. DOI: 10.1212/WNL.0000000000011440.

KRUGER M J, CONRADIE M M, CONRADIE M, et al . ADSC-conditioned media elicit an ex vivo anti-inflammatory macrophage response[J ] . J Mol Endocrinol, 2018, 61(4): 173-184. DOI: 10.1530/jme-18-0078..

HU Y, RAO S S, WANG Z X, et al . Exosomes from human umbilical cord blood accelerate cutaneous wound healing through miR-21-3p-mediated promotion of angiogenesis and fibroblast function[J ] . Theranostics, 2018, 8(1): 169-184. DOI: 10.7150/thno.21234.

NAKAO Y, FUKUDA T, ZHANG Q Z, et al . Exosomes from TNF-α-treated human gingiva-derived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss[J ] . Acta Biomater, 2021, 122: 306-324. DOI: 10.1016/j.actbio.2020.12.046.

郝磊, 金戈, 杨涌涛, 等. 骨髓间充质干细胞外泌体介导miR-124-1对小胶质细胞M2型极化调控的影响[J]. 上海交通大学学报(医学版), 2022, 42(3): 323-330. DOI: 10.3969/j.issn.1674-8115.2022.03.009.

HAO L, JIN G, YANG Y T, et al . Effect of miR-124-1 mediated by exosomes of bone marrow-derived mesenchymal stem cells on the regulation of transformation of M2 microglia[J ] . J Shanghai Jiaotong Univ Med Sci, 2022, 42(3): 323-330. DOI: 10.3969/j.issn.1674-8115.2022.03.009.

郝磊, 田洪, 牟长河, 等. 沉默CX3CL1基因对骨髓间充质干细胞生长及其趋化效应的影响[J]. 第三军医大学学报, 2017, 39(1): 34-41. DOI: 10.16016/j.1000-5404.201607129.

HAO L, TIAN H, MOU C H, et al . Effect of lentivirus-mediated silencing of CX3CL1 gene on growth and chemotaxis effect in bone marrow derived mesenchymal stem cells[J ] . J Third Mil Med Univ, 2017, 39(1): 34-41. DOI: 10.16016/j.1000-5404.201607129..

LIANG Q J, JIANG M, WANG X H, et al . Pre-existing interleukin 10 in cerebral arteries attenuates subsequent brain injury caused by ischemia/reperfusion[J ] . IUBMB Life, 2015, 67(9): 710-719. DOI: 10.1002/iub.1429.

ZHAO Y M, GAN Y X, XU G W, et al . Exosomes from MSCs overexpressing microRNA-223-3p attenuate cerebral ischemia through inhibiting microglial M1 polarization mediated inflammation[J ] . Life Sci, 2020, 260: 118403. DOI: 10.1016/j.lfs.2020.118403.

LIU L R, LIU J C, BAO J S, et al . Interaction of microglia and astrocytes in the neurovascular unit[J ] . Front Immunol, 2020, 11: 1024. DOI: 10.3389/fimmu.2020.01024.

XIONG X Y, LIU L, YANG Q W. Functions and mechanisms of microglia/macrophages in neuroinflammation and neurogenesis after stroke[J]. Prog Neurobiol, 2016, 142: 23-44. DOI: 10.1016/j.pneurobio.2016.05.001.

WEN Z Z, MAI Z, ZHU X L, et al . Mesenchymal stem cell-derived exosomes ameliorate cardiomyocyte apoptosis in hypoxic conditions through microRNA144 by targeting the PTEN/AKT pathway[J ] . Stem Cell Res Ther, 2020, 11(1): 36. DOI: 10.1186/s13287-020-1563-8.

NASSER M I, MASOOD M, ADLAT S, et al . Mesenchymal stem cell-derived exosome microRNA as therapy for cardiac ischemic injury[J ] . Biomed Pharmacother, 2021, 143: 112118. DOI: 10.1016/j.biopha.2021.112118.

VARGAS J, SPIOTTA A, FARGEN K, et al . Long term experience using the ADAPT technique for the treatment of acute ischemic stroke[J ] . J Neurointerv Surg, 2016, 9: 437-441. DOI: 10.1136/neurintsurg-2015-012211.

TANG J Z, LIN X, ZHONG J Y, et al . MiR-124 regulates the osteogenic differentiation of bone marrow-derived mesenchymal stem cells by targeting Sp7[J ] . Mol Med Rep, 2019, 19(5): 3807-3814. DOI: 10.3892/mmr.2019.10054.

LU Z B, LIU S H, OU J Y, et al . Forsythoside A inhibits adhesion and migration of monocytes to type Ⅱ alveolar epithelial cells in lipopolysaccharide-induced acute lung injury through upregulating miR-124[J ] . Toxicol Appl Pharmacol, 2020, 407: 115252. DOI: 10.1016/j.taap.2020.115252.

PERIYASAMY P, LIAO K, KOOK Y H, et al . Cocaine-mediated downregulation of miR-124 activates microglia by targeting KLF4 and TLR4 signaling[J ] . Mol Neurobiol, 2018, 55(4): 3196-3210. DOI: 10.1007/s12035-017-0584-5.

NASIRISHARGH A, KUMAR P, RAMASUBRAMANIAN L, et al . Exosomal microRNAs from mesenchymal stem/stromal cells: biology and applications in neuroprotection[J ] . World J Stem Cells, 2021, 13(7): 776-794. DOI: 10.4252/wjsc.v13.i7.776.

YANG Y X, YE Y Q, FAN K X, et al . miR-124 reduced neuroinflammation after traumatic brain injury by inhibiting TRAF 6 [J ] . Neuroimmunomodulation, 2023, 30(1): 55-68. DOI: 10.1159/000528502..

HAMZEI TAJ S, KHO W, ASWENDT M, et al . Dynamic modulation of microglia/macrophage polarization by miR-124 after focal cerebral ischemia[J ] . J Neuroimmune Pharmacol, 2016, 11(4): 733-748. DOI: 10.1007/s11481-016-9700-y.

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过表达miR-124-1基因的骨髓间充质干细胞外泌体调控小胶质细胞M2型极化对脑卒中的影响

Effect of exosomes derived from mesenchymal stem cells with miR-124-1 overexpression on rat model of stroke by regulating M2 polarization of microglia

DOI：10.16016/j.2097-0927.202305003

摘要

Abstract

关键词

Keywords

references