人脐带间充质干细胞来源外泌体对脑瘫小鼠神经功能损伤的保护作用

doi:10.16352/j.issn.1001-6325.2023.08.1201

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (8): 1201-1207.doi: 10.16352/j.issn.1001-6325.2023.08.1201

人脐带间充质干细胞来源外泌体对脑瘫小鼠神经功能损伤的保护作用

陈星星, 赛依帕^*, 胡筱霞, 王三萍, 罗璇, 刘璟

甘肃省人民医院甘肃中医药大学第一临床医学院儿科,甘肃兰州 730000

收稿日期:2022-08-01 修回日期:2022-12-01 出版日期:2023-08-05 发布日期:2023-07-26
通讯作者: ^*suade10@sohu.com
基金资助:
甘肃省自然科学基金(22JR5RA689);甘肃省青年科技基金计划(22JR5RA701);甘肃省人民医院院内科研基金(20GSSY1-13);甘肃省外科肿瘤分子诊断与精准治疗重点实验室项目(GSSZDSYS2020-10)

Neuroprotective effects of exosomes derived from human umbilical cord mesenchymal stem cells in cerebral palsy mice

CHEN Xingxing, SAI Yipa^*, HU Xiaoxia, WANG Sanping, LUO Xuan, LIU Jing

Department of Pediatrics, People's Hospital of Gansu Province, the First Clinical Medical College of Gansu University of Chinese Medicine, Lanzhou 730000, China

Received:2022-08-01 Revised:2022-12-01 Online:2023-08-05 Published:2023-07-26
Contact: ^*suade10@sohu.com

摘要/Abstract

摘要： 目的探讨人脐带间充质干细胞(HUCMSCs)外泌体对脑性瘫痪(简称脑瘫,CP)小鼠神经功能损伤的保护作用及其机制。方法 HUCMSCs培养、鉴定后提取其外泌体。透射电镜(TEM)、纳米颗粒追踪分析仪(NTA)和Western blot鉴定外泌体;将小鼠随机分为假手术组(sham组,n＝6)、模型组[CP组,n=8,单侧颈总动脉结扎联合脂多糖(LPS)感染后缺氧诱导的方法建立CP模型]、模型-外泌体组(CP-Exo组,n=8,尾静脉注射125 μg外泌体)。Longa评分、网格试验和爬杆试验评估小鼠的神经运动功能、四肢抓握能力和平衡能力;苏木精-伊红(HE)染色光镜下观察脑组织病理损伤,透射电镜观察神经元突触结构,甲苯胺蓝染色检测神经元存活数目,Western blot检测突触相关蛋白突触后致密物95(PSD-95)、突触小泡蛋白(SYP)的表达。结果 HUCMSCs外泌体为形态完整、球形、双层膜囊泡结构,平均粒径123 nm,外泌体TSG101蛋白表达阳性;Longa评分CP组小鼠得分较sham组显著增加(P＜0.01);CP-Exo组评分显著低于CP组(P＜0.05);与CP组相比,CP-Exo组未见斑片状坏死且白质区神经纤维脱髓鞘发生率明显减低(P＜0.01);脑组织结构中sham组突触结构相对正常,另外两组样本均存在不同程度的病变,其病变的大致趋势为:CP组＞CP-Exo组＞sham组;CP组脑组织海马区锥体细胞尼氏小体数量显著低于sham组(P＜0.01),CP-Exo组脑组织海马区锥体细胞尼氏小体数量显著高于CP组(P＜0.01);而CP组PSD-95和SYP表达水平显著低于sham组(P＜0.01),CP-Exo组PSD-95和SYP表达显著高于CP组(P＜0.01)。结论 HUCMSCs外泌体可促进脑瘫小鼠神经功能的恢复,可能通过增加PSD-95和SYP蛋白的表达减轻神经突触的损伤,保护神经元,从而达到改善脑瘫小鼠运动功能的目的。

关键词: 脑性瘫痪, 外泌体, 脐带间充质干细胞

Abstract: Objective To investigate the neuroprotective effects of exosomes derived from human umbilical cord mesenchymal stem cells(HUCMSCs) in cerebral palsy (CP) mice. Methods The exosomes from HUCMSCs were verified by transmission electron microscopy (TEM), nanoparticle tracking analyzer(NTA) and Western blot. The animals were randomly divided as sham operation group (sham group, n=8), model group[CP group, n=8, hypoxic induction after unilateral common carotid artery ligation combined with lipopolysaccharide(LPS) infection] and model exosome group (CP-Exo group, n=8, 125 μg exosomes was injected intravenously). Neuromotor function, the ability of grasping and balance were evaluated by Longa score, grid test and pole climbing test. The pathological lesion in brain tissue was observed with hematoxylin-eosin (HE) staining, the synaptic structure of neurons was observed by TEM, the survival of neurons was detected by toluidine blue staining, and the expression levels of PSD-95 and synaptophysin (SYP) were detected by Western blot. Results The exosomes isolated were spherical and double-membrane vesicles with an average particle size of 123nm, which expressed TSG101 protein. In animal behavior tests, Longa score in CP group was significantly higher than sham group(P＜0.01) and CP-Exo group (P＜ 0.05). There was no plaque necrosis found in exosome group and the incidence of nerve fiber demyelination in the white matter area was significantly reduced than in the CP group (P＜0.01). The synaptic structure of the sham group was relatively normal, while the other two groups showed varying degrees of pathological changes. The general trend of pathological changes was CP group＞CP-Exo group＞ sham group. The number of Nissl bodies in hippocampal pyramidal cells in CP group was significantly less than that in sham group(P＜0.01), however, which was also less than that in CP-Exo group (P＜ 0.01). The expression of PSD-95 and SYP in CP group was significantly lower than that in sham group (P＜0.01), and the levels PSD-95 and SYP in CP-Exo group was significantly higher than those in CP group(P＜0.01). Conclusions HUCMSCs exosomes can promote the recovery of neural function of cerebral palsy mice, reduce synaptic damage and protect neuron cells by increasing the expression of PSD-95 and SYP.

Key words: cerebral palsy, exosomes, human umbilical cord mesenchymal stem cells(HUCMSCs)

中图分类号:

R742
R364

陈星星, 赛依帕, 胡筱霞, 王三萍, 罗璇, 刘璟. 人脐带间充质干细胞来源外泌体对脑瘫小鼠神经功能损伤的保护作用[J]. 基础医学与临床, 2023, 43(8): 1201-1207.

CHEN Xingxing, SAI Yipa, HU Xiaoxia, WANG Sanping, LUO Xuan, LIU Jing. Neuroprotective effects of exosomes derived from human umbilical cord mesenchymal stem cells in cerebral palsy mice[J]. Basic & Clinical Medicine, 2023, 43(8): 1201-1207.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: http://journal11.magtechjournal.com/Jwk_jcyxylc/CN/10.16352/j.issn.1001-6325.2023.08.1201

http://journal11.magtechjournal.com/Jwk_jcyxylc/CN/Y2023/V43/I8/1201

参考文献

[1]中国康复医学会儿童康复专业委员会,中国残疾人康复协会小儿脑性瘫痪康复专业委员会,中国医师协会康复医师分会儿童康复专业委员会,等. 中国脑性瘫痪康复指南(2022)第一章:概论[J]. 中华实用儿科临床杂志,2022,37:887-892.
[2]李晓捷,邱洪斌,姜志梅, 等.中国十二省市小儿脑性瘫痪流行病学特征[J]. 中华实用儿科临床杂志, 2018, 33: 378-383.
[3]Morgan C, Fahey M, Roy B, et al. Diagnosing cerebral palsy in full-term infants[J].J Paediatr Child Health, 2018, 54: 1159-1164.
[4]Dong L, Peng Z, Yao X, et al. Exosomes derived from mir-133b-modified mesenchymal stem cells promote recovery after spinal cord injury[J]. Front Neurosci, 2018,12: 845. doi:10.3389/fnins.2018.00845.
[5]Feather-Schussler DN, Ferguson TSA battery of motor tests in a neonatal mouse model of cerebral palsy[J]. J Vis Exp, 2016, 117: e53569. doi:10.3791/53569.
[6]Niewiadomski W, Palasz E, Skupinska M, et al. TracMouse: a computer aided movement analysis script for the mouse inverted horizontal grid test[J]. Sci Rep, 2016, 6, 39331. doi:10.1038/srep39331.
[7]Gao Y, Tang H, Nie K, et al. Hippocampal damage and white matter lesions contribute to cognitive impairment in MPTP-lesioned mice with chronic cerebral hypoperfusion[J].Behav Brain Res, 2019,368:111885. doi:10.1016/j.bbr.2019.03.054.
[8]Tsiapalis D, O'Driscoll L. Mesenchymal stem cell derived extracellular vesicles for tissue engineering and regenerative medicine applications[J].Cells, 2020, 9:991. doi:10.3390/cells9040991.
[9]高振橙,刘欣.间充质干细胞外泌体在神经系统疾病修复过程中的作用与应用[J].中国组织工程研究,2020,24:3048-3054.
[10]Volpe JJ. Confusions in nomenclature: “periventricular leukomalacia” and “white matter injury”-identical, distinct, or overlapping?[J].Pediatr Neurol, 2017,73:3-6. doi:10.1016/j.pediatrneurol.2017.05.013.
[11]Filley CM, Fields RD. White matter and cognition: making the connection[J].J Neurophysiol, 2016, 116:2093-2104.doi:10.1152/jn.00221.2016.
[12]Thiel G. Synapsin I, synapsin II, and synaptophysin: marker proteins of synaptic vesicles[J]. Brain Pathol, 1993, 3:87-95. doi:10.1111/j.1750-3639.1993.tb00729.x.
[13]Ulfig N, Setzer M, Neudörfer F, et al. Distribution of SNAP-25 in transient neuronal circuitries of the develop-ing human forebrain. Neuroreport, 2000,11:1259-1263.doi:10.1097/00001756-200004270-00023.

[1]	闫霖, 陆珏秀, 罗颖, 刘先霞. BM-MSCs来源外泌体介导铁死亡减轻大鼠心肌细胞系H9c2缺氧/复氧损伤[J]. 基础医学与临床, 2023, 43(5): 771-776.
[2]	崔诗雨, 张海峰. 外泌体在新型冠状病毒感染和传播中作用的研究进展[J]. 基础医学与临床, 2023, 43(4): 680-684.
[3]	姚刘旭, 李玉红, 蒋宗明. 外泌体lncRNA在结直肠癌液体活检中的研究进展[J]. 基础医学与临床, 2023, 43(2): 317-321.
[4]	姜俣, 钱海燕. 间充质干细胞治疗心肌梗死的研究进展[J]. 基础医学与临床, 2023, 43(1): 21-29.
[5]	商利鹏, 刘国祥, 丁晓燕, 李冰, 赵春华, 李霄霞. 间充质干细胞在衰老相关疾病应用中的研究进展[J]. 基础医学与临床, 2023, 43(1): 38-45.
[6]	陈莉莉, 孙永红, 雷晓燕, 王建军, 陈星星, 鲍柏辛. 间充质干细胞外泌体治疗肺部疾病的研究进展[J]. 基础医学与临床, 2023, 43(1): 192-195.
[7]	吴文哲, 孙建伟, 付长秀, 黄李晨露, 郑永钦, 贺军栋. 脐带间充质干细胞修复皮肤创伤的研究进展[J]. 基础医学与临床, 2022, 42(11): 1771-1775.
[8]	李英杰, 杨庭楷, 纪志刚, 文进. 外泌体在前列腺癌诊疗中的研究进展[J]. 基础医学与临床, 2022, 42(1): 163-167.
[9]	杨旭东, 于红亮, 李静, 赵春华. 巨噬细胞J774A.1外泌体促进结直肠癌细胞系MC38和CT26的增殖和迁移[J]. 基础医学与临床, 2021, 41(6): 799-804.
[10]	赵晓龙, 代晓燕, 毛成毅, 肖华亮, 王东, 戴楠. 非小细胞肺癌患者血浆外泌体APE1的表达与铂类化疗敏感性相关[J]. 基础医学与临床, 2021, 41(2): 165-170.
[11]	朱萌, 张宁, 陶伟. 人胃癌细胞系AGS外泌体miR-106a对腹膜间皮细胞系HMrSV5表型的影响[J]. 基础医学与临床, 2021, 41(11): 1564-1569.
[12]	董保龙, 韩彩文, 史明, 郑文学, 杨晓军. 波形蛋白在肝细胞癌中的研究进展[J]. 基础医学与临床, 2020, 40(2): 238-242.
[13]	杨玥, 卞涛. 不同细胞来源的外泌体在COPD中的研究进展[J]. 基础医学与临床, 2020, 40(12): 1721-1726.
[14]	刁华琼, 李小黎, 林亮吟, 刘海鹏, 陈弘婧, 丁海月, 魏丹. 外泌体microRNAs与阿尔茨海默病关系的研究进展[J]. 基础医学与临床, 2020, 40(11): 1542-1545.
[15]	顾国民布力布?吉力斯汉卢素琼王秀丽刘春玲. 肿瘤微环境中Lewis肺腺癌细胞调控巨噬细胞表型转换[J]. 基础医学与临床, 2019, 39(9): 1252-1258.

人脐带间充质干细胞来源外泌体对脑瘫小鼠神经功能损伤的保护作用

Neuroprotective effects of exosomes derived from human umbilical cord mesenchymal stem cells in cerebral palsy mice

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价