人脐带间充质干细胞预防小鼠急性移植物抗宿主病作用及其机制研究

洪涛; 王瑞; 王筱淇; 王蔚浩; 杜雨軒; 何云地; 马超; 安慧慧; 王俊洁; 张云芳; 高强国; 杨世杰; 张曦

doi:10.16016/j.1000-5404.202108059

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人脐带间充质干细胞预防小鼠急性移植物抗宿主病作用及其机制研究

专题报道：造血干细胞移植并发症的基础与临床研究 | 更新时间：2026-01-09

- 人脐带间充质干细胞预防小鼠急性移植物抗宿主病作用及其机制研究
- Effect and mechanism of human umbilical cord mesenchymal stem cells on preventing murine acute graft-versus-host disease
- 第三军医大学学报 2021年43卷第21期页码：2276-2285
- 作者机构：
  
  1.400037 重庆，陆军军医大学(第三军医大学)第二附属医院血液病医学中心，全军血液病中心，创伤、烧伤与复合伤国家重点实验室
  2.563099 贵州遵义，遵义医科大学第二附属医院检验科
  3.400038 重庆，陆军军医大学(第三军医大学)基础医学院细胞生物学教研室
- 作者简介：
  
  [张曦，E-mail: zhangxxi@sina.com ZHANG Xi, E-mail: zhangxxi@sina.com
- 基金信息：
  
  国家重点研究计划重点专项(2017YFA0105502);国家血液系统疾病临床医学研究中心2020年度开放课题重点项目(2020ZKZC02)
- DOI：10.16016/j.1000-5404.202108059
  中图分类号： R392.8;R617;R733.7
- 收稿：2021-08-10，
  
  修回：2021-09-22，
  
  纸质出版：2021-11-15
- 稿件说明：
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洪涛, 王瑞, 王筱淇, 等. 人脐带间充质干细胞预防小鼠急性移植物抗宿主病作用及其机制研究[J]. 第三军医大学学报, 2021,43(21):2276-2285.

Tao HONG, Rui WANG, Xiaoqi WANG, et al. Effect and mechanism of human umbilical cord mesenchymal stem cells on preventing murine acute graft-versus-host disease[J]. Journal of Third Military Medical University, 2021, 43(21): 2276-2285.
洪涛, 王瑞, 王筱淇, 等. 人脐带间充质干细胞预防小鼠急性移植物抗宿主病作用及其机制研究[J]. 第三军医大学学报, 2021,43(21):2276-2285. DOI： 10.16016/j.1000-5404.202108059.

Tao HONG, Rui WANG, Xiaoqi WANG, et al. Effect and mechanism of human umbilical cord mesenchymal stem cells on preventing murine acute graft-versus-host disease[J]. Journal of Third Military Medical University, 2021, 43(21): 2276-2285. DOI： 10.16016/j.1000-5404.202108059.

摘要

目的

探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells

HUCMSCs)对小鼠急性移植物抗宿主病(acute graft-versus-host disease

aGVHD)的预防效果并初步探索其作用机制。

方法

通过对接受全身辐照的雄性BALB/c小鼠移植雄性C57BL/6小鼠的骨髓细胞和脾脏细胞，构建小鼠异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation

allo-HSCT)后aGVHD模型，建模部分实验分组：BM组[尾静脉注射500 μL供者C57BL/6小鼠骨髓细胞(5×10

/只)悬液

]

和aGVHD组[尾静脉注射500 μL骨髓细胞(5×10

/只)和脾脏细胞(5×10

/只)混悬液

]

。观察小鼠生存率、临床评分和体质量变化；流式细胞术检测移植后嵌合度；HE染色观察aGVHD靶器官(肺脏、肝脏、结肠、小肠)的病理学改变。收集P4代HUCMSCs，于移植当天混合供者骨髓细胞和脾脏细胞输注BALB/c小鼠，实验分组：①aGVHD组(处理同建模部分)；②aGVHD+MSCs组[经尾静脉注射500 μL骨髓细胞(5×10

/只)、脾脏细胞(5×10

/只)和HUCMSCs(5×10

/只)混悬液

]

。观察HUCMSCs对aGVHD的预防效果，应用ELISA检测各组小鼠血清IFN-γ、TNF-α浓度；流式细胞术检测CD4

T细胞在靶器官中绝对数计数和凋亡水平。

结果

aGVHD组较BM组生存率降低、临床评分增高、体质量减轻明显(

＜0.05)，靶器官病理损伤更为严重(

＜0.05)；移植后小鼠达完全嵌合。HUCMSCs输注显著改善移植后小鼠生存、降低临床评分和靶器官病理损伤(

＜0.05)，并降低血清炎症因子浓度(

＜0.05)，增加CD4

T细胞在靶器官中的凋亡，CD4

T细胞绝对数计数显著下降(

＜0.05)。

结论

HUCMSCs联合allo-HSCT能有效预防及减轻小鼠aGVHD，其作用机制主要是HUCMSCs介导淋巴细胞凋亡，减少其在靶器官中的浸润和调控炎症因子分泌。

Abstract

Objective

To determine the prophylactic effect of human umbilical cord mesenchymal stem cells (HUCMSCs) on murine acute graft-versus-host disease (aGVHD) and preliminarily explore its mechanism.

Methods

In order to establish a mouse model of aGVHD

the male BALB/c mice were transplanted with bone marrow cells and spleen lymphocytes from male C57BL/6 mice after receiving 7.5 Gy total body irradiation (TBI) on the day of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The mice were divided into BM group and aGVHD group

receiving 500 μL cell suspension containing 5×10

bone marrow cells or 5×10

bone marrow cells and 5×10

spleen cells

respectively

by tail vein. Their survival rate

clinical score and loss of body weight were assessed

chimerism was detected by flow cytometry

and pathological changes of target organs (lung

liver

colon

small intestine) were detected by HE staining. The HUCMSCs at passage 4 were collected and transplanted into aGVHD+MSCs group of BALB/c mice. The mice were then divided into aGVHD group (model group)

aGVHD+MSCs group (5×10

bone marrow cells and 5×10

spleen cells and 5×10

HUCMSCs). Besides

the serum concentration of IFN-γ and TNF-α were detected by ELISA

and the infiltration and apoptosis of lymphocytes were detected by flow cytometry.

Results

Compared with the BM group

the aGVHD group showed a decreased survival rate

higher clinical score

and much obvious body weight loss (

0.05) and severer pathological injuries (

0.05). The mice achieved complete chimerism after transplantation. The aGVHD+MSCs group showed increased survival rate

lower clinical score and slighter pathological injuries (

0.05)

the serum concentrations of IFN-γ and TNF-α were decreased (

0.05)

and the apoptosis of CD4

T cells in target organs was increased while the count was decreased (

0.05).

Conclusion

HUCMSCs infusion effectively prevent the development of murine aGVHD after allo-HSCT through mediating the apoptosis of lymphocytes to decrease their infiltration in target organs and regulating the secretion of inflammatory factors.

关键词

Keywords

references

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