中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (30): 4793-4799.doi: 10.12307/2022.758

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

脐带间充质干细胞促进蜕膜血管内皮生长因子表达改善自然流产小鼠妊娠结局

赵鑫鑫,杨晓清,张玉泉   

  1. 南通大学附属医院妇产科,江苏省南通市   226001
  • 收稿日期:2021-07-26 接受日期:2021-09-13 出版日期:2022-10-28 发布日期:2022-03-29
  • 通讯作者: 张玉泉,博士生导师,主任医师,教授,南通大学附属医院妇产科,江苏省南通市 226001
  • 作者简介:赵鑫鑫,女,1989年生,江苏省南通市人,汉族,2020年南通大学毕业,硕士,医师,主要从事早期自然流产方向的研究。
  • 基金资助:
    江苏省重点研发计划(社会发展) 面上项目(BE2018672),项目负责人:张玉泉

Umbilical cord mesenchymal stem cells improve pregnancy outcomes of spontaneous abortion mouse models by promoting vascular endothelial growth factor expression in decidua

Zhao Xinxin, Yang Xiaoqing, Zhang Yuquan   

  1. Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China
  • Received:2021-07-26 Accepted:2021-09-13 Online:2022-10-28 Published:2022-03-29
  • Contact: Zhang Yuquan, Doctoral supervisor, Chief physician, Professor, Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China
  • About author:Zhao Xinxin, Master, Physician, Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China
  • Supported by:
    Social Development Foundation of Jiangsu Province, China, No. BE2018672 (to ZYQ)

摘要:

文题释义:
蜕膜:位于胚胎着床的母胎交界面,其异质性较高。蜕膜有着较为复杂的细胞构成,单细胞测序技术揭示人蜕膜中的细胞有蜕膜基质细胞、蜕膜成纤维细胞、平滑肌细胞、子宫内膜上皮细胞、自然杀伤细胞、抗原呈递细胞、T细胞、淋巴管内皮细胞和血管内皮细胞。蜕膜的细胞因子网络在正常妊娠时保持着动态平衡,而该平衡破坏是引起各种不良妊娠结局如自然流产甚至复发性流产等的重要原因。
血管内皮生长因子:是一种重要的促血管生成因子,母胎界面的蜕膜基质细胞、蜕膜免疫细胞及滋养细胞均可分泌血管内皮生长因子,其在胚胎植入过程中发挥重要作用,血管内皮生长因子通过与血管内皮细胞表面血管内皮生长因子受体2结合,促进血管生成,调节血管通透性;此外,血管内皮生长因子可促进蜕膜中巨噬细胞的募集,同时促进M1型巨噬细胞向M2型巨噬细胞转化。有研究提出母胎界面血管内皮生长因子表达减少与自然流产相关。

背景:早期自然流产与母胎界面血管形成障碍或异常有关,人脐带华通氏胶间充质干细胞具有向损伤部位迁移及促血管生成的作用,目前人脐带华通氏胶间充质干细胞作用于早期自然流产蜕膜基质细胞的研究尚未有报道。
目的:探讨人脐带华通氏胶间充质干细胞对早期自然流产蜕膜基质细胞增殖、凋亡及血管内皮生长因子表达的影响,以及对早期自然流产小鼠妊娠结局的影响。
方法:①分离培养早期自然流产和人工流产蜕膜组织,实时荧光定量 PCR和Western blot检测蜕膜组织中血管内皮生长因子的表达;②第3代人脐带华通氏胶间充质干细胞和原代自然流产蜕膜基质细胞共培养72 h,EdU、Annexin V-FITC分别检测蜕膜基质细胞增殖和凋亡情况,实时荧光定量 PCR和Western blot检测蜕膜基质细胞中血管内皮生长因子的表达;③复制CBA/J×DBA/2自然流产小鼠模型,孕1,3,5 d尾静脉移植人脐带华通氏胶间充质干细胞悬液0.1 mL(细胞浓度为1×1010 L-1),孕14 d计算胚胎吸收率,ELISA检测外周血中血管内皮生长因子水平;实时荧光定量 PCR和Western blot检测蜕膜组织中血管内皮生长因子的表达。
结果与结论:①与正常蜕膜组织相比,早期自然流产患者蜕膜中血管内皮生长因子表达显著降低;②人脐带华通氏胶间充质干细胞促进早期自然流产蜕膜基质细胞中血管内皮生长因子的表达,促进细胞增殖、抑制细胞凋亡;③人脐带华通氏胶间充质干细胞促进自然流产小鼠蜕膜组织中血管内皮生长因子表达,进而减少胚胎吸收率;④结果表明,合适剂量的人脐带华通氏胶间充质干细胞可以促进蜕膜中血管内皮生长因子表达,改善早期自然流产妊娠结局。
缩略语:脐带华通氏胶间充质干细胞:Wharton’s jelly-derived mesenchymal stem cells,WJ-MSCs

https://orcid.org/0000-0003-2104-1111 (赵鑫鑫) 

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: 自然流产, 血管内皮生长因子, 蜕膜基质细胞, 人脐带华通氏胶间充质干细胞, 小鼠流产模型

Abstract: BACKGROUND: Early spontaneous abortion is related to the disorder or abnormality of blood vessel formation at the maternal-fetal interface. Human umbilical cord Wharton’s jelly-mesenchymal stem cells can migrate to the injured site and promote angiogenesis. However, the effect of human Wharton’s jelly- mesenchymal stem cells on decidual stromal cells has not been studied yet.
OBJECTIVE: To investigate the effect of human Wharton’s jelly-mesenchymal stem cells on proliferation and apoptosis of decidual stromal cells from early spontaneous abortion women and vascular endothelial growth factor expression, and to explore its effect on the pregnancy outcome of early spontaneous abortion mice. 
METHODS: (1) Early spontaneous abortion and artificial abortion decidual tissues were separated and cultured. Real-time fluorescent quantitative PCR and western blot assay were utilized to detect the expression of vascular endothelial growth factor in decidua tissue. (2) The third passage of human umbilical cord Wharton’s jelly-mesenchymal stem cells and primary spontaneous abortion decidual stromal cells were co-cultured for 72 hours. EdU and Annexin V-FITC were applied to detect the proliferation and apoptosis of decidual stromal cells, respectively. Real-time fluorescent quantitative PCR and western blot assay were used to detect the expression of vascular endothelial growth factor in decidual stromal cells. (3) After reproducing the CBA/J×DBA/2 spontaneous abortion mouse model, human umbilical cord Wharton’s jelly-mesenchymal stem cell suspension 0.1 mL (cell concentration: 1×1010 L-1) was transplanted into the tail vein at 1, 3, and 5 days after pregnancy. The embryo absorption rate was calculated at 14 days of pregnancy. ELISA was used to detect the level of vascular endothelial growth factor in peripheral blood. Real-time fluorescent quantitative PCR and western blot assay were employed to detect the expression of vascular endothelial growth factor in decidua tissue.
RESULTS AND CONCLUSION: (1) Compared with normal decidua tissue, expression of vascular endothelial growth factor was significantly downregulated in the decidua during human early spontaneous abortion. (2) Human Wharton’s jelly-mesenchymal stem cells increased vascular endothelial growth factor expression, promoted proliferation, and inhibited apoptosis in human abortion decidual stromal cells. (3) Human Wharton’s jelly-mesenchymal stem cells promoted the expression of vascular endothelial growth factor in the decidua of spontaneous abortion mice and reduced embryo absorption of the abortion prone mouse model. (4) In summary, a certain amount of Human Wharton’s jelly-mesenchymal stem cells transplanted can increase vascular endothelial growth factor levels in decidua and improve the outcome of the abortion prone mouse model. 

Key words: spontaneous abortion, vascular endothelial growth factor, decidual stromal cells, human Wharton’s jelly-mesenchymal stem cells, abortion prone mouse model

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