中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (9): 2029-2036.doi: 10.4103/1673-5374.367831

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

人牙髓干细胞移植治疗缺血性脑卒中:miR-34a是有价值的候选基因

  

  • 出版日期:2023-09-15 发布日期:2023-03-06
  • 基金资助:
    国自然(81971870;82172173)

Genetic modification of miR-34a enhances efficacy of transplanted human dental pulp stem cells after ischemic stroke

Jianfeng Wang1, #, Peibang He1, #, Qi Tian1, Yu Luo2, Yan He3, Chengli Liu1, Pian Gong1, Yujia Guo1, Qingsong Ye2, *, Mingchang Li1, *   

  1. 1Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China; 2Center of Regenerative Medicine, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China; 3Institute of Regenerative and Translational Medicine, Tianyou Hospital, Wuhan University of Science and Technology, Wuhan, Hubei Province, China
  • Online:2023-09-15 Published:2023-03-06
  • Contact: Qingsong Ye, PhD, qingsongye@whu.edu.cn; Mingchang Li, MD, mingcli@whu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, Nos. 81971870 and 82172173 (both to ML).

摘要:

人牙髓干细胞移植能促进缺血性脑卒中后的恢复;然而,由于移植的细胞存活率低,治疗效果受到限制。体外实验在人牙髓干细胞中使用了氧糖剥夺/复氧来模拟缺血再灌注对细胞造成的损害,发现在此条件下,人牙髓干细胞的miRNA-34a-5p(miR-34a)水平升高了。抑制miR-34a有利于人牙髓干细胞的增殖和抗氧化能力的增强,减少细胞凋亡;WNT1和SIRT1是miR-34a的作用标靶。在miR-34a敲除的细胞系中,miR-34a分别通过靶向WNT1和SIRT1来调控细胞增殖和抗氧化能力。将基因修饰的牙髓干细胞(anti34a-hDPSCs)注入小鼠脑内后发现,anti34a-人牙髓干细胞可显著抑制脑梗死后的细胞凋亡,脑水肿,降低脑梗死体积,改善小鼠的运动功能。通过抑制牙髓干细胞的miR-34a可通过与WNT1和SIRT1翻译的mRNA结合,分别上调WNT1/β-catenin和SIRT1/Nrf2/HO-1信号通路,促进人牙髓干细胞的增殖和抗氧化能力。人牙髓干细胞移植对小鼠缺血性脑卒中产生神经保护作用,特别是miR-34a敲除的人牙髓干细胞。因此,miR-34a是干细胞基因修饰的一个有价值的候选基因,可用于提高移植干细胞治疗缺血性脑卒中的疗效。

https://orcid.org/0000-0002-2868-9247 (Qingsong Ye); https://orcid.org/0000-0003-4019-8886 (Mingchang Li)

Abstract: Human dental pulp stem cells (hDPSCs) promote recovery after ischemic stroke; however, the therapeutic efficacy is limited by the poor survival of transplanted cells. For in vitro experiments in the present study, we used oxygen-glucose deprivation/reoxygenation in hDPSCs to mimic cell damage induced by ischemia/reperfusion. We found that miRNA-34a-5p (miR-34a) was elevated under oxygen-glucose deprivation/reoxygenation conditions in hDPSCs. Inhibition of miR-34a facilitated the proliferation and antioxidant capacity and reduced the apoptosis of hDPSCs. Moreover, dual-luciferase reporter gene assay showed WNT1 and SIRT1 as the targets of miR-34a. In miR-34a knockdown cell lines, WNT1 suppression reduced cell proliferation, and SIRT1 suppression decreased the antioxidant capacity. Together, these results indicated that miR-34a regulates cell proliferation and antioxidant stress via targeting WNT1 and SIRT1, respectively. For in vivo experiments, we injected genetically modified hDPSCs (anti34a-hDPSCs) into the brains of mice. We found that anti34a-hDPSCs significantly inhibited apoptosis, reduced cerebral edema and cerebral infarct volume, and improved motor function in mice. This study provides new insights into the molecular mechanism of the cell proliferation and antioxidant capacity of hDPSCs, and suggests a potential gene that can be targeted to improve the survival rate and efficacy of transplanted hDPSCs in brain after ischemic stroke.

Key words: antioxidant capacity, HO-1, human dental pulp stem cells, ischemic stroke, miR-34a, Nrf2, proliferation, SIRT1, WNT1, β-catenin