中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (2): 396-403.doi: 10.4103/1673-5374.346463

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

敲低多嘧啶束结合蛋白可促进脊髓损伤后运动功能的恢复

  

  • 出版日期:2023-02-15 发布日期:2022-08-09
  • 基金资助:
    国家自然科学基金项目（82101455，31872773，82001168）；江苏省重点研发计划项目（社会发展）（BE2020667）； 江苏省“333工程高层次人才”基金项目（BRA2020076）；南通市科技项目（JC2020028）；江苏省高等学校自然科学研究项目（19KJB310012）；江苏省高等学校重点学术项目发展项目（PAPD）

Knockdown of polypyrimidine tract binding protein facilitates motor function recovery after spinal cord injury

Ri-Yun Yang1, Rui Chai2, Jing-Ying Pan1, Jing-Yin Bao3, Pan-Hui Xia3, Yan-Kai Wang3, Ying Chen1, Yi Li1, Jian Wu1, Gang Chen2, 3, 4, *   

  1. 1Department of Histology and Embryology, Medical School of Nantong University, Nantong, Jiangsu Province, China; 2Key Laboratory of Neuroregeneration of Jiangsu Province and the Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China; 3Center for Basic Medical Research, Medical School of Nantong University, Nantong, Jiangsu Province, China; 4Department of Anesthesiology, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China
  • Online:2023-02-15 Published:2022-08-09
  • Contact: Gang Chen, MD, PhD, chengang6626@ntu.edu.cn.
  • Supported by:
    The study was supported by the National Natural Science Foundation of China, Nos. 82101455 (to RYY), 31872773 (to GC), 82001168 (to JYP); the Key Research and Development Program (Social Development) of Jiangsu Province, No. BE2020667 (to GC); the Foundation of Jiangsu Province “333 Project High-level Talents”, No. BRA2020076 (to GC); the Nantong Civic Science and Technology Project of China, No. JC2020028 (to RYY); the Natural Science Research of Jiangsu Higher Education Institutions of China, No. 19KJB310012 (to RYY) and Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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摘要：

脊髓损伤后，病灶核心处可形成以纤维细胞和小胶质细胞介导的纤维化瘢痕，且由于星形胶质细胞的活化和增殖，在纤维化瘢痕周围可形成胶质 瘢痕。同时，损伤部位存在大量神经元缺失。调节胶质瘢痕的密度和补充损伤区域的运动神经元对于脊髓损伤修复至关重要。多嘧啶束结合蛋白是一种RNA结合蛋白，可在神经发生中起重要作用。此次实验首先在体外利用病毒搭载的短发夹RNA和反义寡核苷酸来敲低多嘧啶束结合蛋白的表达，成功地将小鼠脊髓反应性星形胶质细胞重编程为运动神经元样细胞。而后在压迫性脊髓损伤小鼠模型中，腺相关病毒短发夹RNA敲低多嘧啶束结合蛋白可增加损伤区域附近的运动神经元样细胞数量，还促进小鼠运动功能的恢复；而反义寡核苷酸敲低多嘧啶束结合蛋白的表达，不仅可以改善脊髓损伤小鼠的运动功能恢复，补充损伤区域周围的运动神经元样细胞，而且在不破坏其整体结构的情况下适度降低胶质瘢痕的密度。这一结果预示着多嘧啶束结合蛋白敲低可能是一种有前途的治疗脊髓损伤的策略。

https://orcid.org/0000-0003-3669-5687 (Gang Chen)

关键词: 脊髓修复, 星形胶质细胞, 胶质瘢痕, 多嘧啶束结合蛋白, 神经发生, 神经元样细胞, 运动神经元样细胞, 运动功能, 短发夹RNA, 反义寡核苷酸

Abstract: After spinal cord injury (SCI), a fibroblast- and microglia-mediated fibrotic scar is formed in the lesion core, and a glial scar is formed around the fibrotic scar as a result of the activation and proliferation of astrocytes. Simultaneously, a large number of neurons are lost in the injured area. Regulating the dense glial scar and replenishing neurons in the injured area are essential for SCI repair. Polypyrimidine tract binding protein (PTB), known as an RNA-binding protein, plays a key role in neurogenesis. Here, we utilized short hairpin RNAs (shRNAs) and antisense oligonucleotides (ASOs) to knock down PTB expression. We found that reactive spinal astrocytes from mice were directly reprogrammed into motoneuron-like cells by PTB downregulation in vitro. In a mouse model of compression-induced SCI, adeno-associated viral shRNA-mediated PTB knockdown replenished motoneuron-like cells around the injured area. Basso Mouse Scale scores and forced swim, inclined plate, cold allodynia, and hot plate tests showed that PTB knockdown promoted motor function recovery in mice but did not improve sensory perception after SCI. Furthermore, ASO-mediated PTB knockdown improved motor function restoration by not only replenishing motoneuron-like cells around the injured area but also by modestly reducing the density of the glial scar without disrupting its overall structure. Together, these findings suggest that PTB knockdown may be a promising therapeutic strategy to promote motor function recovery during spinal cord repair.

Key words: antisense oligonucleotides, astrocytes, glial scar, motoneuron-like cells, motor function, neurogenesis, neuron-like cells, polypyrimidine tract binding protein, short hairpin RNAs, spinal cord repair