基础医学与临床 ›› 2022, Vol. 42 ›› Issue (1): 94-99.

• 研究论文 • 上一篇    下一篇

特异性坏死性凋亡抑制剂-1(Nec-1)减轻脊髓损伤模型大鼠胶质瘢痕的形成

陈胜,王春明,严雪飞,毛渊青   

  1. 南京医科大学附属江苏盛泽医院
  • 收稿日期:2020-11-09 修回日期:2021-05-07 出版日期:2022-01-05 发布日期:2022-01-05
  • 通讯作者: 陈胜 E-mail:c2jjqc8@163.com
  • 基金资助:
    南京医科大学康达医药卫生发展研究院科学研究计划项目

Specific necroptosis inhibitor-1 (Nec-1)attenuates glial scar formation in rat models with spinal cord injury

  • Received:2020-11-09 Revised:2021-05-07 Online:2022-01-05 Published:2022-01-05

摘要: 目的:观察特异性坏死性凋亡抑制剂-1(Nec-1)对脊髓损伤大鼠胶质瘢痕形成的影响并探讨其机制。方法:将大鼠随机分为假手术组、模型组、RIP1 K抑制剂Nec-1组。脊髓损伤模型组构建成功后,分别在大鼠侧脑室注射0.9%氯化钠溶液、10μmol/L Nec-1各10μL,假手术组鼠只打开椎板,不打击脊髓。在术后,1、3、7、14d通过BBB检测法测定大鼠后肢运动功能,免疫荧光检测胶质瘢痕形成情况,通过Western blot检测CathepsinB、Caspase-3、GFAP、vimentin表达。结果:术后14d,与假手术组相比,模型组、Nec-1组BBB评分值均显著降低(P<0.05),与模型组相比,Nec-1组BBB评分值显著升高(P<0.05)。术后第14d,假手术组未见NF-200荧光,模型组、Nec-1组均可见NF-200荧光,与模型组相比,Nec-1组NF-200荧光标记强度显著降低(P<0.05)。术后第7d、14d,与假手术组相比,模型组、Nec-1组CathepsinB、Caspase-3、GFAP、vimentin蛋白水平显著升高(P<0.05),与模型组相比,Nec-1组CathepsinB、Caspase-3、GFAP、vimentin蛋白水平显著降低(P<0.05)。结论:Nec-1可能通过调控CathepsinB-Caspase通路,降低GFAP、vimentin蛋白表达,减轻大鼠脊髓损伤后胶质瘢痕形成,促进神经功能恢复。

Abstract: Objective: To observe the effect of necroptosis inhibitor-1 (Nec-1) on glial scar formation in rats with spinal cord injury and explore its mechanism. Methods: The rats were randomly divided into sham operated group, model group and RIP1 K inhibitor Nec-1 group. After the spinal cord injury model group was successfully constructed, 0.9% sodium chloride solution and 10 μmol/L Nec-1 were injected into the lateral ventricles of the rats, respectively. In the sham operation group, rats only were opened the lamina and did not hit the spinal cord. At 1st,3rd,7th,14th day after operation, the motor function of the hind limbs was measured by BBB, the formation of glial scar was detected by immunofluorescence, and the expressions of CathepsinB, Caspase-3, GFAP and vimentin were detected by Western blot. Results: On the 14th day after operation, compared with the sham operation group, the BBB scores of the model group and Nec-1 group were significantly lower (P < 0.05), compared with the model group, the BBB score of Nec-1 group was significantly higher (P < 0.05). On the 14th day after operation, NF-200 fluorescence was not found in the sham operation group, but in the model group and Nec-1 group, compared with the model group, the intensity of NF-200 fluorescence labeling in Nec-1 group decreased significantly (P < 0.05). On the 7th and 14th day after operation, compared with the sham operation group, the protein levels of CathepsinB, Caspase-3, GFAP and vimentin in the model group and Nec-1 group were significantly higher (P < 0.05), compared with the model group, the protein levels of CathepsinB, Caspase-3, GFAP and vimentin in Nec-1 group decreased significantly (P < 0.05). Conclusions: Nec-1 may regulate CathepsinB-Caspase pathway, reduce the expressions of GFAP and vimentin proteins, reduce the formation of glial scar after spinal cord injury, and promote the recovery of neural function.