摘要
目的观察mi R-429-5p在脊髓损伤后的表达变化及其对脊柱神经元细胞的凋亡的影响,并探讨mi R-429-5p在脊髓损伤中的作用机制。方法(1)培养大鼠背根神经节细胞,通过qPCR检测mi R-429-5p在大鼠背根神经节细胞受损后的表达变化,并通过流式细胞仪检测mi R-429-5p对受损背根神经节细胞的凋亡的影响,同时通过Western blot检测Caspase3的蛋白表达;(2)通过TargetScan基因库预测mi R-429-5p的靶基因为Bcl2,通过双荧光素酶实验验证其靶向关系,并通过Western blot检测mi R-429-5p对Bcl2表达的调控;(3)利用SD大鼠建立脊髓损伤模型,正常组10只,脊髓损伤模型组20只,mi R-429-5p Inhibitor组20只。进行BBB评分检测各组后肢运动的恢复情况,通过qPCR检测mi R-429-5p以及Bcl2的表达变化,通过Western blot检测凋亡相关基因Caspase3的变化,通过嗜银染色和HE染色观察神经纤维的再生。结果(1)miR-429-5p参与脊髓损伤后的再生过程,mi R-429-5p的表达下调(P<0.05);(2)miR-429-5p参与调控受损背根神经节细胞的凋亡,mi R-429-5p的表达下调(P<0.05)。结论 mi R-429-5p通过靶向Bcl2参与调控脊髓损伤过程中神经元细胞的凋亡,从而影响脊髓神经细胞功能的再生,为继发性脊髓损伤的治疗提供了新的靶点。
Objective To investigate the expression alternation of miR-429-5 p after the spinal cord injury and the effect of the miR-429-5 p on apoptosis of spinal neurons after the spinal cord injury, and to explore the mechanism of miR429-5 p in the spinal cord injury. Methods(1) The expression of miR-429-5 p in the injured DRG cells was detected by qPCR, the effect of miR-429-5 p on the apoptosis of DRG cells was detected by flow cytometry, and the expression of Caspase3 was detected by Western blot.(2) The target relation between miR-429-5 p and Bcl2 was predicted by TargetScan, and then verified by double luciferase assay, and the expression of Bcl2 was detected by Western blot.(3)SD rats were used to establish the spinal cord injury model,including 10 rats in the normal group, 20 rats in the spinal cord injury model group and 20 rats in the miR-429-5 p the In hibitor group. The recovery of hindlimb movement was measured by BBB score, the expression of miR-429-5 p and Bcl2 were detected by q PCR. The protein expression of caspase3 was detected by Western blot, and the regeneration of nerve fibers was observed by silver staining and HE staining. Results(1)miR-429-5 p was involved in the regeneration process after the spinal cord injury, and the down regulation of miR-429-5 p could promote the recovery of hindlimb function.(2)miR-429-5 p was involved in the regulation of apoptosis of DRG cells. When the expression of miR-429-5 p was down regulated, the apoptosis of DRG cells was significantly alleviated. Conclusion miR-429-5 p was involved in the regulation of neuron apoptosis in the regeneration process of spinal cord injury by targeting Bcl2, so as to involve the regeneration of neural cell function in the spinal cord and provided a new target for the treatment of spinal cord injury.
作者
陶裕川
易勇
周世军
王翰
TAO Yu-chuan;YI Yong;ZHOU Shi-jun;WANG Han(Dept.of Neurosurgery,Yibin Second People’s Hospital,Yibin Sichuan 644000,China)
出处
《昆明医科大学学报》
CAS
2020年第10期21-27,共7页
Journal of Kunming Medical University
基金
宜宾市重点科技基金资助项目(2017ZSF007-5)。
作者简介
陶裕川(1976-)男,四川宜宾人,医学硕士,副主任医师,主要从事神经脊柱外科临床工作;通信作者:王翰,E-mail:284653117@qq.com。
