Xiao Wei-Ping, Ding Li-Li-Qiang, Min You-Jiang, Yang Hua-Yuan, Yao Hai-Hua, Sun Jie, Zhou Xuan, Zeng Xue-Bo, Yu Wan
Spinal Department of Orthopedics and Department of Acupuncture, Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi, People's Republic of China.
Department of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Neuropsychiatr Dis Treat. 2019 Dec 13;15:3429-3442. doi: 10.2147/NDT.S216874. eCollection 2019.
To observe the changes of Nogo/NgR and Rho/ROCK signaling pathway-related gene and protein expression in rats with spinal cord injury (SCI) treated with electroacupuncture (EA) and to further investigate the possible mechanism of EA for treating SCI.
Allen's method was used to create the SCI rat model. Sixty-four model rats were further subdivided into four subgroups, namely, the SCI model group (SCI), EA treatment group (EA), blocking agent Y27632 treatment group (Y27632) and EA+blocking agent Y27632 treatment group (EA+Y), according to the treatment received. The rats were subjected to EA and/or blocking agent Y27632 treatment. After 14 days, injured spinal cord tissue was extracted for analysis. The mRNA and protein expression levels were determined by real-time fluorescence quantitative PCR and Western blotting, respectively. Cell apoptosis changes in the spinal cord were evaluated by in situ hybridization. Hindlimb motor function in the rats was evaluated by Basso-Beattie-Bresnahan assessment methods.
Except for RhoA protein expression, compared with the SCI model group, EA, blocking agent Y27632 and EA+blocking agent Y27632 treatment groups had significantly reduced mRNA and protein expression of Nogo-A, NgR, LINGO-1, RhoA and ROCK II in spinal cord tissues, increased mRNA and protein expression of MLCP, decreased p-MYPT1 protein expression and p-MYPT1/MYPT1 ratio, and caspase3 expression, and improved lower limb movement function after treatment for 14 days (P<0.01 or <0.05). The combination of EA and the blocking agent Y27632 was superior to EA or blocking agent Y27632 treatment alone (P < 0.01 or <0.05).
EA may have an obvious inhibitory effect on the Nogo/NgR and Rho/ROCK signaling pathway after SCI, thereby reducing the inhibition of axonal growth, which may be a key mechanism of EA treatment for SCI.
观察电针(EA)治疗脊髓损伤(SCI)大鼠中Nogo/NgR和Rho/ROCK信号通路相关基因及蛋白表达的变化,进一步探讨EA治疗SCI的可能机制。
采用Allen法制备SCI大鼠模型。64只模型大鼠根据接受的治疗进一步分为4个亚组,即SCI模型组(SCI)、EA治疗组(EA)、阻断剂Y27632治疗组(Y27632)和EA + 阻断剂Y27632治疗组(EA + Y)。对大鼠进行EA和/或阻断剂Y27632治疗。14天后,提取损伤的脊髓组织进行分析。分别通过实时荧光定量PCR和蛋白质印迹法测定mRNA和蛋白表达水平。通过原位杂交评估脊髓中的细胞凋亡变化。采用Basso - Beattie - Bresnahan评估方法评估大鼠后肢运动功能。
与SCI模型组相比,除RhoA蛋白表达外,EA、阻断剂Y27632和EA + 阻断剂Y27632治疗组脊髓组织中Nogo - A、NgR、LINGO - 1、RhoA和ROCK II的mRNA和蛋白表达显著降低,MLCP的mRNA和蛋白表达增加,p - MYPT1蛋白表达及p - MYPT1/MYPT1比值降低,caspase3表达降低,且治疗14天后下肢运动功能改善(P < 0.01或<0.05)。EA与阻断剂Y27632联合应用优于单独使用EA或阻断剂Y27632治疗(P < 0.01或<0.05)。
EA可能对SCI后的Nogo/NgR和Rho/ROCK信号通路具有明显的抑制作用,从而减少对轴突生长的抑制,这可能是EA治疗SCI的关键机制。