维拉帕米对脊髓损伤小鼠的神经保护作用  

Neuroprotective effect of verapamil on spinal cord injury in mice

作　　者：郏振杰 徐传华[1] 丁棚辉 JIA Zhenjie;XU Chuanhua;DING Penghui(Department of Pain Treatment,Taizhou Municipal Hospital,Taizhou 318000,CHINA)

机构地区：[1]台州市立医院疼痛科,浙江318000

出　　处：《江苏医药》2022年第10期977-981,F0002,共6页Jiangsu Medical Journal

基　　金：浙江省医药卫生科技计划项目(2020KY367);台州市社会发展科技计划项目(21ywa39)。

摘　　要：目的探讨维拉帕米对脊髓损伤小鼠的神经保护作用及潜在机制。方法将72只C57BL/6小鼠随机均分为三组,每组24只。VP组和NaCl组采用改良Allen’s方法制备脊髓损伤模型;VP组术后予尾静脉注射维拉帕米2 mg·kg^(-1)·d^(-1),连续给药7 d。Sham组与NaCl组予注射等量生理盐水,连续注射7 d。采用Basso-Mouse量表(BMS)评估小鼠后肢运动功能恢复情况,HE染色及免疫荧光染色观察脊髓损伤后组织结构的改变,Western blot、实时荧光定量PCR和ELISA检测小鼠炎症和凋亡相关蛋白或mRNA的表达。结果脊髓损伤后小鼠运动功能减弱,VP组小鼠术后第7和14天的BMS评分高于NaCl组(P<0.05或P<0.01)。HE染色结果显示,VP组较NaCl组脊髓组织的坏死面积减少,组织结构改善。免疫荧光染色结果显示,VP组微管相关蛋白2表达高于NaCl组(P<0.05)。与NaCl组相比,VP组小鼠血清炎症标记物TNF-α、IL-1β水平和脊髓组织凋亡标记物Caspase-3、Bax蛋白和mRNA表达降低(P<0.05),细胞质NF-κB蛋白表达升高,细胞核NF-κB蛋白表达降低(P<0.01)。结论维拉帕米能抑制炎症反应和神经元凋亡,从而改善脊髓损伤后的组织损伤,促进神经功能恢复;其机制可能与抑制NF-κB途径有关。Objective To investigate the neuroprotective effect and underlying mechanism of verapamil on spinal cord injury(SCI)in mice.Methods Seventy-two C57 BL/6 mice were equally randomized into three groups.The SCI model was established by the modified Allen’s weight drop method in groups of A and B.Group B was injected with verapamil 2 mg·kg^(-1)·d^(-1) via caudal vein after operation daily for 7 days.The groups of A and C were taken as the model control and sham operation,which were injected with the same volume of normal saline.Basso-Mouse Scale(BMS)was used to evaluate the recovery of hindlimb motor function.HE staining and immunofluorescence staining were used to explore the changes in the organizational structure after SCI.The expressions of inflammatory and apoptotic proteins or mRNA were determined by Western blot,real time fluorescent quantitative PCR and ELISA.Results The motor function of mice after SCI was decreased.The BMS score in group B was higher than that in group A on the 7^(th) and 14^(th) day after operation(P<0.05 or P<0.01).HE staining showed that compared with group A,the necrosis area of spinal cord tissues was reduced and the tissue structure was improved in group B.Immunofluorescence staining showed that the expression of microtubule associated protein-2 in group B was higher than that in group A(P<0.05).Moreover,compared with group A,serum levels of inflammatory markers(TNF-αand IL-1β)and the expressions of apoptosis markers(Caspase-3 and Bax)in the spinal cord were decreased(P<0.05),cytoplasmic NF-κB expression was increased,and nuclear NF-κB expression was decreased in group B(P<0.01).Conclusion Verapamil can inhibit inflammatory reaction and neuronal apoptosis,improve tissue injury after SCI and promote the recovery of neural function.The mechanism for this may be related to the inhibition of NF-κB pathway.

关 键 词：维拉帕米 脊髓损伤 炎症 凋亡 核因子κB途径 

分 类 号：T651[一般工业技术]