川芎嗪对脊髓损伤后小鼠神经保护作用的实验研究  

Neuroprotective effect of tetramethylpyrazine on mice after spinal cord injury

作　　者：李姝君 漆国栋 漆伟[2] 杨祝歆 余志娟 江琼[1] LI Shu-jun;QI Guo-dong;QI Wei;YANG Zhu-xin;YU Zhi-juan;JIANG Qiong(Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases,Chongqing Medical University,Chongqing 400016,China;Department of Orthopedic,Chongqing Orthopedic Hospital of Traditional Chinese Medicine,Chongqing 400010,China;Chongqing Jiulongpo Erlang Community Health Service Center,Chongqing 400050,China)

机构地区：[1]重庆医科大学,中医药防治代谢性疾病重庆市重点实验室,重庆400016 [2]重庆市中医骨科医院骨科,重庆400010 [3]重庆市九龙坡区二郎社区卫生服务中心,重庆400050

出　　处：《中国中药杂志》2023年第14期3848-3854,共7页China Journal of Chinese Materia Medica

基　　金：重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0516);重庆医科大学中医药防治代谢性疾病重庆市重点实验室开放课题(2021-1-7)。

摘　　要：探讨川芎嗪对脊髓损伤后小鼠的神经保护作用及机制。75只雌性C57BL/6小鼠随机分为5组,假手术组、模型组、川芎嗪低剂量组(25 mg·kg-1)、川芎嗪中剂量组(50 mg·kg-1)、川芎嗪高剂量组(100 mg·kg-1),每组15只,采用改良的Rivlin法建立小鼠急性脊髓损伤模型。川芎嗪干预14 d后,采用脊髓损伤评分标准(basso mouse scale,BMS)评分法及斜板实验对小鼠后肢运动功能进行评价。酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测脊髓匀浆中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和白细胞介素-1β(interleukin-1β,IL-1β)的水平。苏木素-伊红(hematoxylin-eosin,HE)染色观察脊髓组织学情况,尼氏染色观察神经元数目变化。通过蛋白免疫印迹法与免疫荧光法检测脊髓组织胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和补体C3蛋白的表达。川芎嗪治疗组对于小鼠脊髓损伤后的后肢运动功能有明显的改善,其中川芎嗪高剂量组BMS评分及斜板实验评分较模型组显著升高(P<0.01)。川芎嗪高剂量组脊髓匀浆中TNF-α、IL-6和IL-1β水平显著降低(P<0.01)。经川芎嗪治疗后脊髓形态有恢复,尼氏体数目明显增加且形态规则,神经元丢失减少。与模型组相比,川芎嗪高剂量组GFAP、C3蛋白表达明显减少(P<0.05,P<0.01)。综上,川芎嗪能促进脊髓损伤后小鼠运动功能的改善,发挥神经保护作用,其机制可能与抑制炎症反应、改善胶质瘢痕的过度增生有关。This study aims to investigate the neuroprotective effect of tetramethylpyrazine on mice after spinal cord injury and its mechanism.Seventy-five female C57BL/6 mice were randomly divided into 5 groups,namely,a sham operation group,a model group,a tetramethylpyrazine low-dose group(25 mg·kg-1),a tetramethylpyrazine medium-dose group(50 mg·kg-1),and a tetramethylpyrazine high-dose group(100 mg·kg-1),with 15 mice in each group.Modified Rivlin method was used to establish the mouse model of acute spinal cord injury.After 14 d of tetramethylpyrazine intervention,the motor function of hind limbs of mice was evaluated by basso mouse scale(BMS)and inclined plate test.The levels of inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in the spinal cord homogenate were determined by enzyme-linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)staining was used to observe the histology of the spinal cord,and Nissl's staining was used to observe the changes in the number of neurons.Western blot and immunofluorescence method were used to detect the expression of glial fibrillary acidic protein(GFAP)and C3 protein.Tetramethylpyrazine significantly improved the motor function of the hind limbs of mice after spinal cord injury,and the BMS score and inclined plate test score of the tetramethylpyrazine high-dose group were significantly higher than those of the model group(P<0.01).The levels of TNF-α,IL-6,and IL-1βin spinal cord homogenate of the tetramethylpyrazine high-dose group were significantly decreased(P<0.01).After tetramethylpyrazine treatment,the spinal cord morphology recovered,the number of Nissl bodies increased obviously with regular shape,and the loss of neurons decreased.As compared with the model group,the expression of GFAP and C3 protein was significantly decreased(P<0.05,P<0.01)in tetramethylpyrazine high-dose group.In conclusion,tetramethylpyrazine can promote the improvement of motor function and play a neuroprotective role in mice after spinal cord inj

关 键 词：脊髓损伤 川芎嗪 神经保护 炎症反应 胶质瘢痕 

分 类 号：R285.5[医药卫生—中药学]