吡拉西坦通过MAPK通路治疗大鼠脊髓损伤的疗效观察与机制研究  

Therapeutic effect and mechanism of piracetam for the treatment of spinal cord injury in rats through MAPK pathway

作　　者：董博[1] 李越 李迎春 王桐 梁壮 贺西京[1] DONG Bo;LI Yue;LI Ying-chun;WANG Tong;LIANG Zhuang;HE Xi-jing(Department of Orthopaedics,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710048,Shaanxi,China;The First Clinical College of Medicine,Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,Shaanxi,China)

机构地区：[1]西安交通大学第二附属医院骨科,陕西西安710048 [2]陕西中医药大学第一临床医学院,陕西咸阳712000

出　　处：《中国骨伤》2024年第6期591-598,共8页China Journal of Orthopaedics and Traumatology

基　　金：陕西省重点研发计划项目“两链”融合重点专项(编号:2021LLRH-08);陕西省教育厅青年创新团队计划(编号:22JP018);秦都区2022年科技计划项目(编号:QK2022sf04)。

摘　　要：目的:探讨吡拉西坦通过丝裂原激活蛋白激酶(mitogen-activated protein kinase,MAPK)通路治疗脊髓损伤大鼠的作用机制。方法:将54只体重为80~100 g的6周龄SD雌性健康大鼠采用随机数字表法分为假手术组、脊髓损伤组、吡拉西坦组,每组18只。脊髓损伤组、吡拉西坦组使用打击器建立脊髓损伤模型,假手术组不损伤脊髓。吡拉西坦组按照5 ml·kg-1标准予尾静脉注射吡拉西坦,连续干预3 d,每日1次;其他2组注射等剂量、等次数、等时长的生理盐水。分别于术后1、3、7 d处死大鼠并取材,观察并比较大鼠脊髓损伤行为学评分(Basso,Beattie and Bresnahan locomotor rating scale,BBB)的变化,使用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测脊髓炎症因子白细胞介素-6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)、白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平变化,HE染色观察脊髓损伤大鼠的形态学变化,免疫组化观察水通道蛋白-4(aquaporin 4,AQP4)表达水平,蛋白质免疫印迹法(western blotting,WB)观察脊髓损伤后MAPK信号通路在大鼠脊髓中的激活状态。结果:假手术组1、3、7 d的BBB评分均为21分;脊髓损伤组分别为(1±1)、(4±1)、(7±2)分;吡拉西坦组分别为(1±1)、(5±1)、(9±2)分;脊髓损伤组与假手术组比较,差异有统计学意义(P<0.05);吡拉西坦组与脊髓损伤比较,差异有统计学意义(P<0.05)。HE染色结果显示,假手术组形态未见异常。脊髓损伤组在术后1 d时,脊髓组织出现出血、变性;术后3 d,脊髓组织内出现片状坏死区;术后7 d,脊髓组织开始缓慢修复。吡拉西坦组术后1 d,脊髓损伤出血面积较脊髓损伤组减小;术后3 d,坏死区域较脊髓损伤组减少,细胞核消失范围较脊髓损伤组缩小;术后7 d,脊髓开始缓慢恢复。假手术组大鼠造模后1、3、7 d脊髓组织中AQP4染色较淡;脊髓损伤Objective To explore mechanism of piracetam for the treatment of spinal cord injury in rats through mitogen-activated protein kinase(MAPK)pathway.Methods Fifty-four healthy 6-week-old SD female rats with body weight of 80 to 100 g were divided into sham operation group,spinal cord injury group and piracetam group by random number table method,with 18 rats in each group.Spinal cord injury model was established in spinal cord injury group and piracetam group using percussion apparatus,while sham operation group did not damage spinal cord.Piracetam group was injected with piracetam injection through tail vein according to 5 ml·kg^(-1)standard,once a day for 3 days;the other two groups were injected with normal saline at the same dose,the same frequency and the same duration.The rats were sacrificed at 1,3,and 7 days after surgery,and changes of Basso,Beattie and Bresnahan(BBB)locomotor rating scale was observed and compared.Enzyme-linked immunosorbent assay(ELISA)was used to detect spinal cord inflammatory factors,such as interleukin-6(IL-6),interleukin-10(IL-10),interleukin-1β(interleukin-1β),necrosis factor-α(IL-1β)and tumor necrosis factor-α(TNF-α);HE staining was used to observe morphological changes of rats with spinal cord injury,and immunohistochemistry was used to observe expression level of aquaporin 4(AQP4).The activation of MAPK signaling pathway in spinal cord of rats after spinal cord injury was observed by western blotting(WB).Results BBB scores of sham operation group on 1,3 and 7 day were 21 points.In spinal cord injury group,the scores were(1±1),(4±1)and(7±2);piracetam group was(1±1),(5±1),(9±2),respectively;the difference between spinal cord injury group and sham operation group was statistically significant(P<0.05).HE staining showed that no abnormality was found in sham operation group.In spinal cord injury group,bleeding and degeneration of spinal cord tissue appeared at 1 day after operation;flaky necrotic areas were appeared in spinal cord at 3 days after surgery,and spinal cord t

关 键 词：吡拉西坦 脊髓损伤 MAPK信号通路 

分 类 号：R741[医药卫生—神经病学与精神病学]