丹曲林对急性脊髓损伤大鼠脊髓白质的保护作用  

Protective effects of dantrolene on white matter after acute spinal cord Injury in rats

作　　者：瓮调调 廖博[2] 毕一鸣[3] 史明[1] 曾联婷 赵永[1] 付炜[1] 韩军良[1] 

机构地区：[1]第四军医大学西京医院神经内科,西安710032 [2]第四军医大学唐都医院骨科,西安710038 [3]第四军医大学学员一旅,西安710032 [4]汉中市中心医院,汉中723000

出　　处：《神经解剖学杂志》2016年第6期727-731,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81372045)

摘　　要：目的：探讨丹曲林（dantrolene，DA）对大鼠急性脊髓损伤后脊髓白质的保护作用。方法：采用钳夹法制作大鼠急性脊髓损伤模型（SCI）。将48只成年雄性Sprague—Dawley大鼠体重280～310g，随机分成3组，每组16只，假手术组（Sham组）：只行椎板切除术；SCI组：术前1d腹腔注射生理盐水；丹曲林处理组（DA组）：术前1d腹腔注射10ms／kg丹曲林。SCI损伤后3、7、14、21、28d利用BBB评分量表评估大鼠后肢运动功能恢复情况；术后24h，利用抗NF200单克隆抗体的免疫荧光组织化学法标记轴突，观察各组轴突的形态和数量变化；WesternBlot方法检测各组脊髓损伤部位的髓鞘碱性蛋白（Myelin basic protein，MBP）和环核苷酸磷酸二酯酶（2’，3’-cyclic nucleotide phosphodiesterase，CNPase）的蛋白表达水平。结果：不同时间点SCI组大鼠的BBB评分显著低于Sham组，提示SCI模型制备成功。注射丹曲林后，不同时间点SCI动物的BBB评分明显升高（P〈0．05），提示丹曲林可促进大鼠后肢运动功能的恢复。NF200免疫荧光组织化学法显示：Sham组轴突长，排列有序、分布均匀；SCI后24h，NF200’纤维断裂、数量减少、排列紊乱、部分甚至卷曲；给予丹曲林后，NF200＋纤维数量显著增多且较长，排列较规则。Western Blot结果显示：SCI组CNPase，MBP蛋白的表达量较Sham组显著降低（P〈0．01），给予丹曲林后可明显升高两者的表达（P〈0．01）。结论：丹曲林对急性脊髓损伤大鼠白质具有一定的保护作用。Objective： To investigate the protective effect of Dantrolene （DA） on spinal white matter after acute spinal cord injury. Methods： Spinal cord injury （SCI） model was established with clamping methord. Forty-eight adult male Sprague-Dawley rats, weighting 280 - 310 g, were randomly divided into 3 groups （n = 16 each） ： the sham group （the rats only subjected to laminectomy） , the SCI group （the rats received intraperitoneal injection of physiological saline the day before surgery） and the DA group （ the rats received intraperitoneal injection of Dantrolene 10 mg/kg the day before surgery）. BBB rating scale was used to evaluate rat hind limbs movement function recovery in 3, 7, 14, 21 d and 28 d after surgery. NF200, an immunohistochemistry monoclonal antibody of axon, was used to show the morphological and quantitative changes of axon nerve fibers in each group at 24 h after injury. Western Blot was used for analysis of the expression of myelin basic protein （MBP） and 2＇, 3＇-cyclic nucleotide phosphodiesterase （CNPase） of the injured spinal cord in each group. Results： BBB scores in the SCI group were significantly lower than that in the sham group at different time points, indicatinng that SCI model was established successfully. BBB scores were significantly improved after injection of DA at different time point（ P 〈 0.05 ）, indicating that Dantrolene may promote rat hind limbs functional recovery of SCI rats. NF200 immunostaining results showed that the axonal fibers were long, and arranged in an orderly and uniform appearance in the sham group; At 24 h after SCI, NF200＋ fibers were broken, decreased, and arranged randomly some even curly. After injection of DA, the number of NF200＋ fibers were significantly increased, with longer and more regular arrangement. Western Blot showed that the expression of MBP and CNPase was reduced obviously in the SCI group compared with the sham group （P 〈 0.01 ）. Treatment with DA could increase the expression of MBP an

关 键 词：脊髓损伤 丹曲林 BBB评分 白质 大鼠 

分 类 号：R651.2[医药卫生—外科学]