外源性心脏营养素-1对脊髓损伤后保护性研究 - 第二军医大学学报.docVIP

外源性心脏营养素-1对脊髓损伤后保护性研究 李菁，朱磊林浩东陈爱民 上海[基金项目]上海市局计划项目. Supported by Talent Strategy Program of Shanghais Education Bureau (XYQ020111021). 李菁，男，硕士生，第二军医大学附属长征医院，住院医师。E mail：li0j0i0n0g@126.com 陈爱民，男，博士生corresponding author ）mail：aiminchen@ 【摘要】：目的:探讨外源性心脏营养素-1（Cardiotrophin-1，CT-1）对脊髓损伤的营养保护性作用，分析CT-1及其受体在损伤脊髓中mRNA的表达规律。方法: 取健康雄性SD大鼠90只，体重200－250g。随机分为三组，即空白对照组、神经营养因子（NGF）组及心脏营养素-1(CT-1)组，每组30只。用Allen’s打击法建模，实验组通过导管连续给予人重组心脏营养素（rhCT-1，100μg/kg/d）硬膜内注射，另外两组给予等剂量的NGF及生理盐水，于给药后的第3d、1W、2W对损伤脊髓进行取材。经HE、Nissl染色比较大鼠损伤脊髓神经元及尼氏体数量。另外,再取大鼠60只，经同样方法处理，分实验组与对照组比较1W、4W、12W后去神经营养作用的小腿三头肌湿重及肌肉总蛋白含量，探讨CT-1对脊髓损伤后下肢肌肉功能影响。最后，再取大鼠90只，经同样方法处理，采用荧光定量PCR法检测损伤脊髓中CT-1及其受体亚基gp130、LIFR的mRNA表达。结果:全部切片放大200倍的显微镜视野中进行观察，CT-1组神经元及尼氏体数量与NGF组均无明显统计学差异，且均高于空白对照组。另外，4W、12W时实验组中去神经营养作用的肌肉湿重及蛋白含量均优于对照组。CT-1mRNA的表达于损伤后出现一过性增高，而后呈进行性下降趋势，其受体表达出现反馈性增高。 结论:CT-1（心脏营养素-1）作为一种心肌细胞内广泛表达的细胞因子对脊髓损伤后神经元存活方面具有一定的保护性作用。 脊髓损伤心脏营养素表达规律 The protective effect of exogenous cardiotrophin-1(CT-1) on spinal cord injury LI Jing , ZHU Lei , LIN Haodong , CHEN Aimin Department of Trauma and Reconstruction Surgery , Changzheng Hospital , Second Military Medical University , Shanghai 200003 China 【Abstract】Objective To investigate the protective effect of exogenous cardiotrophin-1(CT-1) on spinal cord injury and analyze the mRNA expression of CT-1 and its receptor. Methods: 90 male rats weight between 200 to 250g were randomly divided into 3 groups. The rats were crashed by Allen’s Weight-dropping Model. The different groups were treated by intraspinal injection of NS(normal saline), NGF, Recombinant human CT-1 (rhCT-1, 100 μg/kg/d),the survival rate and the number of nissl bodies of neurons were compared in HE and Nissl staining after damage 3d, 1w, 2w. Another 60 rats which were denervated for 1w, 4w, 12w by the same model in tow groups were taken to compare wet weight and total protein of surae triceps. Fluorescent quantitative PCR is available to conclude the mRNA expression of CT-1 and its receptor with 90 male rats