睫状神经营养因子促进自体去神经带血管移植肌神经的再生

Ciliary neurotrophic factor in promoting muscular neurotization of denervated vascularized skeletal muscle

目的:探讨睫状神经营养因子在移植肌神经再生过程中的作用。方法:实验于2001-03/2002-03在解放军总医院实验动物中心进行,取SD大鼠36只随机分为对照组和睫状神经营养因子组各18只。两组均切除2个后肢腓总神经各1.5cm,将腓骨长肌远端切断并切除肌膜后植入同侧腓肠肌。睫状神经营养因子组从手术当天开始向每单后肢移植肌注射睫状神经营养因子0.05μg/d,对照组注射生理盐水0.5mL,共7d。通过术后不同时间的组织学、神经髓鞘、神经纤维、乙酰胆碱酯酶染色,神经生长因子抗体、脑源性神经生长因子抗体检测及电子显微镜检查,观察神经再生情况。结果:组织学及神经髓鞘及神经纤维染色显示睫状神经营养因子组在术后不同时间的肌纤维及神经纤维病变等多方面较对照组轻。90d时恢复较对照组好。电镜观察见睫状神经营养因子组无论在肌细胞形态,核仁清晰度,染色质丰富方面均较同期对照组好,可见多处神经再生。乙酰胆碱酯酶染色结果显示:睫状神经营养因子组较对照组有更多的运动终板。脑源性神经生长因子抗体染色显示神经生长因子组较对照组强度更大。神经生长因子抗体染色显示睫状神经营养因子组较对照组强度更大。结论:外源性的睫状神经营养因子对移植肌神经再支配有促进作用,对失神经肌肉有一定的保护作用。

AIM:To study the effects of ciliary neurotrophic factor in the process of muscular neurotization of denervated vascularized skeletal muscle. METHODS:The experiment was performed in the Animal Center of General Hospital of Chinese PLA between March 2001 and March 2002.Totally 36 SD rats were divided randomly into ciliary neurotrophic factor and control groups(n=18).Two common peroneal nerves of 1.5 cm were resected in hind limbs of rats.Gastrocnemii were embedded following resection of sarcolemmas of peroneus longus muscles.From the operation day to the 7th day after operation,daily dose of ciliary neurotrophic factor(0.05 μ g/per hind limb),and 0.5mL saline were injected into the operative site in ciliary neurotrophic factor and control groups, respectively.At various times after operation,the regeneration of nerve was examined with histology,neuronal myelin sheath,nerve fiber,Acetylcholinesterase,nerve growth factor antibody,brain derived neurotrophic factor antibody staining,electron microscope and immunohistochemical methods. RESULTS:Histology,neuronal myelin sheath and nerve fiber staining indicated that the lesion in muscle fiber and nerve fiber was milder in ciliary neurotrophic factor group as compared with control group.At day 90,the recovery in ciliary neurotrophic factor group was better than that in control group.The electron microscope showed better muscle cell form,acuity of nucleolus,more plenty chromatin,and more regeneration of nerve in ciliary neurotrophic factor group.Acetylcholinesterase staining indicated more motor end plates,brain derived neurotrophic factor antibody staining indicated more intensity of nerve growth factor,and nerve growth factor antibody staining indicated more intensity of ciliary neurotrophic factor in ciliary neurotrophic factor group as compared with control group. CONCLUSION:Exogenous ciliary neurotrophic factor can promote the muscular neurotization and diminish denervated muscle atrophy.