Change of m-calpain in transected and sutured sciatic nerve of rat

大鼠坐骨神经横切缝合后m-钙蛋白酶的变化

• 批准号: 12671411
• 负责人: NAKAGAWA Yasuaki
• 金额: $ 2.05万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671411/
• 关键词: Sciatic nerve; transection; suture; Wallerian degeneration; m-calpain; 末梢神経; 再生; mカルパイン

Wallerian degeneration induces neurofilament degeneration in transected peripheral nerve. It has been reported that m-calpain exists in rat sciatic nerve and may play a significant role during Wallerian regeneration. Eight-week-old male Wister rats were obtained. The right sciatic nerve was cut sharply at the postero-inferior portion of right hip joint and the transected ends were retracted to prevent regeneration. The left sciatic nerve which was transected was sutured with 10-0 nylon monofilament with microscope. After sciatic nerves were treated (transected only or sutured), on next day (first day), third day, next week (first week), second week, fourth week, sixth weed, eighth week, twelfth week and sixteenth week, the nerve ends (transected only or sutured) were obtained at proximal 8 mm and distal 8 mm. Each sample was divided into two aliquots, the first aliquot was assayed for protein, the second aliquot was subjected to Western blot analysis. Each sample was normalized by the concentration of protein. In order to quantity the amount of m-calpain at proximal and distal ends of sutured and transected sciatic nerves increased in time course experiments. But in the sutured nerves, the amount of m-calpain at proximal ends decreased after eighth week, and at distal ends tended to diminish after twelfth week. In contrast, in the transected nerves, the tendency could not be seen. In view of many reports of calpain inhibitors as neuroprotectors, during Wallerian degeneration, in early stage, calpain play a positive role, but in late stage, calpain may be an inhibitor of normal regeneration of peripheral nerves.

沃勒氏变性导致周围神经横断后神经丝变性。已有报道称，m-calain存在于大鼠坐骨神经中，可能在沃勒再生过程中发挥重要作用。取8周龄雄性Wistar大鼠。在右侧髋关节后下部切断右侧坐骨神经，并将切断的末端缩回以防止再生。切断左侧坐骨神经，显微镜下用10-0尼龙单丝缝合。分别于第2天(第1天)、第3天、下一周(第1周)、第2周、第4周、第6周、第8周、第12周和第16周取坐骨神经近端8 mm处和远端8 mm处。每个样品被分成两个等份，第一个等份进行蛋白质检测，第二个等份进行蛋白质印迹分析。每个样本通过蛋白质浓度进行归一化。为了定量，在时程实验中增加了缝合和切断坐骨神经近端和远端的m-calain的含量。但在缝合的神经中，近端的m-calain在8周后减少，远端的m-calain在12周后趋于减少。相比之下，在横断的神经中，看不到这种趋势。鉴于目前已有大量的钙蛋白酶抑制剂作为神经保护剂的报道，在沃勒氏变性的早期，钙蛋白酶发挥了积极的作用，但在晚期，它可能是周围神经正常再生的抑制因子。