葡萄糖原合成酶-3β抑制剂促进大鼠脊髓损伤后轴突再生与下肢功能恢复的研究  

Glycogen synthase kinase--3β inhibitor （TDZD-8） promotes axonal regeneration and functional recovery of hind limbs after spinal cord injury in rats

作　　者：雷飞[1] 叶飞[1] 冯大雄[1] 康建平[1] 黄贻泽[1] 张戈[1] 屈一鸣[1] 周际[1] 

机构地区：[1]泸州医学院附属医院脊柱外科,四川省泸州市646000

出　　处：《中华创伤骨科杂志》2013年第1期65-71,共7页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（30872602）

摘　　要：目的探讨葡萄糖原合成酶-3β（GSK-3β）的抑制剂噻二唑-8（TDZD-8）对大鼠脊髓损伤再轴突再生与下肢功能恢复的影响。方法健康成年雌性SD大鼠108只，均制作T。平面完全性截瘫的脊髓损伤动物模型，并L4平面蛛网膜下腔插入注药导管后随机分为3组（n=36）：TDZD-8治疗绀（A组）经注药导瞥注入1mg／（kg·d）的TDZD-8，PBS治疗组（B组）经注药导管注入60μL／d的磷酸盐缓冲液，C组为奈门对照组。各组均在瘫痪后1h开始注射，连续注射3周。注射后2h、4h、8h、24h、7d,FUNEL法观察神经元细胞捌亡的表达；注射后7、14、28d，兔抗生长相关蛋白一43（GAP一43）的表达；注射后1、2、3、6、8、12周，Basso—Beattie—Bresnahan（BBB）评分评估下肢运动功能恢复情况；注射开8周，采用免疫荧光法观察损伤节段轴突再生情况。结果脊髓损伤后2h观察到凋亡细胞，注射后2h、4h、8h、2411、7d，A组的神经元细胞凋亡数明显低于B、C组，各组细胞凋亡表达在注射后8h达到岛峰，此后逐渐减少，羞异均有统计学意义（P〈0．05）；注射后7、14、28d，A组的GAP．43表达面积明显高于B、C组．各组GAP-43表达在损伤后14d达到高峰，此后逐渐减少，差异均有统计学意义（P〈0．051；从注射行3周开始，A组的BBB评分明显高于B、C组，差异有统计学意义（P〈0．05）。A组有较多冉生神经纤维穿过损伤部位，而B、C组只有少许再生神经纤维穿过损伤部位，注射后8周A组阳性标记的皮质脊髓束纤维面积明显高于B组、C组，差异均有统计学意义（P〈0．05）。结论 FDZD-8能够抑制细胞凋亡，减少继发性脊髓损伤，上调GAP-43的表达，促进轴突再生、延长，促进大鼠后肢功能恢复。Objective To explore the effect of glycogen synthase kinase-3β（GSK-3β） inhibitor （TDZD-8） on axonal regeneration and functional recovery of hind limbs after spinal cord inju （SCI） in rats. Methods Animal models of complete paraplegia at T9 level were made in 108 aduh female Sprague-Dawley rats which were subjected to weight-drop impact before they were randomly divided into even 3 groups, Group A was trealed with 1 mg/（kg ＂ d） TDZD-8, group B with 60L/d PBS and group C was set as blank controls. All agents were respectively injected through subarachnoid catheters implanted via L4 at one hour after SC1 and the injections lasted for 3 weeks. The expressions of GAP-43 were measured respectively by SP immunohistoehemical staining on 7, 14 and 28 days after injection. The expressions of neuronal apoptosis were observed respee｜ively by TUNEL staining at 2, 4, 8, 24 hours and 7 days after injection. The motor function of hind limbs in each group was evaluated by Basso-Beattie-Bresnahan （BBB） scores at 1, 2, 3, 6. 8 and 12 weeks respectively alter injection. Axonal regeneration was evaluated by immunofluorescence staining at 8 weeks respeelively after injection. Results The expressions of neuronal apoptosis were detected at 2 hours after SCI, and rose to the peak at 8 hours before they gradually decreased. The apoptotic cells in group A were significantly lower than in groups B and C （ P 〈 0.05） at all time points. The expressions of GAP-43 were increased gradually after SCI and rose to the peak on the 14 days before they gradually decreased. The GAP-43 expressions in group A were significantly higher than in groups B and C at all time points （ P 〈 0. 05）. The BBB scores in group A were significantly higher than in groups B and C from 3 weeks after injection （ P 〈 0.05）. More regeneration neurofibers extended through the lesion area in group A while few neurofibers did in groups B and C. At 8 weeks after injection, the positive fiber area of corticospinal tracts was significan

关 键 词：脊髓损伤 酶抑制剂 细胞凋亡 功能恢复 

分 类 号：R651.2[医药卫生—外科学]