目的 探讨法舒地尔在改善周围神经损伤后的轴突与髓鞘再生及功能恢复的效果。 方法 SD大鼠30只，重（200±30）g，切断右侧坐骨神经，缝合后等分入2组，即对照组与法舒地尔组，分别给予生理盐水与10mg/kg的盐酸法舒地尔腹腔注射。术后2周，利用NF-200一抗对损伤远端的轴突密度进行评估。术后4周，利用逆行示踪剂荧光金评估发出轴突进入损伤远端的位于L4~6 DRG和腰骶膨大处的神经元数量。术后12周，对腓肠肌的湿重比、肌纤维的横截面积进行测量；利用NF-200与MPZ一抗及透射电镜对轴突的直径与髓鞘的厚度进行测量。术后4、6、8、10、12周采集足印，对两组大鼠坐骨神经功能指数（SFI）进行评定。此外，体外培养大鼠胚胎脊髓背根神经节（DRG）和脊髓运动神经元（SMN），评估法舒地尔对其轴突生长的影响。 结果 术后14 d，法舒地尔组损伤远端神经密度显著高于对照组（P=0.034）。术后4周法舒地尔组被荧光金逆行标志的L4~6 DRG及脊髓前角运动神经元数目均显著高于对照组（P<0.05）。术后12周，法舒地尔组有髓轴突的数量、有髓轴突的直径以及髓鞘的厚度都高于对照组（P<0.05），G-ratio值则低于对照组（P<0.05）。SFI数据结果显示，术后6、8、10及12周，法舒地尔组SFI值显著优于对照组（P<0.05）。术后12周，对照组的患侧腓肠肌的湿重比及肌纤维的横截面积显著小于法舒地尔组（P<0.01）。培养5 d后，法舒地尔组DRG及SMN的轴突显著长于对照组（P<0.01）。 结论 法舒地尔能够促进大鼠坐骨神经横断伤后的轴突与髓鞘再生及其运动功能恢复。
Abstract
Objective To investigate the effect of Fasudil in improving axon and myelin regeneration and functional recovery after peripheral nerve injury. Methods Thirty SD rats weighing (200±30) g were randomly divided into two groups: control group and Fasudil group. Normal saline and 10 mg/kg hydrochloride Fasudil were intraperitoneal given, respectively. The right sciatic nerve was transected and sutured. Two weeks after operation, the density of NF-200 positive axons at the distal end of the injury was evaluated. Four weeks after operation, the number of neurons in the L4-6 dorsal root ganglia (DRG) and sacrolumbar enlargement that sent axons to the nerve distal to the transection site was evaluated by retrograde tracing using Fluoro-gold. Twelve weeks after operation, the wet weight ratio of gastrocnemius muscle and the cross-sectional area of muscle fibers were measured; and immunohistochemistry using primary NF-200 and MPZ antibodies and transmission electron microscopy were used to measure the diameter and thickness of axons and myelin sheaths. The footprints of rats at 4, 6, 8, 10, and 12 weeks after operation were collected to evaluate the sciatic function index (SFI) of rats in the two groups. In addition, rat embryonic dorsal root ganglion (DRG) and spinal motor neurons (SMN) were cultured to evaluate the effect of Fasudil on axon growth. Results At 14 days after operation, the density of NF-200 positive axons at the distal end of the Fasudil group was significantly higher than that of control group (P=0.034). Four weeks after operation, the number of L4-6 DRG and spinal ventral horn motor neurons retrogradely labeled by Fluoro-gold in Fasudil group was significantly higher than that in control group (P<0.05). Twelve weeks after operation, the number of myelinated axons, diameter of myelinated axons, and thickness of myelin sheaths in Fasudil group was higher than those in control group (P<0.05), while G-ratio value was lower than that in control group (P<0.05). The SFI data showed that at 6, 8, 10, and 12 weeks after operation, the SFI values in Fasudil group were significantly better than those in control group (P<0.05). Twelve weeks after operation, the wet weight ratio of gastrocnemius muscle and cross-sectional area of myofibrils in control group were significantly smaller than those in Fasudil group (P<0.01). After culturing for 5 days, the axons of DRG and SMN in Fasudil group were significantly longer than those in control group (P<0.01). Conclusions Fasudil can promote axon and myelin regeneration after sciatic nerve injury in rats and improve their motor function recovery.
关键词
ROCK抑制剂； /
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法舒地尔； /
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坐骨神经横断伤； /
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轴突与髓鞘再生
Key words
ROCK inhibitors; /
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Fasudil; /
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Sciatic nerve injury; /
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Axon and myelin regeneration
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参考文献
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基金
福建省自然科学资金（2021J01241, 2021J01666）
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