人参皂甙Rb1冷保存大鼠坐骨神经修复周围神经缺损的实验研究

Experimental study of ginsenoside Rb1 on cold-preserved sciatic nerve and nerve regeneration in a rat model

[目的]探讨人参皂甙Rb1(Ginsenoside Rb1,G-Rb1)冷保存大鼠坐骨神经对同种异体移植后受体神经再生的影响。[方法]3个月龄SPF级雄性Wistar大鼠和SD大鼠分别作为供体和受体。切取供体大鼠双侧坐骨神经15 mm长,随机在G-Rb1溶液(0、50、200 mg·L-1)中4℃保存4周(A4、B4、C4组)和6周(A6、B6、C6组),Western-blot检测供体神经Bcl-2表达,Calcein-AM染色LSCM观察冷保存6周供体神经生物活性。用冷保存6周的供体神经(A6、B6、C6组)修复对应的受体(A、B、C组)坐骨神经10 mm缺损,术后分期行大体观察、坐骨神经功能指数检测,第20周行电生理检测、再生神经组织学观察。[结果]Western-blot检测Bcl-2表达,冷保存4周时,B4组高于A4、C4组(P<0.05),冷保存6周时,B6、C6组高于A6组,且B6组高于C6组,差异均有统计学意义(P<0.05)。Calcein-AM染色LSCM观察,B6、C6组荧光强度与A6组比较,B6组与C6组比较,差异均有统计学意义(P<0.05)。移植术后各期坐骨神经功能指数,20周电生理检测、再生有髓神经纤维数目、髓鞘厚度,B、C组与A组比较,B组与C组比较,差异有统计学意义(P<0.05);术后20周透射电镜显示,A组再生有髓纤维少、髓鞘薄,结缔组织增生明显,B、C组有髓纤维较多、髓鞘厚,无明显结缔组织增生。[结论]G-Rb1对冷保存大鼠坐骨神经具有保护作用,G-Rb1冷保存的坐骨神经能改善同种异体移植后神经再生效果。

[Objective]This study aimed to investigate the effects of Ginsenoside Rb1 on cold- preserved sciatic nerve and recipients＇ nerve regeneration after transplantation in rats. [Method]SPF male Wistar rats and SD rats（ 3 months old） were selected as donor and recipient respectively. The sciatic nerve segments of 15 mm were harvested from the donors and then randomly preserved in 4℃ Ginsenoside Rb1 solution（ 0,50,200 mg / L） for 4 weeks（ groups A4,B4,C4） and 6 weeks（ groups A6,B6,C6）. The expression of Bcl- 2 protein of the nerves were detected by Western- blot,the biological activity of the nerve preserved for 6 weeks was observed by laser confocal microscope（ LSCM） with calcein- AM staining. The defect of recipients＇ sciatic nerves（ 10 mm） was repaired by the corresponding donor ＇s nerve preserved for 6 weeks,the gross appearance and sciatic nerve function index（ SFI） were detected at different period after the surgery,and the electrophysiological and histological changes of regenerated nerve were observed at 20 weeks after transplantation. [Result]After preserved for 4 weeks,the expression of Bcl- 2 protein in group B4 was higher than that in groups A4 and C4（ P〈 0. 05）,and after preserved for 6 weeks,Bcl- 2 expressions in groups B6andC6 were higher than that in group A6,and in group B6 was higher than in group C6（ P 〈0. 05）. For the fluorescence intensity observed by LSCM,there were significant differences among three groups（ A6,B6,C6）（ P〈 0. 05 =. The There were significant differences among the three groups（ A,B,C） considering the SFI at different time points after transplantation,the electrophysiological results as well as the number of myelinated nerve fibers and thickness of myelin at 20 weeks（ P 0. 05 =. The transmission electron microscopy results at 20 weeks showed that thickness of myelin sheath in group A was thinner than those in group B and C,and connective tissue hyperplasia in group A was more serious than those in group B and C. [Conclusion]G- Rb1 plays a protective role on cold preservation of sciatic nerve,and it can improve nerve regeneration after nerve transplantation.