Sciatic nerve preserved by vitrification solution contains Shenfu Injection repaired allogenic nerve defects in a rat model / 中草药

ABSTRACT

Objective: To investigate the feasibility of improving sciatic nerve vitrification preservation using Shenfu Injection in rats. Methods: At -80℃, SPF male SD rat sciatic nerve segments were preserved in the vitrification solution containing Shenfu Injection (0%, 5%, 10%, 30%, groups A, B, C and D) for 4 weeks. The sciatic nerves were observed by transmission electron microscope (TEM), the biological activity of the nerve was detected by laser scanning confocal microscope (LSCM) after Calcein-AM and PI double staining, and the expressions of Bcl-2 and Bax of the sciatic nerves were tested by Western blotting. 10 mm sciatic nerve defect of the Wistar rats (groups A', B', C' and D') was repaired using the corresponding SD rats sciatic nerves preserved for 4 weeks, the gross appearance and sciatic nerve function index (SFI) were detected at different periods after the surgery, the histological and electrophysiological changes of regenerating nerve were detected at 16 weeks post transplantation. Results: The results of TEM showed that demyelination of the sciatic nerve in group A was severe, and in groups B, C and D were mild. The results of LSCM showed that there were strong green fluorescent and weak red fluorescent in groups B, C and D, and weak green fluorescent and strong red fluorescent in group A. The expression of Bcl-2 and Bax of the sciatic nerves, there were significant differences between groups B, C, D and group A, and between groups C, D and group B (P 0.05). The SFI at different periods after the transplantation, the results of electrophysiological detection, the number of myelinated nerve fibers, and the thickness of myelin at 16 weeks after the surgery, there were significant differences between groups C', D' and groups A', B' (P 0.05). At 16 weeks after the surgery, the results by TEM observation showed that there were fewer fibers, smaller diameter, sparse distribution, thinner myelin sheath in group A' and group B', but the number of regenerated fibers was more, the distribution was extensive, and the myelin sheath was thicker in group C' and group D'. Conclusion: Shenfu Injection has protective effect on sciatic nerve preserved by vitrification, and can improve nerve regeneration after allograft in rats.