Effect of Lycium barbarum polysaccharide on sciatic nerve injury in rats
Objective To investigate the effect of lycium barbarum polysaccharide(LBP)intervention on Schwann cells(SCs)combined with allogeneic nerve grafts(ANX)in repairing sciatic nerve defects in rats.Methods The sciatic nerve SCs from rats were obtained and primary culture until the third generation was performed.The0,50,100,150,200,250,500 mg/L LBP and SCs were conducted mixed cultivation and divided into 7 groups.On the 1st,3rd,5th,7th,and 9th day after cultivation in culture dishes,the biological activity of each group of SCs was detected using the CCK-8 method.The expression levels of brain derived neurotrophic factor(BDNF),nerve growth factor(NGF),and basic fibroblast growth factor(bFGF)in each group were detected by rt-PCR,and the optimal concentration of LBP for intervention in SCs was screened.The rat model of 10 mm sciatic nerve defect was established under sterile conditions,the tibial nerve of rabbits was selected to make ANX,and SD rats were divided into three groups,with 10 rats in each group:ANX group,ANX-SCs group,and ANX-LBP-SCs group.After 8 weeks of surgery,the sciatic nerve function index(SFI)and motor nerve conduction velocity(MCV)were measured,and the expression level of Caspase-3 in the regenerated nerve was detected using rt-PCR.Additionally,the nerve fiber and myelin sheath regeneration in the nerve grafts of the three models were compared using transmission electron microscopy.Results The activity of SCs at different time points in each well was measured using CCK-8.On the 2nd to 4th day,there was no statistically significant difference in activity among the 7 groups(P＞0.05),while on the 6th to 8th day,the proliferation activity of 150 to 250 mg/L LBP-SCs group was stronger than that of other concentration groups,and the difference was statistically significant(P＜0.05).The mRNA expression levels of BDNF,NGF,and bFGF in the 200 mg/L LBP-SCs group were detected by q-PCR,and the differences were statistically significant compared to other groups(P＜0.05).After 8 weeks of transplantation,it was found that the ANX-LBP-SCs group had better data in thermal stimulation test,SFI detection,and nerve conduction velocity,and the expression level of Caspase-3 was the lowest.The results of transmission electron microscopy showed that the ANX-LBP-SCs group had better degree of myelin edema and nerve fiber regeneration than the other groups.Conclusion The LBP at a concentration of 200 mg/L can promote the proliferation of SCs and secrete a large amount of neurotrophic factors.The SCs culture system with LBP intervention combined with ANX transplantation can significantly promote peripheral nerve regeneration.
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