摘要
目的:探讨白藜芦醇联合脂肪源性干细胞(ADSCs)分化的施万细胞样细胞(SCLCs)对大鼠坐骨神经损伤的修复作用.方法:ADSCs原代培养并向SCLCs诱导分化,扫描电镜观察细胞形态;Western Blot方法检测S100钙结合蛋白β(S100β)、p75神经营养素受体(p75NTR)、胶质纤维酸性蛋白(GFAP)表达.大鼠随机分为对照组(Control)、施万细胞样细胞组(SCLCs)、白藜芦醇组(Res)、白藜芦醇+施万细胞样细胞组(Res+SCLCs),坐骨神经损伤模型造模成功后8周通过足迹实验检测各组坐骨神经功能指数(SFI);von Frey丝刺激针测定机械缩足阈值(MWT);称重法测定胫前肌湿重比率(WR);Western Blot、RT-qPCR方法检测损伤处神经营养素-3(NT-3)、神经生长因子(NGF)、类胰岛素生长因子-1(IGF-1)和脑源性神经营养因子(BDNF)表达情况.结果:ADSCs诱导8 d后细胞两极细长,细胞外成分增多;S100β、p75NTR、GFAP蛋白高表达.给予SCLCs、Res及 Res+SCLCs 治疗后,治疗组 SFI、WR 明显优于 Control 组(P＜0.05);Res+SCLCs 组、SCLCs 组大鼠 MWT 降低(P＜0.05).Western Blot结果显示:Res+SCLCs组大鼠NT-3、IGF-1、NGF、BDNF蛋白表达高于其余各组(P＜0.05);SCLCs 组大鼠 NT-3、NGF、BDNF 蛋白表达高于 Control 组(P＜0.05);Res 组大鼠 NT-3、NGF 蛋白表达高于 Control 组(P＜0.05).RT-qPCR 结果显示:Res+SCLCs 组大鼠 NT-3、IGF-1、NGF、BDNF mRNA 高表达;SCLCs 组大鼠 NT-3、IGF-1、NGF、BDNF mRNA 表达高于 Control 组(P＜0.05);Res 组大鼠 IGF-1、NGF mRNA 表达高于Control组(P＜0.05).结论:Res联合ADSCs分化的SCLCs对大鼠坐骨神经损伤具有良好的修复作用.
Abstract
Objective:To explore the repair effect of resveratrol combined with Schwann cell-like cells(SCLCs)dif-ferentiated from adipose-derived stem cells(ADSCs)on sciatic nerve injury in rats.Methods:ADSCs were primarily cultured and induced to differentiate into SCLCs.Cell morphology was observed by scanning electron microscopy.West-ern Blot method was used to detect the expressions of S100 calcium-binding protein β(S100β),p75 neurotrophin receptor(p75NTR),and glial fibrillary acidic protein(GFAP).Rats were randomly divided into Control group(Con-trol),Schwann cell-like cell group(SCLCs),resveratrol group(Res),and resveratrol+Schwann cell-like cell group(Res+SCLCs).Eight weeks after the successful establishment of the sciatic nerve injury model,the sciatic nerve func-tion index(SFI)of each group was detected by footprint experiment;the mechanical withdrawal threshold(MWT)was measured by von Frey filament stimulation needle;The wet weight ratio(WR)of the tibialis anterior muscle was deter-mined by weighing method;Western Blot and RT-qPCR methods were used to detect the expressions of neurotrophin-3(NT-3),nerve growth factor(NGF),insulin-like growth factor-1(IGF-1),and brain-derived neurotrophic factor(BDNF)at the injury site.Results:After 8 days of induction of ADSCs,the cells had elongated poles and increased extracellular components;S100β,p75NTR,and GFAP proteins were highly expressed.After treatment with SCLCs,Res,and Res+SCLCs,the SFI and WR of the treatment groups were significantly better than those of the Control group(P＜0.05);the MWT of rats in the Res+SCLCs group and SCLCs group was reduced(P＜0.05).Western Blot re-sults showed that the expressions of NT-3,IGF-1,NGF,and BDNF proteins in rats in the Res+SCLCs group were higher than those in other groups(P＜0.05);The expressions of NT-3,NGF,and BDNF proteins in rats in the SCLCs group were higher than those in the Control group(P＜0.05);the expressions of NT-3 and NGF proteins in rats in the Res group were higher than those in the Control group(P＜0.05).RT-qPCR results showed that the expressions of NT-3,IGF-1,NGF,and BDNF mRNA in rats in the Res+SCLCs group were highly expressed;the expressions of NT-3,IGF-1,NGF,and BDNF mRNA in rats in the SCLCs group were higher than those in the Control group(P＜0.05);The expressions of IGF-1 and NGF mRNA in rats in the Res group were higher than those in the Control group(P＜0.05).Conclusion:Res combined with SCLCs differentiated from ADSCs has a good repair effect on sciatic nerve inju-ry in rats.
维普
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