Effect Of Faslg On Nerve Degeneration And Regeneration After Rat Sciatic Nerve Injury

ObjectiveBy using cell biology technology,molecular biology technology and bioinformatic analysis,we investigated the molecular mechanism of Faslg on Degeneration and Regeneration after Rat Sciatic Nerve Injury.Wallerian is an anterograde degeneration in the distal end after peripheral nerve injury.Previous studies have shown that Faslg plays an important role in the Wallerian.This study aims to explore the effect of Faslg on cell proliferation,cell migration,cell apoptosis and other functions of SCs in the Wallerian after Rat Sciatic Nerve Injury,and explore the molecular mechanism that Faslg regulated on Degeneration and Regeneration after Sciatic Nerve Injury,the study provided more theoretical basis for us on the molecular mechanisms for repairing the nerve injury and functional recovery.Methods1.To establish the rat sciatic nerve transection injury models 2.Immunocytochemistry(ICC)and Immunohistochemistry(IHC)analysis the location of Faslg 3.Faslg si RNA and overexpression plasmid transfection into SCs in vitro 4.Real-time PCR analysis5.Western Blot analysis 6.Ed U proliferation assay,Annexin V apoptosis assay,and Transwell migration assay 7.Functional experiment in vivo,to test the function of Faslg in vivo 8.Using TUNEL to detect the apoptosis in the tissue 9.Using SPSS statistical analysis software for the single factor analysis of variance,all data were represent as standard SD and mean (?)((?)± SD)and P < 0.05 for results with statisticalsignificance.Results1.ICC and IHC analysed the co-location of Faslg and S100 B in SCs.2.After the rat sciatic nerve injury,the expression of Faslg significantly increased in degeneration and regeneration.3.The number of SCs proliferation and migration increased after Faslg si RNA interference,but the number of apoptotic cells decreased.4.The number of SCs proliferation and migration decreased after Faslg Plasmid overexpression,but the number of apoptotic cells increased.5.The expressions of related-factors have changed after Faslg si RNA interference and Plasmid overexpression.6.After Faslg si RNA interference and Plasmid overexpression in SCs,the protein expression of related signal pathway such as c-fos,p-c-jun/c-un,N-Ras have unchanged,but ?-catenin,p-ERK/ERK,p-AKT/AKT,NF-?B,Caspase-3 are markedly increased.7.After Faslg si RNA interference and Plasmid overexpression in vivo,the protein expression of related signal pathway such as p-ERK/ERK,c-fos,p-c-jun/c-jun,NF-?B,N-Ras have not significantly stable changes,but ?-catenin,p-AKT/AKT,Caspase-3 are markedly changed.8.After Faslg si RNA interferencein vivo,the number of cells apoptosis decreased during the repair after sciatic nerve injury;but the number of cells apoptosis did not change after Faslg overexpression.Conclusions1.The co-location of Faslg and S100 B indicates that Faslg expresses on the SCs membrane 2.Faslg inhibits the proliferation and migration of SCs,but promotes the apoptosis of SCs 3.Faslg may regulate the regeneration and repair of sciatic nerve injury by ?-catenin,p-ERK/ERK,p-AKT/AKT,NF-?B and Caspase-3.4.In vivo,Faslg may regulate the regeneration and repair of sciatic nerve injury by ?-catenin,p-AKT/AKT and Caspase-3.5.Faslg plays a role in cell apoptosis on nerve degeneration and regeneration after rat sciatic nerve injury.