The Significance Of Notch And Neurogulin-1/ErbB Signaling Pathway In Differentiation Of Schwann-like Cells In Vitro

Objective: To investigate the role of Notch signaling pathway and Neurogulin-1/Erb B signaling pathway in the differentiation of rat bone marrow mesenchymal stem cells(MSCs)into Schwann cells(SCs).Methods: BMSCs were isolated from high glucose medium and cultured in fetal bovine serum(FBS)and ?-MEM medium.The cells(Notch signaling)were stained with ?-mercaptoethanol(?-ME),RA,PDGF-AA,b FGF,Forskolin,Heregulin were divided into three groups: control group,induction group and blocker group.The Notch pathway blocker DAPT was added at induction.The expression of Jagg1 ligand,Hes1 target gene,Notch1,Dll1 receptor protein and signal protein(S100,p75,GFAP)were determined by quantitative RT-q PCR.Cell proliferation was measured by CCK-8.The apoptosis of MSCs was detected by flow cytometry.Neurogulin-1/Erb B pathway experiments were divided into induced Schwann-like cells group,Mubritinib added group,and Heregulin removed during differentiation group.Immunofluorescence technique and flow cytometry were used to detect the cell differentiation level and ratio.Cell proliferation was detected by CCK-8.Cell apoptosis was detected by flow cytometry.Results: Compared with the control group,the expression of SCs signaling protein(S100,p75,GFAP)was significantly increased in the full inducer group and the blocker group;Jagg1 ligand in the blocker group was significantly higher than that of the full inducer group.The receptor protein of Hes1 target gene,Notch1 and Dll1 was significantly decreased.Compared with the whole inducer group,the proliferative effect and apoptosis rate of the blocker group were significantly lower than that of the other two groups(P <0.05)..In the study of the Neurogulin-1/Erb B pathway,it was shown that Erb B2 and Erb B3 did not occur during the initial induction period,and that the expression amount gradually increased with the prolongation of the induction time;real-time fluorescence quantitative PCR results showed that blockage of Neurogulin-1 with a blocker was shown.The degree of differentiation of stem cells after /Erb B signaling pathway was significantly lower than that of the induced group(P<0.05).At the end of differentiation,compared with the inducer group,the cell proliferation was significantly decreased after blocking Erb B2.The early apoptotic rate increased significantly and the difference was significant(P<0.05).Conclusion: 1.inhibition of Notch signaling pathway can inhibit the differentiation of MSCs to SCs,and its mechanism may be related to Notch signaling pathway promote cell proliferation.2.In the process of differentiation of Schwann-like cells from BMSCs,the Neurogulin-1/Erb B signaling pathway has a promoting effect,and the effect is gradually enhanced.The mechanism may be related to the promotion of cell proliferation and promote the early apoptosis of MSCs.