ABSTRACT
Objective To study the effect and mechanism of Akt during Islet -1 inducing the mesenchymal stem cells of C3H10T 1/2 cells to differentiate specifically into cardiomyocytes.Methods Developing a model of Islet-1 over-expression cells, Lentivirus infection efficiency of cells was detected by flow cytometry detection tech-nology.Cell proliferation was tested by CCK-8.The protein expression of Islet-1, cTnT and p-Akt/T-Akt was measured by Western blot.The mRNA expression of GATA4,Nkx2.5 and Mef2c were tested by real-time PCR. Results With the increasing of MK-2206 concentration, the inhibition rate of cell proliferation increased ( P<0.05) ,and the best inhibition concentration of Akt was 8nmol /L; With prolongation of Islet-1 inducing time,the protein expression of p-Akt/TA-kt reduced ( P<0.05 );The gene expression of myocardial specific transcription factor GATA4, Nkx2.5 andMe f2c increased( P<0.05); treated with MK-2206, the geneexpres-sion of GATA4, Nkx2.5 and Mef2cincreased significantly at the first week and then reduced ( P<0.05) . Conclu isons Akt plays different effects in different differentiation stages during the Islet-1 inducing the cells into cardiacs-pecific differentiation process .
ABSTRACT
Objective To explore whether Islet-1 gene of the rat could induce NSCs to differentiate into cholinergic neurons. Methods NSCs were transduced with Islet-1 recombinant retroviral expression vector.The protein expression of Islet-1 gene in NSCs was detected by immunofluorescence histochemical method.The ability of NSCs to differentiate into ChAT positive cells was observed in vivo and in vitro. Results The numbers of ChAT positive cells were significantly increased in the group of NSCs modified with Islet-1 compared with the control group in vitro.NSCs modified with Islet-1 could differentiate into ChAT positive cells when they were grafted into the corpus striatum of the adult rat brain.Conclusion Islet-1 gene could induce NSCs to differentiate into cholinergic neurons.