The Mechanism Of Angiotensin Ⅱ Induces Neural Stem Cells Differentiated Into Dopaminergic Neurons In Vitro

Objective:To explore the function of Angiotensin type 1 and type 2 (AT1 and AT2)recepters in the possible effects of Angiotensin II induced the differentiation of dopaminegic phenotype neurons from neural stem cells in vitro.Methods:NSCs were isolated from newborn(in 24h) Sprague Dawley rats and cultured in the serum-free medium in vitro. Use the cell grouth culve to describe the capacity of self-newing and proliferation of NSCs. The reproductive activity of NSCs was identify by Nestin immunocytochemical staining. Then, we divided 6 groups, Group A:control, Group B:Aâ…¡, Group C:AT1 antagonist ZD7155, Group D:ZD7155+Aâ…¡, Group E:AT2 antagonist PD123319, Group F:PD123319+Aâ…¡. Differentiated cells were detemined by NSE (neuron special enolase),GFAP (glial fibrillary acidic protein),CNP (cyclic nucleotide phosphohydrolase) immunocytochemical staining to investigate the multi-direction differentiation potency of NSCs. Differentiated cells were detemined by TH (tyrosine hydroxylase) immunocytochemical staining to investigate the process length and Real-Time PCR to detect the expression of TH gene level. Result:NSCs isolated from newborn Sprague Dawley rats grew in the form of neurosheres.We can find Nestin positive in neurosheres. Differentiated cells were detected NSE,GFAP,CNP positive, demonstrated the multi-direction differentiation potency of NSCs. The percentage of NSE+ cells in group B were 10.23%.The ratio of GFAP+ cells in group B (Aâ…¡),D (ZD7155+Aâ…¡,F (PD123319+Aâ…¡) is 49.65%,48.70%,50.97% respectively, which had significantly difference compare with control(44.7%).Real-Time PCR revealed that the TH gene expression of group B (Aâ…¡)and D (ZD7155+AII)were significantly higher then the control group(p<0.05).There were no significant difference in groups of process length of TH positive cells.Conclusion:Exogenous All could promote NSCs differentiated into neurons,astrocytes. All promoted the differentiation of NSCs to DA neurons via AT2 recepter.AT1/AT2 recepter did not involve in the increasing of the length process of TH positive cells.