RESUMO
Objective To study the effect of Nurrl gene on the differentiation of rats' marrow stromal cells(MSCs) into neurous under the co-inducement of total panax notoginserg saponins(tPNS) and all-trans-retineic acid(ATRA) by coloning the Nurrl gene and transfecting it into MSCs. Methods Expressing plasmids pcDNA3.1-hygro-Nurrl were cloned,then transfected into MSCs with lipofectamine 2000.To begin with,MSCs were subcultured into 6-wells cultured plate at about 5?10~5 cells/well density and the wells were divided into four groups randomly which were Nurrl+tPNS/ATRA group,tPNS/ATRA group,Nurrl group and control group.Secondly,the plasmids were introduced to the MSCs in Nurrl+tPNS/ATRA group and Nurrl group,then protein expression of Nurrl was identified with immunocytochemistry.Thirdly,after the MSCs and plasmids had been co-cultured for 48 hours,cells in Nurrl+tPNS/ATRA group and tPNS/ATRA group were induced with BME in advance then with tPNS/ATRA in due form.For cells in Nurrl and control group,the only difference was that tPNS/ATRA was replaced with the culture.Finally we compared the different percentage of positive cells in four groups with TH,AChE and GABA antibodies by immunocytochemistry method. Results The immunocytochemical test showed that the MSCs transfected with Nurrl gene expressed Nurrl protein.The percentage of positive cells of TH antibody in Nurrl+tPNS/ATRA group was(38.4?4.6)% distinctly higher than that of tPNS/ATRA group,which was(5.9?3.4)%.Conclusion With tPNS/ATRA induced and immunocytochemistry of TH,positive cells percentage in Nurrl+tPNS/ATRA group was higher than that in tPNS/ATRA group,which showed a statistic difference.And the inducing function of tPNS and ATRA in MSCs differentiating into neurons was definite.