NgR Gene Silencing Combined With NT-3Gene Over-expressing On The Effects Of The Proliferation Of Rat Neural Stem Cells

Objective:With the development of transportation and construction business,the incidence of spinal cord injury showed a rising trend year by year. The treatment of spinal cord injury is a worldwide difficult problem. Neural stem cells transplantation and gene therapy are the most important approaches to novel strategies for spinal cord injury.The research of NgR gene silencing combined with NT-3gene over-expressing on the effects of the proliferation of rat neural stem cells provide the necessary experimental basis for research and clinical applications of in vivo transplantation.Methods:According to the literature, select the highest silencing efficiency siRNA to construct NgR interference vector, at the same time build NT-3over-expressing vector plasmid. Package the slow virus each other;Prepare the neural stem cells; Slow virus transfection of neural stem cells, this step is divided into five groups:Blank control group (Con), The slow virus empty vector group (NC), NT-3over-expressing group, NgR interference group, NT-3over-expressing+NgR interference group; Using Western Blot method for the detection of target gene expression for NT-3over-expressing group and NgR interference group; Collect after infection of each neural stem cells for MTT detects the proliferation ability of cells; PI single dye flow cytometry.Results:1. MTT:Inoculating plates within72hours the cells OD all showed an upward trend at different time points. This shows that cell proliferation mitotic ability have varying degrees of increase, in anti-NgR+NT-3group of neural stem cells enhance the most obvious (p<0.05), while in NT-3group and anti-NgR group of proliferation of neural stem cell division ability is also strong in the blank control group and slow virus empty vector group (P<0.05).2. PI single dye flow cytometry:In S, group anti-NgR+NT-3neural stem cells make up36.88%of the set of all cells, higher than the other groups. In S, group NT-3neural stem cells make up21.17%of the set of all cells. In S, group anti-NgR neural stem cells make up20.73%of the set of all cells, both higher than blank control group and slow virus empty vector group. It shows that group anti-NgR+NT-3neural stem cell proliferation mitotic ability was stronger than the other groups (P<0.05), While in group NT-3and group anti-NgR neural stem cell proliferation mitotic ability were significantly stronger in the blank control group and slow virus empty vector group (P<0.05).Conclusions:1. NgR gene silencing and NT-3gene over-expressing make the proliferation of neural stem cell division ability increase apparently by oneself.2. NgR gene silencing combined with the NT-3gene over-expressing can make the proliferation of neural stem cell division ability increase apparently, stronger than the NgR gene silencing and NT-3gene over-expressing effect of neural stem cells.