The Research Of SDF-1/CXCR4Pathway Molecular Mechanism Of Endogenous Neural Stem Cell Transplantation For Treatment Of Parkinson’s Disease

Objective(1) Isolate and culture neural stem cell, then translate it to the corpus stratum of Parkinson’s disease model mouse. Explore the therapeutic effect of endogenous neural stem cells for Parkinson.(2) With SDF-1/CXCR4signaling axis as the access point, explore the endogenous neural stem cell and illustrate its function.Provide a new idea of cell transplantation therapy for PD in the future.Method(1) Isolate the ventral brain tissue of rat embryo of E14d,and made it into NSCS single cell suspension by mechanical percussion. Culture it in serum-free medium to observe its proliferation and differentiation. Use immunohistochemisty to identify the characteristics and differentiated potential of cultured stem cells.(2) Inject moderately6-hydroxydopamine (6-OHDA) with Stereotaxic method to the site of the medial forebrain of the healthy adult rat,Created a rat model of Parkinson’s disease.(3) After two weeks, abdominal inject APO to rat. The APO can induce rat to rotate, and Choose those animals who greater than210transfer/30min (7rpm) for the success model of PD rat. The PD rat was divided randomly into four groups: Group A:neural stem cell transplantation group; Group B:neural stem cell transplantation+AMD3100group; Group C:AMD3100group; Group D:control group (no transplantation of neural stem cells, no injection of AMD3100);(4) After primary cultured neural stem cells in culture by two passages, Choose the neural stem cells of good growth state, and transplant it into the striatum of PD rats.(5) After transplanted, group B and C group was abdominal injected with SDF-1/CXCR4blocking agent AMD3100,/(6)Observe behavioral change of neural stem cell transplantation for PD rats.,.(7)After cell transplantation,1week,2weeks and4weeks,partially executed each experimental group rats and detected Nestin, SDF-1and CXCR4in rats Tissue with immunity fluorescence method.(8) Statistical methods:this experiment measuring data use all mean±standard deviation (plus or minus S) to illustrate, and using SPSS17.0statistical software, with P=0.05for inspection standards for variance analysis.Result(1) Cells isolated from E14fetal rat brain ventral parts have ability of proliferation and to form neurospheres. Neurospheres express Nestin, RT-PCR and immunohistochemistry results showed that most cells express the dopamine neuron-specific molecular markers.(2) The way of unilateral injection of6-OHDA that prepare the side of the striatum pathway damage PD animal models was success and has a fairly long period of stability. Behavioral testing did not appear the phenomenon of self-recovery, As a variety of methods of treatment effect in animal models of PD.(3) PD rat model of neural stem cell transplantation group, After1W,2W,3W and4W, APO induced rotational behavior in the early model was successful and the control group decreased significantly.AMD3100injection group and the neural stem cell transplantation+AMD3100injection group,after1W,2W,3W and4W,.APO induced rotation behavior show that Compared with the control group did not change significantly,And with the same control group for6weeks showed a rising trend.(4)Successly detected Nestin, SDF-1and CXCR4expression of rats’ brain tissue with immunity fluorescence method..there is an obvious difference in neural stem cell transplantation group between the other group and the control group. There is obvious migration tendency in the neural stem cells transplantation group rats that endogenous neural stem cells from the SVZ area into the striatum area and is consistent with expression trend of SDF-1and CXCR4.Conclusion(1) The isolated cells of E14fetal rat brain ventral parts have a strong proliferation and the formation of the ability of dopaminergic neurons, is an ideal source of stem cells to treat Parkinson’s disease.(2)6-OHDA into the stereotactic intracranial injection prepared by PD rat model has good stability and repeatability.(3) exogenous neural stem cells transplantation is effective in the treatment of Parkinson’s disease,Is likely to achieve value-added and migration of endogenous neural stem cells within the activation Endogenous neural stem cells of value-added migration and SDF-1/CXCR4axis is closely related, and can have been injected with the amount of AMD3100effectively prevent.