Researches About The Mechanisms Of SDF-1/CXCR4Signal Transduction Pathway In The Treatment Of Parkinson's Disease By Neural Stem Cell Transplantation

Objective(1) To research the effects of neural stem cell transplantation in the treatment of Parkinson's disease.(2) To explore the molecular mechanisms of neural stem cell transplantation in the treatment of Parkinson's disease by SDF-1/CXCR4signal transduction pathway.Methods(1) To dissect the neural stem cells in the ventri-mesencephalon of E14d rats embryo and to culture those cells in the neural stem cell culture medium without blood serum. And then, we observed the proliferation and differentiation of those neural stem cells. The stem cell characteristics and differentiation potency of those cultured cells were identified by immunocytochemistry.(2) To establish the PD models of rats by injecting6-OHDA into the substantia nigra and medial forebrain bundle of rats through stereotaxis.(3) The PD model rats were randomly allocated into four groups:Control group(without the transplantation of neural stem cells and injection of AMD3100), Neural stem cell transplantation group, Neural stem cell transplantation plus injection of AMD3100group, Injection of AMD3100group.(4) To inject well-growing neural stem cells cultured for3nd generation into the substantia nigra and medial forebrain bundle of rats each in Neural stem cell transplantation group and Neural stem cell transplantation plus injection of AMD3100group.(5) The rats in Neural stem cell transplantation plus injection of AMD3100group and Injection of AMD3100group were subjected to injection of AMD3100(1.25mg/kg/d), a CXCR4antagonist, into the abdominal cavity on each day after neural stem cell transplantation.(6) To induce the rats'rotation by injecting APO(lml,0.2mg/ml) into the abdominal cavity each on1W,2W,3W,4W after neural stem cell transplantation and behavior scores were assessed while the occurrence of rotation after injection of APO into the abdominal cavity each on1W,2W,3W,4W.(7) The rats in each groups were killed on1W,2W,4W after neural stem cell transplantation. The total RNA of brain tissue in affected side was extracted by TRIzol(Invitrogen). Then the gene expressions of SDF-1and CXCR4were detected by RT-PCR.(8) The rats in each groups were killed on1W,2W,4W after neural stem cell transplantation. The protein expressions of SDF-1and CXCR4in the brain tissue in affected side were detected by Western Blot.(9) All the measurement data were analyzed by SPSS17.0and were represented by×±s, P<0.05indicated that the discrepancy had statistical significance.Results(1) The neural stem cells from the ventri-mesencephalon of E14d rats represented the potency of proliferation and formation of neurospheres. The neural stem cells cultured for6days in the isolation medium were Nestin positive by immunocytochemistry. Special molecule maker of dopamine neuron in the neural stem cells after differentiation were expressed by RT-PCR and TH immunocytochemistry.(2) We established the PD models of rats successfully by injecting6-OHDA into the abdominal cavity and these models were stable. The PD models didn't recover by behavior scores.(3) The APO-induced rotation on1W,2W,3W,4W and behavior scores of the rats in Neural stem cell plantation group were improved than those in Control group. The APO-induced rotation on1W,2W,3W,4W and behavior scores of the rats in Injection of AMD3100group and Neural stem cell transplantation plus injection of AMD3100group had no significant diversity in contrast to those in Control group.(4) The gene expressions of SDF-1and CXCR4in Neural stem cell transplantation group were increased than those in other groups. The protein expressions of SDF-1and CXCR4in Neural stem cell group were also increased than those in other groups. The trend of protein expressions was accordiance with RT-PCR. So, we considered that the gene expressions were up to the protein expressions of SDF-1and CXCR4.Conclusion(1) The neural stem cell transplantation could improve the movement disorders of PD rats.(2) The neural stem cell transplantation was effective in the treatment of PD and the mechanisms were associated with the increase of SDF-1expressions and SDF-1/CXCR4signal transduction pathway.