| Background: Parkinson's disease is one of the neurological diseases that commonly occurs in the middle-aged and elderly people.The main pathogenesis of the disease is degeneration or necrosis of dopaminergic neurons in the substantia nigra region of the midbrain,which results in the decrease of dopaminergic neurons.Now,some Parkinson's disease patients experimental studys have been carried out.But the domestic study of Parkinson disease mainly stay in the stage for researching animal models of Parkinson's disease.To establish the stable and reliable animal model of Parkinson's disease is the foundation of research on Parkinson's disease.Human is working to find effective ways to treatment Parkinson's disease.Now,drug treatment is the main way to therapeutic the disease.Levodopa is representative drug,followed by surgery and biological agents.With the development of stem cell research in the field of neuroscience,stem cell transplantation has begun to be applied to the treatment of Parkinson's disease,bringing new light to the treatment of Parkinson's disease.In recent years,studies on stem cell transplantation for Parkinson's disease have been carried out in succession,Seeking a relatively effective treatment approach becomes the focus of research.Based on the above background,this study respectively attempted to transplant rat neural stem cells and induced pluripotent stem cells to treat Parkinson's disease rat model,to explore the survival and differentiation of transplanted cells,to evaluate the effectiveness of transplantation through comparison,to explore effective path for cell transplantation treatment of Parkinson's disease.Objective:(1)Establishment of a relatively ideal animal model of Parkinson's disease.(2)To master cell transplantation technology and explore the effectiveness and feasibility of cell transplantation for treating Parkinson's disease rats.Methods: In this experiment,adult male SD rats were used as experimental animals to prepare unilateral lesions of Parkinson's disease(PD)rat models with a stereotaxic apparatus by using 6-OHDA unilateral two-point injection of right medial forebrain bundles.The behavioral of PD rats were identified by apomorphine(APO)-induced rotation test.The behavior of rats was detected by open field test and terrace water maze test.Rat neural stem cells(NSCs)and induced pluripotent stem cells(iPSCs)were cultured to observe cell growth,differentiation and proliferation,and cell characteristics were identified and tested.The cells with better growth status were collected for transplantation,and the transplantation effects were evaluated from the ethological and pathological aspects after transplantation.Result:(1)Successful establishment of a single-sided two-point lesion Parkinson's disease(PD)rat model,the modeling success rate of up to 60%,within two months after surgery,the number of successful models continued to increase,the success rate increased gradually.The unilateral two-point injection of 6-hydroxydopamine(6-OHDA)into the medial forebrain bundle has the advantages of good stability and high success rate.(2)The results of in vitro culture of neural stem cells showed that the cells are suspended in a spherical shape,and the cells gradually aggregate and increase,forming neurospheres,followed by gradual apoptosis.In the case of good cell growth,TH identified as positive.(3)The results of rat Induced pluripotent stem cells culture showed that iPSCs began to change fluids on the fourth day after resuscitation and had the best growth state around the fifth day,and the surface antigens and marker genes were positive.(4)The behavioral results showed that compared with the normal group,the rats in the model group and the cell transplantation group had significant differences in behavioral performance in the open field test and water maze test(P<0.01);Compared with the model group,NSCs transplantation group rats had significant differences in behavioral performance in the open field test and water maze test(P<0.01).Compared with the model group,iPSCs transplantation group rats had differences in behavioral performance(P<0.05),but the improvement effect was not as good as NSCs transplantation group.(5)The results of pathological examination showed that compared with the model group,the positive expression of TH in the right substantia nigra of the NSCs transplantation group rat increased at the third week after cell transplantation,and the morphology of some neurons gradually returned to normal,and the number of neurons increased significantly.Compared with the normal group,the positive expression of TH in the right substantia nigra of the NSCs transplantation group and model group rats significantly reduced,and the neuronal cell morphology changed,the number of neurons significantly reduced.(6)The results of Western Blot showed that Compared with the first week after cell transplantation,the TH protein concentration of NSCs transplantation group rat showed an increasing trend in the third week,and the relative expression of protein increased significantly,the difference was significant(P<0.01).Conclusion:(1)In this study,two-point injecting 6-hydroxydopamine into the right medial forebrain bundle to build a lateral damaging PD rat model was a highly efficient method.The PD rat model was stable and was suitable for stem cell transplantation to treat Parkinson's disease.(2)NSCs transplantation therapy for Parkinson's disease rat models had a good improvement effect within three weeks after cell transplantation;iPSCs transplantation in the treatment of Parkinson's disease rats had a certain improvement effect in the three weeks after cell transplantation,but the improvement effect was not as good as NSCs transplantation group,and there were certain differences between different individuals. |
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