Study On BMSCs Transplantation In Pakinson's Disease

Objective:To explore the effect and the mechanism of BMSCs transplantation in Parkinson's disease.At the same time,to evaluate the difference between the two approaches of transplantation.To grope the stratagem of tracking living fluorescent cells by using UVP Imagining system.Methods:PD rat models were established,and randomized into three groups on the basis of BMSCs-transplanted approach:brain-approach group,vein-approach group and controlled group.Based on the adherent characteristic,rat BMSCs were cultured and purified.Then they were marked by GFP through viral vector.The gray scale of GFP-BMSCs imagines made by UVP LS Imageing system was measured;as a result,the regression curve of the GFP-cells' amount and area density was deduced.According to the curve,the amount of living transplanted GFP-BMSCs was calculated at the different intervals after BMSCs transplantation.The effect and mechanism were explored from different aspects.(1) APO-induced rotation was quantified at 2,4 and 8 weeks after BMSCs transplantation to evaluate the therapy effect.(2) The level of DA and its' metabolin,DOPAC and HVA,in different groups was detected by HPLC-ECD to analysis the secret- function of dopaminergic neuron.(3) The cell morphology of transplanted BMSCs was observed,and so did the surrounding tissue.The differentiation of transplanted BMSCs was witnessed by fluorescent immunohistochemistry.Results:Rat BMSCs marked by GFP could be cultured stably,and GFP gene could be well expressed after freezing and thawing.Living fluorescent cells had fine linear correlation with area density.The number of living transplanted GFP-BMSCs in PD rat model was semiquantified,and it degraded along with the interval prolonged after transplantation.APO-induced rotation of PD rat was significantly reduced in brain-approach group and vein-approach group compared with controlled group 2,4 and 8 weeks after BMSCs transplantation(p＜0.05),and so did brain-approach group compared with vein-approach group(p＜0.05).It was also observed that after BMSCs transplanted,APO-induced rotation in brain-approach and vein-approach significantly reduced compared with pre-transplantation(p＜0.05).The level of DA and DOPAC,HVA in brain-approach group and vein-approach group was higher than controlled group(p＜0.05) while brain-approach group was higher than vein-approach group(p＜0.05).No apparent change,such as inflammation or over proliferation,was observed in the surrounding tissue in the three groups.2,4 and 8 weeks after transplantation,GFP-positive cells were found in cerebrum both in brain-approach group and vein-approach group.GFP-positive cells were found in the opposite hemisphere in brain-approach group.4 and 8 weeks after transplantation,the expression of TH-1 in brain-approach group and vein-approach group was higher than controlled group(p＜0.05),particularly brain-approach group.Double positive cells(GFP~+/TH-1~+) were founded in both groups.The expression of GFAP in brain-approach group and vein-approach group was higher than controlled group(p＜0.05),and double positive cells(GFP~+/GFAP~+) were founded in both groups.The expression of NSE in brain-approach group and vein-approach group was higher than controlled group(p＜0.05),particularly brain-approach group.Double positive cells(GFP~+NSE~+) were founded in both groups.Conclusions:(1) BMSCs transplantation is an effective and safe therapy for PD rat models.(2) The mechanism of the therapy includes the substitution of differentiated dopaminergic neurons coming from transplanted BMSCs and nutritional effect of differentiated astrocytes coming from transplanted BMSCs. Furthermore,transplanted BMSCs provide a microenvironment which is suitable for repairing damage.(3) Both the brain-approach and the vein-approach are benefit for PD therapy,but the former is better than the later.(4) UVP LS imaging system can semiquantitative and track fluorescent cells.