Mechanism On Adipose-derived Stem Cell Transplanting For Treatment Of Focal Cerebral Infarction In Rats

Object To investigate the effects of adipose-derived stem cell (ADSC) transplanting on the angiogenesis and the expression of bFGF,TGF-β1,VEGF in the brain post focal cerebral ischemia in rats and its molecular mechanism.Methods 1. Adipose-derived stem cell (ADSC) from rats were cultured in vitro,their specific cellular surface markers were analysed with flow cytometer.2. Experimental animal and groups:72 male adult Sprague-Dawly rats were randomly divided into 4 groups:sham-operated group,middle cerebral artery occlusion (MCAO) group,vehicle group and MCAO+ADSC-treated group (n=18).3. A permenant focal cerebral ischemia model was established with the modified Longa's method.4. ADSC were prelabeled with CFSE in vitro before transplantation.5. Transplantation of ADSC:One day after right MCAO,30μL of cell suspension containing 1×106 cells were injected into the lateral ventricle of MCAO+ADSC-treated group and the same dose of PBS was given to the vehicle group.6. Materials and Estimates.6.1 Heart perfusion,draw the brain from rat,slice on D4,D7 and D14 after MCAO.6.2 Immunohistochemistry identification of VIII-Ag expression.6.3 The expression of bFGF,TGF-β1 and VEGF was detected in ischemic region by RT-PCR and immunohistochemistry .6.4 The survive of CFSE-ADSC which were transplanted into MCAO rats was observed by using confocal microscopy.7. Statistical analysis.Results 1. CFSE staining positive cells were observed around the cerebral infarcted area in the ADSC group.2. Immunohistochemistry revealed that a lot of microvessel proliferated in the injured cortex. The microvessel density in the brain tissues of rats treated with ADSC was higher than that in MCAO group and vehicle group (P<0.01).3. RT-PCR and immunohistochemistry revealed that the expression of bFGF,TGF-β1 and VEGF were higher than that of the MCAO group and vehicle group on D4,D7 and D14 (P<0.01).4. CFSE-ADSC which were transplanted into MCAO rats could survive in the brain post focal cerebral ischemia by using confocal microscopy ,showed the ability of secreting bFGF,TGF-β1 and VEGF.Conclusions: The transplantation of ADSC can lighten the neurons injury in the brain post focal cerebral ischemia in rats partly by promoting the revascularization of cerebral ischemia and enhancing bFGF,TGF-β1 and VEGF synthesis in brain.