Effects Of Adipose-derived Stem Cells Transplantation On The Neuronal Apoptosis And The Expression Of Bcl-2 And Caspase-12 In The Brain Post Focal Cerebral Ischemia In Rats

Object To investigate the effects of adipose-derived stem cells (ADSCs) transplantation on neuronal apoptosis and the expression of bcl-2 and caspase-12 in the brain post focal cerebral ischemia in rats.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 DAPI 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:Heart perfusion,draw the brain from rat,slice on D4,D7 and D14 after MCAO ; Terminal deoxynucleotidyltransferase—mediated biotinylated UTP nick end labeling(TUNEL)staining was used to detect in cell apoptosis;6.3 The expression of bcl-2 and caspase-12 was detected in ischemic region by RT-PCR and immunohistochemistry ;The survive of DAPI -ADSC which were transplanted into MCAO rats was observed by using fluorescence microscopy.7. Statistical analysis.Results 1. The third-generation cells cultured highly expressed CD90 and low expressed CD106and CD34.2. DAPI staining positive cells were observed around the cerebral infarcted area in the ADSC group.3. TUNEL staining revealed that a lot of cell apoptosis was detected in ischemic region post focal cerebral ischemia. TUNEL - positive cells in ischemic region of MCAO +ADSC-treated group were fewer than that in MCAO group and vehicle group.4. RT-PCR and immunohistochemistry revealed that The expression of bcl-2 in the brain tissues of MCAO +ADSC-treated group was higher than that in MCAO grup and vehicle group on D4, D7 and D14 post MCAO (P < 0. 05). The expression of caspase-12 in the brain tissues of MCAO +ADSC-treated group was lower than that in MCAO group and vehicle group on D4, D7 and D14 (P < 0. 05)Conclusions: The transplantation of ADSCs can promote bcl-2 expression and inhibit caspase-12 expression of rats with cerebral ischemia injury and accordingly neuronal apoptosis will be reduced.