The Study Of The BFGF Expression After Grafting Bone Marrow Stromal Cells Associated With Bcl-2 Gene For Treating Cerebral Ischemia In Rats

PrefaceThe disease incidence rate and the case fatality rate of the ischemic cerebrovasculardisease increase year by year, The recovery after neurous damage is still a difficultproblem. Bone marrow stromal cell (BMSCs) are the stem cells with multilineagedifferentiation potential and self-replication abilitie, so become the good method incerebrovascular disease. But the apoptosis of BMSCs is a problem. Bcl-2 gene is anidentified anti-apoptosis gene. This research coadministration BMSCs with the Bcl-2gene, observing the apoptosis of the cells,modified Neurological Severity Scores,whether the expression of basic fibroblast growth factor (bFGF) can be improved andwhether coadministration of BMSCs with bcl-2 into the MCAo rats improves functionalrecovery after stroke.MethodsForty adult rats were used to establish ischemia-reperfusion rat models bytransient occlusion of the left middle cerebral artery (MCAo),and to induce focalcerebral ischemia for 2h, followed by reperfusion, and were assigned randomly into 4groups: the control group (n=10),the pure BMSCs group (n=10),the pure bcl-2 genegroup(n=10)and the experimental group (BMSCs with bcl-2 group)(n=10). Three hourslater, pLXSN -bcl-2 was injected into through interial icarortid artery. The BrdU-markedBMSCs were injected into through tail vein after 24 hours reperfusion. Each group ofrats were divided into 3d and 14d two time spots(each time spots 5 rats). NeurologicalDeficits Scores(NDS) were observed at 3d,14d. Their cerebral tissues were obtained at 3 or 14 day after 2 hours of MCAo. Brain sections were stained with hematoxylin andeosin (HE) for surveying the volume of infarction. Double-stainingimmunohistochemistry of brain sections was used to identify bFGF,bcl-2 proteinexpression level and distribution and quantity in the brain of the BrdU-marked BMSCs.Date were acquired from different fields in the margin of the infarctionfocus, hippocampus and corpus striatum region by analysing the positive areapercentage or counting the number of positive cells at high powermagnification(400X). SPSS 13.0 statistical soft was used to analyse the date. Alldates were expressed as x±s. A value of (p＜0.05) was considered statisticallysignificant.Results1.The score of neurologic impairment in the experimental group at3d(5.68±0.25)was greatly lower than that in the physiological saline group, and that at14d (2.06±0.47)was greatly lower than that in the control group(p＜0.05).2.The number of cell apoptosis significantly decreased in the experimentalgroup.The cell apoptosis neurous in pure BMSCS group or pure bcl-2 group was greatlydecreased compared with the control group.3. Many of the BrdU-labeded BMSCs could be seen at the ischemic hemisphere inthe experimental group and pure BMSCs group. The number of engrafted BMSCs in theexperimental group were significantly higher than that in the pure BMSCs group.4.The expression of bFGF were significantly increased in experimental group thanthat of the other groups(p＜0.05), The expression of bFGF in BMSCS group or bcl-2group was greatly increased compared with the control group.5.The expression of bcl-2 at 3d,14d significantly increased in experimental groupcompared with the control group.ConclusionThese result suggest that both BMSCs and bcl-2 gene can therapy cerebralischemic, coadministration of bone marrow stromal cells (BMSCs) with bcl-2 couldenhance the expression of bFGF, improve the nerological function combingtreatment could provide a potentiol therapertic strategies to the treatment of ischemic stroke.