The Expression Of GDNF With Combination Theraphy To Brain Ischemia Using BMSCs And Bcl-2 In Rats

IntroductionThe latest studies show that bone marrow stromal cells (BMSCs) are nonhemopoiesis stem cells except hemopoiesis stems cells in bone marrow induced to differentiation nerve cells and glia cells, so BMSCs transplanted into cerebral infarction promote nerve function recovery. Bcl-2 gene can resist apoptosis, bcl-2 treats ischemia by inhabiting apoptosis, decreasing volume, improving nerve function.Glial cell line-derived nerotrophic factor,GDNF inhabits infarction, lightening hydrocephalus, decreasing apoptosis,avoiding industry of delated nerve cells.The objective of our experiment is whether combination theraphy to brain ischemia using BMSCs and Bcl-2 can not only cooperate or enhance protection but also increse ex-pression of GDNF, So we expect to break a new path for treatment of stroke.Materials and MethodsHealthy Wistar rats used by Longa method (the reformative suture technique) were induced to middle cerebral artery occlusion (MCAO) model, Two hours after the ischemia reperfusion was implemented to those rats, 40 adult rats were assigned randomly into 4 groups,nothing group: 10 rats, Bcl-2 group: 10, BMSCs group: 10, BMSCs+Bcl-2 group:10.Three hours after reperfusion was implemented to those rats,P1XSN-bcl-2 genes were injected into internal carotid artery.24 hours after reperfusion, Bone marrow stromal cells (BMSCs) were injected into trail vein BMSC3×10~6. 3 days and 14 days after each group MCAO Rats were euthanized and targets were detected 3 days (n=5), 14 days (n=5).Bcl-2 protein and GDNF and BMSCs expression were determined through immunohistochemistry. After experimental rats were injected into ahdomen by 10% water match chlorine aldehyde, Then the brains were removed, postfixed, embeddded inparafin, and consecutively sectioned (6um thick coronal sections), immunohistochemistry dyed. The distribution of apoptosis neurons was checked out by TUNEL techniques. All the results were transformed to datas through image analysis system,which were expressed as (?)±s.The values were analyzed by ANOVA and test of SPSS software. A value of P＜0.05 was considered significant,obvious significance was set at P＜0.01.Results1. Bcl-2 protein expression: There was significant increase on the expression of Bcl-2 protein in the control groups (Bcl-2 group,Bcl-2+BMSCs group) compared to nothing group(p＜0.05).2. The expression of BMSCs: There was significant increase on the expression of BMSCs in the Bcl-2+BMSCs group compared to BMSCs group (p＜0.05).3. The apotosis of nerve cells: There was significant decrease on the apotosis of nerve cells in Bcl-2+BMSCs group compared to other groups(p＜0.05).4. The expression of GDNF: There was significant increase on the expression of GDNF in Bcl-2+BMSCs group compared to other groups (p＜0.05).ConclusionCoadministration of bone marrow cells (BMCs) and Bcl-2 into ischemic /refusion of rats after MCAO increase the exprssion of Bcl-2 and GDNF,increase the BMSCs,decrease the apoptosis of nerve cells.