Comparison Of Intravenous And Intracarotid Route For Bonemarrow Derived Mesenchymal Stem Cells In The Treatment Of Acute Cerebral Infarction In Model

Objective:To investigate the difference of GFAP、VEGF、Bcl-2、Bax and TUNEL positive cells in the ischemia penumbra, the level of TNF- in serum, the score of mNSS, the count of body weight and the distribution of stem cells in organs between intravenous and intracarotid route for bone marrow derived mesenchymal stem cells in the treatment of MC AO 6 hours. To investigate the difference of intravenous and intracarotid delivery route and the mechanism of BMSCs in the treatment of MCAO Methods:1. The middle cerebral artery occlusion model was conducted by modified intraluminal suture method.2. The bone marrow derived mesenchymal stem cells were isolated, purified and cultured by whole bone marrow adherent culture and identified by flow cytometry. BMSCs were dyed with CM-dil for tracing.3. Transplantion was operated at MC AO 6 hours by intravenous and intracarotid route respectively.4. 7days after transplantation, the score of mNSS and the count of body weight were evaluated.5. The level of TNF-α in serum was detected by ELISA.6. The expression of GFAP 、 VEGF 、 Bcl-2 and Bax were detected by immunohistochemistry.7. Cell apoptosis was detected by TUNEL staining. Results:1. The expression of surface antigen CD90、CD29、CD106、CD11b、CD34、CD45 were respectively 95.7%、97.3%、52.7%、6.01%、2.95%、2.26%.2. Compared with PBS group, the scores of mNSS was significantly decreased in the BMSCs groups, the difference between intracarotid and intravenous BMSCs group was not showed. Compared with intracarotid PBS group, the count of body weight was increased in the intracarotid BMSCs group, and the result was the same between the intravenous PBS and BMSCs group. Compared with intravenous PBS group,the difference was not show in the intracarotid BMSCs group. The difference between intracarotid and intravenous BMSCs group was not showed.3. The GFAP positive cells were observed in the ischemia penumbra surrounding the ischemia core mainly. Regardless the delivery route, compared with PBS group, the number of GFAP positive cells was significantly increased in the BMSCs groups, the difference between intracarotid and intravenous BMSCs group was not showed. The VEGF positive cells in the ischemia penumbra were observed, Compared with intravenous PBS group, the mean optical density(MOD) was significantly increased in the intravenous BMSCs groups, compared with intracarotid PBS group, the MOD was not increased significantly in the intracarotid BMSCs group. The difference between intracarotid and intravenous BMSCs group and was not showed.4. Compared with intravenous PBS group, the mean optical destiny(MOD) of VEGF was significantly increased in the intravenous BMSCs groups, the mean optical destiny(MOD) of VEGF in the intracarotid BMSCs group was increased but the difference was not showed compared with the intracarotid PBS group, the difference between intracarotid and intravenous BMSCs group was not showed.5. The expression of bcl-2 was observed ubiquitously throughout the brain tissue, heavy expression was shown in the ischemic penumbra. Compared with PBS groups, the mean optical destiny(MOD) of Bcl-2 positive cells was significantly increased in the BMSCs group, the difference between intracarotid and intravenous BMSCs group was not showed. The expression of Bax was observed in the ischemia penumbra mainly. The MO D of Bax positive cells was not significantly different among all groups.6. The level of TNF-αin serum was significantly decreased in the BMSCs groups, when compared with the PBS group. The difference between intracarotid and intravenous group was not showed.7. The apoptosis cells were observed in the ischemia hemisphere only, not in the uninjured hemisphere. Compared with PBS groups, the number of TUN EL positive cells and was significantly decreased in the BMSCs groups. The difference between intracarotid and intravenous BMSCs group was not showed.8. BMSCs dyed with CM-dil were discovered in lung and spleen concentrated, in the ischemia hemisphere only, not in the uninjured hemisphere. The number of BMSCs distributed in the liver and kidney was less than lung and spleen, BMSCs distributed in the heart was not showed. The difference of stem cell’s distribution in organs between intracarotid and intravenous BMSCs group was not showed. Conclusion:BMSCs can promote renew of ischemia reperfusion through enhancing the expression of GFAP and VEGF to maintain the integrity of neurovascular unit, improving the expression of Bcl-2, reducing the apoptosis and TNF- in serum. There is no difference between intravenous and intra-arterial route for bone marrow derived mesenchymal stem cells in the treatment of MCAO 6 hours and in the distribution of BMSCs in the organs, the intravenous route is optimal for tiny invasion and high dose of cells.