Therapeutic Effect Of Bone Marrow Mesenchymal Stem Cells On Ischemic Stroke

Objective To explore the therapeutic effect and mechanism of bone marrow mesenchymal stem cells(BMSCs)on ischemic stroke in rats.Method (1)The BMSCs were isolated by whole bone marrow adherent method.The proliferation of BMSCs was detected by CCK-8 method and identification of BMSCs by flow cytometry.(2)A rat model of middle cerebral artery occlusion(MCAO)was induced via the modified Longa method.After 24 hours,the rats were blindly divided into sham group,MCAO group,and BMSC group,and BMSCs were injected into the tail vein.(3)The m NSS score,TTC staining and brain water content were measured at 3,7 and 14 days after transplantation.(4)On the 7th day after transplantation,the brains of rats were perfused to prepare frozen sections of brain tissue,then the morphology of cells was observed by HE staining;Detection of GFAP positive cells in and around the infarcted area by immunohistochemical method.(5)The expression level of IL-10 in brain tissue of each group was detected by ELISA method.(6)Immunofluorescence and Western blot were used to detect the positive expression and expression levels of Caspase-3,MAP-2,GAP-43 and GFAP in the infarct area and surrounding areas of the brain tissues of each group.Result (1)BMSCs were successfully isolated,and their growth showed an "S" shape;Flow cytometry detected that BMSCs expressed low CD45 and high expression of CD90 and CD29;(2)The m NSS score,infarct volume percentage and brain water content of rats in the BMSC group were lower than those in the MCAO group at 3,7and 14 days after transplantation;(3)HE staining showed that the infarcted area of the affected side of the brain tissue in the MCAO group was destroyed and dissolved,without obvious contours,loosely arranged,and severe neuronal degeneration.The infarcted area of the BMSC group was relatively intact and the neuronal degeneration was reduced.(4)Immunohistochemical staining showed that the number of GFAP positive cells in BMSC group was significantly higher than that in MCAO model group.(5)on the 3rd and 7th day after transplantation,the expression of IL-10 in MCAO group was up-regulated compared with sham operation group,and IL-10 was also up-regulated in BMSC group compared with MCAO group,but there was no difference between 14 days after transplantation.(6)The results ofimmunofluorescence staining and Western Blot showed that,compared with the sham group,the expression of GFAP,GAP-43 and Caspase-3 was increased,and the expression of MAP-2 was decreasedin the MCAO model group,compared with the MCAO model group,BMSC In group Caspase-3 decreased significantly,and the expression of MAP-2,GFAP and GAP-43 increased significantly.Compared with the MCAO model group,Caspase-3 was decreased,and the expression of MAP-2,GFAP and GAP-43 was increased in the BMSC group.Conclusion BMSCs transplantation can promote the recovery of neurological function and reduce the volume of cerebral infarction in MCAO model rats,possibly through mechanisms such as nerve regeneration cell proliferation,inhibition of cell apoptosis,inflammatory reaction and axon regeneration.