Effects Of Bone Marrow-derived Mesenchymal Stem Cells On The Axonal Outgrowth And Expression Of AKT/GSK-3β/CRMP-2in Rat Brain After Cerebral Ischemia

Object: It has been generally accepted that Bone mesenchymal stem cells (BMSCs)can improve neurological deficit recovery following stroke. The aim of the research is toinvestigate the effects of BMSCs on the axonal outgrowth and expression ofAKT/GSK-3β/CRMP-2signaling pathway in rat cortical neurons followed MCAO. Wesought to potential mechanisms that BMSCs effect on axonal outgrowth in non-contactsystem.Methods:1. BMSCs from SD rats were cultured and identified in vitro.2. The SPF7to9weeks male SD rats were randomly divided into four groups:sham group, MCAO group, MCAO+BMSCs group and MCAO+BMSCs+LY29004group(12rats in each).3. The acute transient focal cerebral ischemia-reperfusion model was established bymodified thread-occlusion method. The sham group underwent the same procedurewithout inserting the suture. LY294002was injected into the lateral ventricle of rats1hour before MCAO. BMSCs (1×106in1ml PBS) were injected into the tail vein of rats24h after MCAO.4. Modified neurological severity score(mNSS) and modified adhesive removal testwere performed before and at1,3,7,10and14days after MCAO to understand thechanges of nerve function in each group rats.5. Rats were sacrificed and the brains taken out14days after MCAO. Proteinexpression level of AKT、pAKT、GSK-3β、pGSK-3β、CRMP-2、pCRMP-2and GAP-43were detected by Western-bolt.Results:1.The BMSCs cultured in vitro were in fusiform shape, arranged in cluster or swirling, and can be successfully induced into osteoblast and adipocytes.2. BMSCs transplantation may improve neural functional in rats after cerebral middleartery ischemia.3. BMSCs transplantation increased the expression of pAKT/AKT、pGSK-3β/GSK-3βand GAP-43, and reduced the level of pCRMP-2/CRMP-2.Conclusion: BMSCs might promote neurological deficit recovery following stroke,which may be related to activation of AKT/GSK-3β/CRMP-2signaling pathway.