Mechanism For The Outgrowth Of Neuronal Axons On Adipose-derived Stem Cell Transplanting For Treatment Of Cerebral Infarction In Rats

Object:To investigate the effects of adipose-derived stem cell (ADSC) transplanting on the outgrowth of Neuronal Axons and the expressions of GFAP, Neurocan, Neuritin, NF-200 in the brain post focal cerebral ischemia in rats and their molecular mechanisms.Methods:1. Adipose-derived stem cells (ADSCs) from rats were cultured in vitro, their specific cellular surface markers were analysed with Immunofluorescence.2. Experimental animal and groups: 54male adult Sprague-Dawly rats were randomly divided into 3 groups: sham-operated group, middle cerebral artery occlusion (MCAO) group and MCAO+ADSC-treated group (n=18).3. A permenant focal cerebral ischemia model was established with the modified Longa's method.4. ADSCs were prelabeled with DAPI in vitro before transplantation.5. Transplantation of ADSC: One day after right MCAO, 30μL of cell suspension containing 1×106 cells were injected into the lateral ventricle of MCAO+ADSC-treated group.6. Before sacrificed at 7d, 14d and 28d after MCAO, the neurological functions were tested by mNSS.7. Materials and Estimates7.1 Heart perfusion, draw the brain from rat, sliced on 7d, 14d and 28d after MCAO.7.2 The survival of DAPI-ADSCs which were transplanted into MCAO rats was observed by using confocal microscopy.7.3 The expressions of GFAP, Neurocan, Neuritin and NF-200 were detected in ischemic region by Western blot.7.4 The expressions of GFAP, Neurocan, Neuritin and NF-200 were observed in ischemic region by Immunofluorescence.7.5 Pathological tissues were observed in ischemic region of different time by H-E.8. Statistical analysisResults1. DAPI staining positive cells were observed around the cerebral infarcted area in the ADSC group.2. ADSCs Transplanting can relive the degree of the neurological functions damage.3. In the brain of MCAO rats,a sheet of dead nerve cells in the central area and lots of Spongiocytes or gliar scard in the peripheral area were observed in different time by H-E.4. Western blot analysis revealed that the expressions of GFAP, Neurocan were lower, Neuritin, NF-200 were higher than that of the MCAO group at 7d, 14d and 28d (P<0.05).Conclusions: The transplantation of ADSC can induce regeneration and repairment of impaired neuronal axons in rat brain after cerebral ischemia, partly by inhibiting the expressions of GFAP, Neurocan and enhancing the expressions of Neuritin, NF-200 in the brain.