| Objectives：We tested the hypothesis that whether Adipose-derived mesenchymalstem cells (ADSCs) transplanted into brain via internal carotid artery after acuteischemic stroke, can survive and migrate to the ischemic boundary zone (IBZ),differentiate into neural cells and improve neurological functional recovery. Meanwhileprobe the potential therapeutic mechanism of ADSCs involving in the cerebral ischemicstroke.Methods: ADSCs were harvested from adult SD rats for cellular identification,and prelabeled with CM-Dil. Rats (n=30) were subjected to transient (1.5h) middlecerebral artery occlusion (MCAo), received grafts at3d and were euthanized at28daysafter MCAo. Test groups consisted of:1) Intracarotid transplantation of ADSCs afterMCAo (n=10);2)Intracarotid transplantation of saline after MCAo (n=10);3) rat afterMCAo without handle(n=10).Immunofluorescence was used toidentifyADSCs’distribution and express in vivo. Histology and Immunohistochemistrywas used to observe anotomy pathology and evaluate the ischemic lesion volume.Behavioral tests (modified neurological severity score mNSS) were performed beforeand after MCAo. TUNEL was used to assess the apoptosis in the IBZ among groups.Results:1. mNSS score: there were no significant differences of behavior testamong all groups before MCAo(P＞0.05)；there were significant differences ofbehavior test at time set points before and after MCAo in all groups(P＜0.01)；multiplecomparison results showed that: there were statistical significance of differences forbehavior test between ADSCs group and saline group，ADSCs group and blank group(P＜0.01)；there were no significant differences between blank group and saline group(P＞0.05)；all rats appeared serious behavior deficit after3d of MCAo,take onprogressive palliation of behavior deficit till28d.there were notably neurofunctionalimprovement after4d of MCAo (P＜0.05).2. infarction volume showed that ADSCs group：35.5%±11.9%；saline group36.9%±9.1%；blank group36.3±9.1%(P＞0.05)3.Histology and IHC results：Under photomicroscope, A nearly complete loss ofparenchymal cell immunoreactivity (NeuN,GFAP) was observed in the ischemic core ofthe striatum and cortex in all MCAo rats. Brain tissue deformations, edema,degeneration, damaged and relatively intact cells were readily observed in the IBZ of allMCAo rats.4. Under laser-scanning confocal microscope, the Numbers(17824±3624) ofCM-labled ADSCs measured from an average of5histology slides (50-um interval) peranimal, ADSCs in the ipsilateral hemisphere (21853±3225) than in the contralateralhemisphere (119±28, p＜0.05)28days after MCAo. Double immunostaining showedthat, from a total of CM-labled cells(22000±663) within5slides per animal,approximately (3.3±1.6%)of CM-labled cells expressed GFAP (astrocytes) and(1.5±0.8%)expressed NeuN (neurons).5. Under fluorescent microscope, ADSCs weredistributed throughout the ischemic regions in the MCA territory. ADSCs were mainlyin the ipsilateral hemisphere at the level of bregma(-1,+1) of the cortices, and therostral-caudal axis of the striatum of the ipsilateral hemisphere. Many more (~90%)CM-labled cells localized in the ischemic core and boundary zone than in the relativelyintact regions of the ipsilateral hemisphere.IHC staining showed that GFAP expressionwas notably lower in ADSCs group than in saline group, suggesting reduced IS-inducedgliosis after ADSCs treatment. We also examined the neuronal apoptosis peri-ischemiaby TUNEL staining，ADSCs treatment remarkably decreased the number of apoptoticcells. IHC staining also revealed a notably increased number of BrdU-positive cells inADSCs group than in saline group。Conclusion:1.ADSCs can survive, migrate to brain ischemic boundery zone(IBZ);2.ADSCs can partly differentiate into phenotypic neuroid cells;3. ADSCs can Significant ameliorate functional recovery after stroke;4. ADSCs Treatment groups attenuated astroglial reactivity, reduces cell apoptosis,and induced endogenous cells proliferation compared with contrast groups;5.The therapeutic mechanism of ADSCs may probably be the interwoved of neuralprotection and neural regeneration.via gliosis inhabitation,antiapoptosis andendogenous proliferation et al. ADSCs may be a roboust means for stroke patients in theclinic. |
PDF Full Text Request