| Object： To investigate the effects of treadmill exercise and Adipose-derived Stem Cellstransplantation on the neurologic function and the expression of axonal growth inhibitingfactor Nogo A and its receptors NgR、PirB in rat brain after cerebral ischemia.Methods：1. Adipose-derived stem cells (ADSCs) from rats were cultured in vitro.2.108male adult Sprague-dawley rats were randomly divided into five groups:sham-operated group、MCAO group、transplantation group、exercise group and combinationgroup. The later4groups were randomly divided into2subgroups according to time pointsafter MCAO:7and28days,12rats in every subgroup. Every rat was placed on the motordriven treadmill and trained to run over a three-day adaptation period.3. A MCAO model was established with the modified Zea-Longa’s method.4. At24h after post-MCAO,1×106ADSCs were injected into the lateral ventricle of rats intransplantation group and combination group.5. From48hours after MCAO, rats in exercise group and combination group were forced torun on the treadmill for30min per day until the time set before. Rats in other groups were lefton the treadmill for30mins and were not made to run.6. Rats in every group underwent behavioral functional tests prior to the MCAO procedureand at4,7,14and28days after MCAO, and the modified neurologic severity scores (mNSS)was used to measure various aspects of neurologic function of rats.7. At7days after MCAO，hematoxylin-eosin staining(HE staining) was used to determinehistopathological damage in rat brain after cerebral ischemia. At28days after MCAO，TTCstaining was used to determine the volume of infarct of brain in each group.8.At7and28days after MCAO，the tissue corresponding to the peri-infarct cortex wasdissected for expression level of Nogo A and its receptors NgR、PirB by real-time PCR andWestern blot, and localization of Nogo A and its receptors NgR、PirB by immunofluorescentstaining. Results：1. The neurological functions were gradually better in each group. Intransplantation group、exercise group at14d and28d after MCAO were better than that inMCAO group (P<0.05), combination group was better than that in MCAO group(P<0.01orP<0.05); At7d after MCAO, HE staining showed injured cells with twisted axonal processesand cell loss were evident throughout ischemic cortex, and those changes were very few ineach treatment group; At28d after MCAO, TTC staining showed that the volume of infarct ofbrain was smaller in combination group than the othe groups(P<0.01or P<0.05).2. Realtime-PCR and Western blot analysis revealed that the expression of Nogo A、NgR andPirB in combination group was significantly downregulated at28d after post-MCAOcompared with transplantation group and exercise group,(P<0.01or P<0.05).3. Immunofluorescent staining analysis showed that Nogo A was mainly expressed inoligodendrocytes in peri-ischemic brain area and little expressed in neurons；NgR, however,was expressed mostly in neuronal perikarya, and seldom in neuronal processes inperi-ischemic brain area, while PirB was mostly expressed in neural processes inperi-ischemic brain area.Conclusion： ADSCs transplanting and treadmill training can improve the neurologicalfunctions of rats and have a synergistic effect. It can down regulate the expression of Nogo Aprotein and its receptors NgR、PirB in ischemic brain after MCAO in rats．... |
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