| Objective:To study whether transplantation of extracellular singal-regulated kinase (ERK) gene modifided bone marrow mesenchy-mal stem cells (BMSCs) on cerebral ischemia has better neuroprotection than simple BMSCs transplantation. Methods:ERK1/2recombinant plasmid was constructed and transfected into BMSCs, in order to establish cell line which overexpressed ERK1/2(ERK1/2/BMSCs). Temporary middle cerebral artery occlusion (MCAO) rat models were established by surgical operation and were randomly divided into BV/BMSCs group, ERK1/2/BMSCs group and control group, then transplanted BV/BMSCs, ERK1/2/BMSCs and saline by stereotaxic technique respectively at3days after reperfusion. Neurological Severity Scores(NSS) were performed to test the neurological function change of rats at1week and2weeks after cell transplantation, then rats were sacrificed. Cerebral infarction volume, pathological features, number of neurons around peri-ischemic frontoparietal cortex were evaluated by TTC staining, HE staining and Nissl’s staining respectively. The survival number of implanted BMSCs was observed under fluorescence microscope, immunofluorescent staining was used to observe the expressions of glial fibrillary acidic protein(GFAP) in the periphery of ischemia. Results:1. RT-PCR and western blot test showed that ERK1/2/BMSCs cells overexpressed ERK1/2.2.NSS of ERK1/2/BMSCs group and BV/BMSCs group declined more obviously than that of control group at1week and2weeks(P<0.05), and NSS of ERK1/2/BMSCs group was lower than that of BV/BMSCs group on two timepoint(P<0.01).3. The brain lesion volume of ERK1/2/BMSCs group and BV/BMSCs group decreased more (P<0.05) than that of control group, but the difference between ERK1/2/BMSCs group and BV/BMSCs group is not significant(P>0.05).4. Nissl staining showed that the number of neurons around peri-ischemic frontoparietal cortex of ERK1/2/BMSCs group was obviously more than that of BV/BMSCs group and control group (P<0.05) at first week. At2weeks timepoint the number of neurons of ERKl/2/BMSCs group and BV/BMSCs group was more than that of control group, and that of ERK1/2/BMSCs group is obviously more than BV/BMSCs group(P<0.05).5. The survival number of transplanted cells in ERK1/2/BMSCs group is more than that in BV/BMSCs group(P<0.05) at2weeks, but there was no obvious difference between the two groups at1week.6. The number of GFAP-positive cells from periphery of ischemia of ERK1/2/BMSCs group and BV/BMSCs group were more than that of the control(P<0.05), and that of ERK1/2/BMSCs group was more than that of BV/BMSCs group (P<0.05). Conclusion:1. BMSCs which express stable ERK1/2is constructed successfully. Compared with transplantation of BV/BMSCs, transplantation of ERK1/2gene modified BMSCs is more effective in improving neurological neurological deficit of MACO rats, and promote the recovery of motor and sensory function.2. Transplantation treatment of ERK1/2/BMSCs and BMSCs on ischemic stroke can both reduce the cerebral infarction volume and better the pathological changes.3. ERK1/2gene modification can help survival of BMSCs in cerebral ischemia rats, and transplantation of ERK1/2/BMSCs is better than only BMSCs in improving survival rate of neuron and astrocyte around margin of ischemia. |
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