| Objective:In this experiment,the left middle cerebral artery occlusion(Middle Cerebral Artery Occlusion,MCAO)cerebral ischemia model was used to observe the effects of acupuncture,low-frequency rTMS and the combination of the two on the behavior of rats with cerebral ischemia,and on the cerebral cortex on the ischemic side The relative expression of BDNF and TrkB and the influence of changes in the number of neuronal dendritic spines,to explore the mechanism of neuroplasticity of acupuncture and rTMS in the treatment of ischemic stroke.Methods:60 SPF-grade healthy male SD rats were randomly divided into 5 groups: sham operation group,model group,acupuncture group,rTMS group and acupuncture combined rTMS group.The left side MCAO rat model was prepared by the suture method.After modeling,the inclusion and exclusion were performed according to Zea Longa’s method,and those with scores of 1-3 were included in this experiment.Intervention to MCAO rats started 24hours after the operation.The acupuncture group selected the following acupoints: "Baihui" and "Quchi","Neiguan","Hegu","Zusanli","Sanyinjiao","Shenmai" and "Zhaohai" points on the affected side;transcranial magnetic group: set stimulation The site is the M1 area of ??the contralateral side,1Hz,100% MT,stimulation time 3s,intermittent 2s.The operation group and model group only had the same degree of grasping intervention.7 days is a course of treatment,1 time/day,continuous intervention for 6 days,1 day of rest,2 weeks of intervention in total.MNSS(modified Neurological Severity Scores)score,Clark score rat general functional damage score,Clark score rat focal functional damage score were performed 24h,7,and 14d after modeling,and the cerebral cortex on the ischemic side was measured by Western Blot.The expression of BDNF and TrkB protein inside,Golgi-cox staining(Golgi-cox)measures the number of neuron dendritic spines.All experimental data used SPSS 23.0to create a database for statistical analysis.Results:1.m NSS score results:The m NSS score of the sham operation group before and after model building was 0.One day after the model was established,compared with the sham operation group,the m NSS score of the model group was significantly increased,with significant statistical difference(P<0.01);intervention 7,14days,the rats in each treatment group compared with the model group,the neurological function score All were significantly decreased,and there were statistical differences(P<0.05 or P<0.01).After 7 days of intervention,the score of the combined group was significantly lower than that of the acupuncture group(P<0.05);for 14days of intervention,the score of the combined group was significantly lower than that of the acupuncture group and rTMS group,with significant statistical difference(P<0.01).2.Clark score rat general functional impairment score result: Clark score rat general functional impairment score result: The Clark score rat general functional impairment score of the sham operation group before and after the model was all 0.1 day after modeling,compared with the sham operation group,the general functional injury score of the model group was significantly increased,with significant statistical difference(P<0.01);intervention 7,14d,acupuncture group,rTMS group and model group There was no statistical difference in comparison(P>0.05),but there was a downward trend.Compared with the model group,the rats in the acupuncture combined with rTMS group had a significant decrease in general functional impairment scores,with statistical differences(P<0.05).3.Clark score rat focal function damage score result: The Clark score of rats in the sham-operated group before and after model building were all 0.One day after the model was established,compared with the sham operation group,the focal functional injury score of the model group was significantly increased,with significant statistical difference(P<0.01);intervention 7,14d,each treatment group and the model group In comparison,the focal function damage scores were significantly reduced,and there were statistical differences(P<0.05 or P<0.01).After 14days of intervention,the scores of the combined group were significantly lower than those of the acupuncture group and rTMS group,with statistical differences(P<0.05 or P<0.01).4.Changes in the relative expression of BDNF and TrkB in the cerebral cortex of the ischemic side of rats in each group: Compared with the sham operation group,the expression of BDNF and TrkB in the cerebral cortex of the ischemic side of the model group decreased significantly,with significant statistical differences(P<0.01);Intervention for 7,14d,the relative expression of BDNF and TrkB in each treatment group were significantly up-regulated compared with the model group,with significant statistical differences(P<0.01).The expressions of BDNF and TrkB in the combined group were significantly up-regulated compared with the acupuncture group and rTMS group on the7 th and 14 th day of the intervention,with significant statistical difference(P<0.01).5.Changes in the number of neuronal dendritic spines in the cerebral cortex of the ischemic side of the rats in each group: Compared with the sham operation group,the number of neurons in the cerebral cortex of the ischemic side of the model group was significantly reduced,with significant statistical differences(P<0.01).After 7 and 14days of intervention,the number of neurons in the cerebral cortex of the ischemic side of the rats in each treatment group increased significantly compared with the model group,with significant statistical differences(P<0.01).Intervention for 7,14d,the number of neuronal dendritic spines in the combined group was significantly increased compared with the acupuncture group and rTMS group,with significant statistical difference(P<0.01).Conclusions:1.After the model was successfully established,the motor nerve function of the rats was significantly decreased.Acupuncture and rTMS intervention can all improve the symptoms of nerve defects in rats with cerebral ischemia to a certain extent;2.Under the intervention of acupuncture and rTMS,the mechanism of MCAO rats’ motor function recovery is related to the up-regulation of the expression of BDNF and TrkB in the cerebral cortex on the ischemic side,and the reconstruction and repair of brain structure and nerve function;3.Acupuncture and rTMS intervention can promote the regeneration of neuronal dendritic spines in the infarct focus and surrounding brain tissue,suggesting that acupuncture and rTMS intervention can promote the plasticity recovery of dendritic spines during the recovery period of cerebral infarction;4.The treatment effect of acupuncture combined with rTMS is the most significant,which is better than a single acupuncture and rTMS treatment. |
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