| Objective: To observe the effect of electroacupuncture on the expression of GAP-43 in the contralateral cerebral cortex and the collateral sprouting of the corticospinal nerve fibers in rats with unilateral focal cerebral infarction,and to investigate its relationship with the recovery of motor function in rats.Methods: A model of focal cerebral cortical infarcted rats was prepared by the method of Longa modified suture method by occlusion of middle cerebral artery occlusion(MCAO)by right common carotid artery occlusion.The neurological function of MCAO rats was evaluated according to the Bederson4-point method.The MCAO rats were selected as the experimental group.Three rats with MCAO were randomly selected and the brain tissue sections were stained with 2% TTC dosing at 24 h postoperatively to observe the position of cerebral infarction.Forty-eight MCAO rats were randomly divided into GAP-43 group and BDA group,and each group was divided into model group and electro-acupuncture group.A total of 48 rats in GAP-43 group were treated with4% paraformaldehyde perfusion at four time points after 3d,7d,14 d and 28 d after MCAO to observe the infarcts,and the expression and distribution of GAP-43 protein in the contralateral cerebral cortex were observed byimmunofluorescence staining.BDA group was divided into model group and electroacupuncture group,the two groups of 36 rats,and each group is divided into three subgroups.The intracranial injection of biotin dextran amine(BDA)was performed on the left cerebral cortex of rats at 24 h after 7 days and 14 days after MCAO.All subgroups were perfused with 4% paraformaldehyde at 14 days after injection(14 days,21 days,28 days after MCAO),and kill the rat to take the cervical cord tissue after perfusion.And then labeled BDA by immunohistochemical free bleaching staining for to observe the contralateral CST of the cervical spinal cord plane,and observe the number and distribution of fibers from the collateral sprouts of the CST nerve fibers to the infarcted cervical gray matter.Immunofluorescence double labeling method was used to observe the co-localization of BDA/GAP-43 in the infarcted cervical gray matter.The model 28 d group and electroacupuncture 28 d group of BDA group be evaluated the motor function of left forelimb in rats by footstep error test at 1day before operation and 3d,7d,14 d,21d,28 d after MCAO.Twelve healthy rats were randomly selected,and these rats were only isolated from common carotid artery,external carotid artery and internal carotid artery,no plug line,no ligation of blood vessels,respectively,as the sham-operation group of GAP-43 group and BDA group,each group 6 rats.Results:(1)After TTC staining and perfusion of 4% paraformal-dehyde,the infarct was seen on the right side of the brain,and the infarct range covered the cerebral cortex movement and sensory area.(2)Compared with the sham-operation group,the expression of GAP-43 in model group at the cerebral cortex ?,? layer increased significantly after MCAO 3d(P <0.05),continued to increase after 7d,the difference was statistically significant(P<0.05),and the positive expression of GAP-43 in cerebral cortex wassignificantly different between 7 days and 3d(P <0.05);compared with the model group,EA group cortex ?,? layer GAP-43-positive cells at each time point were all increased(P <0.05).(3)There was no significant difference in the motor function score between the electroacupuncture group and the model group before and after operation(P> 0.05),at other times significantly different(P <0.05);There was no significant difference in the total number of infarction contralateral CST fibers between the electroacupuncture group,the model group and the sham operation group(P> 0.05);Compared with the model group,the number of BDA-positive fibers in the gray-gray region of the infarcted side of the EA group increased significantly(P <0.05),Among them,especially in the14 d group(P <0.01),And BDA-positive fibers simultaneously express GAP-43;Bivariate correlation analysis showed that the improvement of motor function at each time point of electroacupuncture group was positively correlated with the number of CST collateral sprouting fibers(14d group,r = 0.867,P = 0.025;21d group,r = 0.841,P = 0.036;28d group,r = 0.864,P = 0.026).Conclusions:(1)The method of improving the middle cerebral artery occlusion of the middle cerebral artery can be used to prepare the model of focal cerebral cortex infarction rats,and has a certain degree of simplicity.(2)Electroacupuncture can promote the expression of GAP-43 protein in the contralateral cerebral cortex and the colony of CST in the cerebral infarction rats to a certain extent.(3)Rats with cerebral infarction infarcted contralateral CST collateral buds are involved in the recon-struction of motor function after cerebral infarction.Electroacupuncture has a positive effect on this reconstruction.(4)Promote the expression of GAP-43 to induce contralateralCST nerve fibers may be one of the mechanisms of electro-acupuncture in the treatment of motor dysfunction after cerebral infarction. |
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