| Objective: To observe the changes of brain damage in different time points after myocardial ischemia, and different time point acupuncture intervention on myocardial ischemia in rats hippocampus Bax, Bcl-2 protein changes, discusses the acupuncture intervention on myocardial ischemia in rat hippocampal neurons protective effects and mechanism, as to choose a better opportunity of acupuncture intervention and provide experimental basis in clinical, but also for the "heart controlling mind" theory to provide the experimental basis for studying.Methods:1. 60 healthy SD rats, weight about 220±20g Were randomly divided into sham operation group, immediately group, 24 h group, 3d group, 5d group with 8 rats in each group. The rest of conventional breeding, to supplement experiment. Rats in sham operation group only thoracotomy without coronary vascular ligation while rats in other groups were established to AMI model. The changes of organizational structure of the hippocampus were observed by light microscopy and the change of ultrastructure of hippocampus were observed by transmission electron microscopy(TEM) and the expression of Bax and Bcl-2 was measured by immunocytochemistry staining at different time points(immediately, 24 h,3d and 5d).2. 60 healthy SD rats, weight about 220±20g were Randomly divided into the sham operation group, model group, immediate acupuncture group, 24 h acupuncture group,3d acupuncture group,with 8 rats in each group. Rats in sham operation group don't ligate the left anterior descending branch of the coronary artery while rats in other groups were established to AMI model. Except the pseudo surgery group, the treatment group were electro acupuncture at Shenmen and Xinshu at different time points The sham operation group and the model group do not treated with acupuncture, while the other groups were treated with electro acupuncture after modeling, every time 30 min,once a day for 7 days. The rats were killed and the skeletal muscle was taken to observe the following indicators after 11 days: The changes of organizational structure of the hippocampus were observed by light microscopy; the expression of Bax and Bcl-2 was measured by immunocytochemistry staining; the expression of Bax mRNA and Bcl-2mRNA was measured by RT-PCR.Results:1.The extent of brain damage in rats caused by AMI at different time points.(1)Light microscope to observe changes of the hippocampus morphology in each group:pseudo operation group hippocampus neuron cell distribution is clear, in order and form more complete, nucleus was blue, oval, round or tapered and clear outline, the dyeing is uniform; model group appeared different degree of neuronal cell fuzzy distribution,malalignment, reduce cell, dyeing are not. Immediate group, 24 h group, 3d group, 5d group showed a gradually increasing trend.(2)The change of ultrastructure of hippocampus were observed by transmission electron microscopy(TEM) :The organizational structure of hippocampus cells in the sham operation group were complete, while the cells of hippocampus in the other groups were reduce, swelling and expansion and this change is continuous.(3)The expression of Bax and Bcl-2 was measured by immunocytochemistry staining:Positive expression mainly distributed in neural cells, mainly for small particles, some gathered themselves together. There is no difference between 3d group and 5d group(P>0.05), while There are have significant difference between other groups(P < 0.01).2.Effects of electron acupuncture at different time points on the change of Bax and Bcl-2 during the brain injury in rats caused by AMI.(1) Hippocampal morphological changes were observed under the light microscope:pseudo operation group of neurons distribution clearly and neatly arranged, form a complete, nucleus was blue shapes oval or circular cone shape, contour clear, dyeing uniformity; Model group of neurons distributed fuzzy, malalignment, reduce cell,dyeing uneven; the pathologic changes of acupuncture group were mitigated, which instantly acupuncture group improved the most obvious and 24 h acupuncture group, 3D acupuncture group improved gradually become bad.(2)The expression of Bax and Bcl-2 was measured by immunocytochemistry staining:Bax: pseudo operation group< immediately acupuncture group<24h acupuncture group<3d acupuncture group<5d acupuncture group; Bcl-2: pseudo operation group >immediately acupuncture group >24h acupuncture group >3d acupuncture group>5d acupuncture group. Bax and Bcl-2 respectively for each pairwise comparisons in addition to 3d acupuncture group and 5d acupuncture group Bax, bcl-2 have less significant external(P > 0.05). The rest of the group between pairwise comparisons were statistically significant(P < 0.01).(3)RT-PCR method to detect Bax mRNA and bcl-2 mRNA relative expression: test results and immunohistochemical method to detect consistent. Acupuncture can the and down regulated the expression of Bax mRNA levels in the expression of bcl-2 mRNA in hippocampus tissue was increased and can acupuncture group was significantly better than the 24 h acupuncture group, 3D needle acupuncture group, 24 h acupuncture group was better than the 3D acupuncture group.Conclusion:1.AMI can lead to brain injury, the degree of injury is increasing in immediately group,24h group, 3d group 5d group, there is no difference with 3d group and 5d group, the mechanism may be related to the expression of apoptosis factor Bax, Bcl-2.2.EA can protect neuron of hippocampus after brain injury in rats with AMI,the mechanism may be associated with reducing the expression of Bax and Bax mRNA and also have a relationship with increasing the expression of Bcl-2 and Bcl-2 mRNA.3.Acupuncture therapy should be implemented in the early of AMI, which is good for acupuncture protect neuron through reducing the expression of Bax and Bax mRNA and increasing the expression of Bcl-2 and Bcl-2 mRNA. |
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