经皮穴位电刺激对一次性大强度运动大鼠心肌细胞PKC的影响

发布时间：2018-05-03 14:30

本文选题：经皮穴位电刺激 + 一次性大强度运动　；参考：《武汉体育学院》2016年硕士论文

【摘要】：目的:观察经皮穴位电刺激干预后大鼠心肌细胞自噬、凋亡情况以及PKCδ、PKCε表达的变化,比较一次性大强度运动和不同形式的经皮穴位电刺激对大鼠心肌细胞的影响,探讨二者对运动心脏的作用机制,为中医传统方法在运动医学领域的应用提供理论依据。方法:健康雄性Sprague-Dauley(SD)大鼠24只,体重210±10 g,两月龄,随机分为4组,每组6只:无负重运动组(E)、6%负重运动组(EG)、6%负重加中频穴位电刺激预处理组(MEG)、6%负重加中频穴位电刺激后处理组(AEG),大鼠的运动时间为2小时。因一般运动员在休赛期都会保持小强度的训练,故本文将无负重小强度运动组(E)作为对照组。E组与EG组大鼠结束游泳运动后,立即断颈处死获取完整心脏;MEG组在游泳前半小时给予经皮穴位电刺激预处理,游泳结束后立即断颈处死,取完整心脏;AEG组在游泳后即刻给予穴位电刺激处理半小时,处理完后取材。刺激穴位为足三里和内关。通过HE染色、电镜观察心脏结构改变,运用免疫印迹试验(Western Blot)检测心肌细胞LC3II/I、Beclin1、m TOR、PKCδ、PKCε表达,Tunel检测心肌细胞凋亡。结果:1.一次性大强度运动后,大鼠心肌组织和细胞的结构形态产生一定变化,相比EG组经皮穴位电刺激的MEG组和AEG组的心肌肌纤维排列、线粒体形态、细胞核形态都有一定程度的改善,MEG组的细胞状态略好于AEG组,提示经皮穴位电刺激预处理对大鼠心肌结构保护的效果好于经皮穴位电刺激后处理。2.EG组的凋亡率显著高于E组(p0.01);MEG组的凋亡水平有显著的下调,AEG组的凋亡也出现下调(p0.01),但是下调的程度低于MEG组。Tunel检测结果提示,一次性大强度运动能够诱发心肌细胞的凋亡,而经皮穴位电刺激能够下调心肌组织的凋亡率,经皮穴位电刺激预处理的作用效果优于后处理。3.免疫印迹试验(Western Blot)结果显示,EG组的LC3II/I的比值显著高于E组(p0.05),提示大强度运动能够激活自噬,MEG组的表达高于EG组(p0.05),提示经皮穴位电刺激预处理能够进一步诱导自噬,对心脏起保护作用;AEG组的表达水平相较MEG组变化并不显著(p0.05)。心肌细胞Beclin1的表达情况,与E组相比,EG组的表达略有下降,但降幅较小,MEG组Beclin1的表达水平提高明显(p0.05),提示经皮穴位电刺激预处理可以上调Beclin1的表达,且具有显著差异(p0.05);AEG组表达量亦出现上升,并具有显著差异(p0.05),且AEG组和MEG组的表达水平差别不大,MEG组稍高一点,但差距并不明显(p0.05),提示经皮穴位电刺激预处理与后处理对心肌Beclin1表达影响作用差别可能并不大。心肌细胞m TOR的表达,与E组相比,EG组的表达显著下调(p0.01),而MEG组和AEG组的表达量相较于E组和EG组也均显著的下调(p0.01),AEG组的下降趋势比MEG组更显著(p0.01)。4.PKCδ的表达情况显示,相对E组,EG组PKCδ的表达上调;MEG和AEG组则呈下调趋势(p0.05),提示一次大强度运动能够上调PKCδ的表达,经皮穴位电刺激则能抑制其表达上调。心肌细胞PKCε的表达情况变化不明显,各组基本都处于同一水平,没有显著性变化(p0.05)。结论:1、一次性大强度运动(6%负重)可以同时激活大鼠心肌细胞的自噬与凋亡。2、经皮穴位电刺激可通过上调心肌细胞的自噬和抑制凋亡发生,对心肌进行保护。3、经皮穴位电刺激预处理下调心肌组织凋亡率的效果优于后处理,两者激活的自噬相关蛋白多无显著差异。4、一次性大强度运动能上调PKCδ的表达,经皮穴位电刺激预处理和后处理通过抑制大鼠心肌细胞PKCδ的过表达,发挥心肌保护作用。5、短周期一次性大强度运动和单次经皮穴位电刺激对PKCε表达无显著影响。
[Abstract]:Objective: To observe the autophagy, apoptosis and the changes of PKC Delta and PKC epsilon expression in rat cardiac myocytes after percutaneous acupoint electrical stimulation, and compare the effects of the single large intensity exercise and different forms of percutaneous acupoint electrical stimulation on the cardiac myocytes in rats, and explore the mechanism of the action of the two groups on the moving heart, and the traditional Chinese medicine method in the field of sports medicine. Methods: 24 healthy male Sprague-Dauley (SD) rats, weight 210 + 10 g, 2 month old, were randomly divided into 4 groups, and each group was randomly divided into 6 rats: no weight negative exercise group (E), 6% weight negative exercise group (EG), 6% load plus intermediate frequency acupoint electrical stimulation group (MEG), 6% weight negative plus intermediate frequency acupoint electrical stimulation group (AEG), and the exercise time of rats 2 hours. Because of the low intensity training of the general athletes during the period of the rest period, the small intensity exercise group (E) was used as the control group.E and the EG group to get the complete heart immediately after the end of the swimming exercise. In group MEG, the percutaneous point electrical stimulation was given half an hour before the swimming, and the neck was broken immediately after the swimming end. Death, the whole heart was taken, group AEG was given a half hour after the acupoint electric stimulation immediately after swimming. The stimulation points were Zusanli and Neiguan. The cardiac structural changes were observed by HE staining and electron microscopy. The expression of LC3II /I, Beclin1, m TOR, PKC Delta, PKC epsilon was detected by immunoblotting test (Western Blot), and the cardiomyocytes were detected by Tunel. Apoptosis. Results: 1. after a one-time high-intensity exercise, the structure and morphology of myocardial tissue and cells in the rat were changed to a certain extent. Compared with group EG, the myocardial fiber arrangement of the MEG group and AEG group, the morphology of mitochondria and the morphology of the nucleus were improved to a certain extent. The cell state of the group MEG was slightly better than that in the AEG group, suggesting the skin acupoint electricity. The effect of stimulation preconditioning on myocardial protection in rats was better than that of group.2.EG after percutaneous acupoint electrical stimulation. The apoptosis rate of group.2.EG was significantly higher than that in group E (P0.01). The apoptosis level of group MEG was significantly down, and the apoptosis of group AEG also decreased (P0.01), but the degree of downregulation was lower than that in MEG group.Tunel detection results. The apoptosis of cardiac myocytes was induced, and the percutaneous point electrical stimulation could reduce the apoptosis rate of myocardial tissue. The effect of percutaneous acupoint electrical stimulation was better than that of post processing.3. immunoblotting test (Western Blot). The ratio of LC3II/I in group EG was significantly higher than that in group E (P0.05), suggesting that high intensity exercise could activate autophagy and MEG group. It was higher than group EG (P0.05), suggesting that the preconditioning of percutaneous acupoint electrical stimulation could further induce autophagy and protect the heart. The expression level of AEG group was not significantly higher than that of the MEG group (P0.05). The expression of Beclin1 in cardiac myocytes was slightly lower than that of the E group, but the decrease was smaller, and the expression level of Beclin1 in MEG group was improved. P0.05, suggesting that the expression of Beclin1 can be up-regulated by preconditioning with acupoint electrical stimulation (P0.05), and the expression level of AEG group also rises, and has significant difference (P0.05), and the expression level of AEG and MEG groups is not very different, MEG group is slightly higher, but the gap is not obvious (P0.05), suggesting percutaneous acupoint electrical stimulation preconditioning. The expression of M TOR expression in cardiac myocytes was significantly lower than that in group E (P0.01), while the expression of MEG group and AEG group was significantly lower than that of the E group and EG group (P0.01), while the decline potential of the MEG group and AEG group was more significant than that in the group of MEG and AEG. The expression of the AEG group was more significant than that of the E group. The expression of the MEG TOR in the MEG group and AEG group was more significant than that in the group. The expression of PKC Delta in group EG was up-regulated in group E, and in group MEG and AEG was down downward trend (P0.05), suggesting that a large intensity exercise could increase the expression of PKC Delta, and the expression of MEG could be inhibited by skin acupoint electrical stimulation. The expression of PKC E in cardiac myocytes was not obvious, and there was no significant change in each group at the same level (P0.05). Conclusion: 1, 1 Secondary intensive exercise (6% weight negative) can activate autophagy and apoptosis of rat cardiac myocytes at the same time. The percutaneous acupoint electrical stimulation can protect myocardium by up regulation of autophagy and inhibition of apoptosis in cardiac myocytes. The effect of.2 on myocardial apoptosis rate is better than that of post-processing. The autophagy phase activated by both of them is better than that of postprocessing. There is no significant difference in.4, and the expression of PKC delta can be up-regulated by a one-time high-intensity exercise. The preconditioning and post-processing of percutaneous acupoint electrical stimulation can inhibit the overexpression of PKC Delta in rat cardiac myocytes and play the protective effect of.5. There is no significant effect on the expression of PKC e by short cycle one-time large intensity exercise and single transcutaneous electrical stimulation.

【学位授予单位】：武汉体育学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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