针刺预处理对大鼠缺血心肌线粒体通透孔开放的调节机制的研究

发布时间：2018-03-14 06:17

本文选题：针刺预处理　切入点：缺血预处理　出处：《黑龙江中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过对大鼠缺血心肌线粒体通透性转换孔开放的调节研究,探讨针刺预处理抗心肌缺血的分子机制,并阐明针刺预处理对线粒体通透性转换孔的开放的联系;同时为本法用于临床防治缺血性心脏病和研究线粒体与电针预处理抗心肌缺血的关系提供实验科学依据。方法:将96只大鼠通过随机数字表相应分成空白组、模型组、针刺预处理组(针刺心俞穴)、缺血预处理组。采取活体大鼠心左冠状动脉前降支结扎方式复制心肌缺血及再灌注损伤模型。空白组:造模前,与针刺预处理组同固定大鼠30分钟但不针刺;每日1次、连续15日;不造模直接取材,整个实验过程不施加其他因素。模型组:造模前同空白组固定;心肌缺血及再灌注造模,整个实验过程不施加其他因素。针刺预处理组:造膜前15日开始针刺;每日一次,每次30min,共15日;心肌缺血及再灌注造模。缺血预处理组:造模前与空白组相同;造模时在进行冠状动脉结扎前先进行阻断血流5min,再灌流5min;如此连续反复3次,共30min的缺血预处理。心肌缺血造模成功后分别于即刻、24h、48h检测,血清CK、血清LDH、心肌缺血面积及心肌细胞线粒体膜通透性转换孔的开放程度。结果:1.模型组心肌梗死面积超过50%,血清CK、LDH检测值显著升高,激光共聚焦显微镜观察染色的线粒体绿色荧光与空白组对比明显减弱,实验结果表明其线粒体膜通透性转换孔活性显著减低(P0.01)。2.与模型组对照,缺血预处理组与针刺预处理即刻组的心肌梗死面积,血清CK、LDH均明显减低,激光共聚焦显微镜观察显示,线粒体荧光强度明显减低,但缺血预处理组与针刺预处理即刻组各项指标差异不显著(P0.05)。3.针刺预处理24h组、48h组与缺血预处理组及针刺预处理组即刻组对照心肌梗死面积、血清CK、LDH进一步减低;同时激光共聚焦显微镜观察到线粒体荧光强度有一定增强(P0.05或P0.01)。4.通过实验数据可以看出针刺预处理组与缺血预处理组均可治疗心肌缺血再灌注所引起的损伤;并且可能针刺预处理24h组为最佳治疗方案。结论:1.针刺预处理后即刻、24h、48h对缺血心肌的保护作用存在差异,并且治疗效果会随时间的不同而呈增强-峰值-减弱的趋势变化。2.针刺预处理与缺血预处理均可降低血清CK、LDH,减少再灌注损伤所引起的心肌梗死面积,从而减少心肌细胞凋亡。3.针刺预处理可能是通过抑制心肌缺血再灌注过程中mPTP的开放,从而减少线粒体的凋亡,减轻心肌缺血引起的损伤。4.针刺预处理可用于临床防治心肌缺血再灌注损伤,并且在临床治疗方面具有良好的疗效。
[Abstract]:Objective: to explore the molecular mechanism of acupuncture preconditioning in preventing myocardial ischemia by regulating the opening of mitochondrial permeability transition pore in ischemic myocardium of rats, and to elucidate the relationship between acupuncture preconditioning and opening of mitochondrial permeability transition pore. At the same time, it provides experimental scientific basis for clinical prevention and treatment of ischemic heart disease and the relationship between mitochondria and electroacupuncture preconditioning against myocardial ischemia. Methods: 96 rats were divided into blank group and model group by random digital table. Acupuncture preconditioning group (acupuncture at Xinshu acupoint, ischemic preconditioning group). The model of myocardial ischemia and reperfusion injury was established by ligating the anterior descending branch of left coronary artery in living rats. The rats in the acupuncture preconditioning group were fixed for 30 minutes without acupuncture; once a day, on 15th; no model was made directly and no other factors were applied throughout the experiment. Model group: fixed before modeling with blank group; model of myocardial ischemia and reperfusion, No other factors were applied in the whole experiment. Acupuncture preconditioning group: acupuncture began on 15th before membrane making; once a day for 30 minutes each time for 15th; myocardial ischemia and reperfusion modeling; ischemic preconditioning group: the model was the same as that in the blank group. Before coronary artery ligation, occlusion of blood flow was performed for 5 min and reperfusion for 5 min, then repeated for 3 times for 30 min. Myocardial ischemia was detected immediately at 24 h and 48 h after successful establishment of the model. Serum CK, serum LDH, myocardial ischemia area and opening degree of mitochondrial membrane permeability transition pore in myocardial cells. Results: the myocardial infarction area in the model group was more than 50, and the detection value of serum CK LDH was significantly higher than that in the model group. Compared with the blank group, the green fluorescence of mitochondria stained by laser confocal microscope was obviously weakened, and the results showed that the activity of mitochondrial membrane permeability transition pore was significantly decreased in the model group, compared with the model group. Myocardial infarction size and serum CK LDH in ischemic preconditioning group and acupuncture preconditioning group were significantly decreased, and the fluorescence intensity of mitochondria was significantly decreased by laser confocal microscopy. However, there was no significant difference between the ischemic preconditioning group and the acupuncture immediate preconditioning group (P 0.05). 3. The myocardial infarction size and the serum CKP LDH were further decreased in the 24h pretreatment group, the ischemic preconditioning group and the ischemic preconditioning group and the immediate acupuncture preconditioning group. At the same time, the fluorescence intensity of mitochondria was enhanced by laser confocal microscope. The experimental data showed that acupuncture preconditioning group and ischemic preconditioning group could treat myocardial ischemia-reperfusion injury. It is possible that 24 h acupuncture preconditioning group is the best therapeutic regimen. Conclusion 1. The protective effects of 24 h or 48 h acupuncture preconditioning on ischemic myocardium are different. And the therapeutic effect will change with time. 2.Acupuncture preconditioning and ischemic preconditioning can decrease the serum CKT LDH and reduce the myocardial infarction area caused by reperfusion injury. Acupuncture preconditioning may reduce the apoptosis of mitochondria by inhibiting the opening of mPTP during myocardial ischemia-reperfusion. Acupuncture preconditioning can be used to prevent and treat myocardial ischemia-reperfusion injury, and has a good effect in clinical treatment.
【学位授予单位】：黑龙江中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245

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