穴位埋线对缺血再灌注损伤高脂血症合并2型糖尿病大鼠心肌保护作用的机制研究

发布时间：2018-08-15 12:36

【摘要】：目的探讨穴位埋线对缺血再灌注损伤高脂血症合并2型糖尿病(ZDF)大鼠心肌保护作用的机制,为临床治疗提供理论依据。方法 2015年1月—2016年5月选取ZDF大鼠24只,随机分为对照组、缺血再灌注组、缺血预处理组和穴位埋线组,每组6只。适应性饲养1周后第7天,对照组直接开胸取材,其余3组均结扎冠状动脉左前降支,建立缺血再灌注损伤模型。缺血再灌注组缺血30 min,再灌注60 min;缺血预处理组缺血5 min、再灌注5 min,重复3次,之后操作同缺血再灌注组;穴位埋线组在适应性饲养1周后第1天,"内关""膻中""心俞"穴进行穴位埋线,之后操作同缺血再灌注组。采用透射电镜观察各组ZDF大鼠心肌组织超微结构及自噬水平,Western blotting法检测心肌内质网应激相关蛋白葡萄糖调节蛋白78(GRP78)、自噬相关蛋白Beclin-1水平,酶联免疫吸附实验(ELISA)法检测髓过氧化物酶(MPO)、丙二醛(MDA)水平。结果透射电镜下对照组线粒体和心肌细胞形态结构正常,未见明显自噬体;缺血再灌注组线粒体和心肌细胞肿胀明显,细胞膜破裂、内容物泄漏,仅见空泡或空腔及少量自噬泡;缺血预处理组、穴位埋线组与缺血再灌注组相比,心肌细胞超微结构损伤较轻,且自噬泡数量明显增多;缺血再灌注组心肌细胞超微结构破坏最为严重,且未见明显自噬泡。缺血再灌注组、缺血预处理组、穴位埋线组GRP78、Beclin-1、MPO、MDA水平高于对照组(P0.05);缺血预处理组、穴位埋线组GRP78、Beclin-1水平高于缺血再灌注组,但MPO、MDA水平低于缺血再灌注组(P0.05)。结论穴位埋线可能通过促进GRP78、Beclin-1水平升高,进而起到对再灌注损伤心肌的保护作用。
[Abstract]:Objective to investigate the mechanism of catgut embedding at acupoints on myocardial protection of hyperlipidemic rats with type 2 diabetes mellitus (ZDF) after ischemia-reperfusion injury, and to provide theoretical basis for clinical treatment. Methods from January 2015 to May 2016, 24 ZDF rats were randomly divided into three groups: control group, ischemia reperfusion group, ischemic preconditioning group and acupoint catgut embedding group with 6 rats in each group. On the 7th day after adaptive feeding, the patients in the control group were directly thoracotomy and the other three groups were ligating the left anterior descending branch of coronary artery to establish the model of ischemia reperfusion injury. The rats in the ischemia reperfusion group were subjected to ischemia for 30 minutes and reperfusion for 60 minutes, the ischemic preconditioning group for 5 minutes and the reperfusion group for 5 min, repeated for 3 times, and then operated the same as the ischemic reperfusion group. On the first day after adaptive feeding for 1 week, the acupoints of "Neiguan" Shanzhong "and" Xinshu "were treated with catgut embedding at the acupoint, and then operated the same as the ischemia-reperfusion group. The ultrastructure and autophagy level of myocardial tissue were observed by transmission electron microscope (TEM) in ZDF rats. The levels of glucose-regulated protein 78 (GRP78) and autophagy associated protein Beclin-1 (Beclin-1) were detected by Western blotting method. The level of malondialdehyde (MDA) of myeloperoxidase (MPO),) was detected by enzyme linked immunosorbent assay (ELISA). Results the morphology and structure of mitochondria and cardiomyocytes in the control group were normal under transmission electron microscope, and no autophagy was found in the control group, while in the ischemia reperfusion group, the mitochondria and myocardial cells were swollen, the cell membrane was ruptured, and the contents were leaking. Only cavitation or cavity and a small amount of autophagy were found in the ischemic preconditioning group and acupoint embedding group compared with the ischemia-reperfusion group the ultrastructural damage of myocardial cells was lighter and the number of autophagy was significantly increased. Myocardial ultrastructure damage was the most serious in ischemia reperfusion group, and no obvious autophagy was found. The MDA level of GRP78 Beclin-1 MPOO in ischemic preconditioning group, ischemic preconditioning group and acupoint embedding group was higher than that in control group (P0.05); in ischemic preconditioning group, GRP78 Beclin-1 level was higher than that in ischemia reperfusion group, but MPOO MDA level was lower than that in ischemia reperfusion group (P0.05). Conclusion Acupoint catgut implantation may protect myocardium from reperfusion injury by promoting the elevation of GRP78 and Beclin-1 levels.
【作者单位】：江苏大学附属医院针灸科;江苏大学职工医院;
【基金】：国家自然科学基金青年科学基金项目(81303037)
【分类号】：R245

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