摘要
目的观察电针预处理心俞穴和神门穴对大鼠离体心肌缺血再灌注心肌组织自噬的调控作用。方法将36只SD大鼠随机分为正常组、缺血再灌注组和电针预处理组,每组12只。电针预处理组在造模前3周给予电针心俞穴和神门穴干预,20 min/次,1次/d,6 d/周,连续干预3周后利用离体灌流装置制备大鼠离体心肌缺血再灌注模型(平衡灌注20 min,缺血40 min,再灌注2 h);缺血再灌注组不给予术前干预,模型制备方法同电针预处理组;正常组无缺血再灌注操作,平衡灌注180 min。停灌注前20 min取冠状动脉流出液,检测心肌酸激酶(CK)、心肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)含量;灌流结束后,苏木精-伊红(HE)染色观察大鼠心肌组织病理形态,透射电镜观察大鼠心肌组织超微结构,Western blot法检测大鼠心肌组织中自噬标志蛋白Beclin-1和LC3-Ⅱ表达情况。结果缺血再灌注组冠状动脉流出液中CK、CK-MB、LDH含量均显著高于正常组(P均<0.05),电针预处理组均显著低于缺血再灌注组(P均<0.05)。HE染色显示缺血再灌注组心肌纤维溶解、断裂,甚至坏死,细胞肿胀;电针预处理组心肌纤维间隙轻度增宽,坏死灶少,细胞肿胀程度较轻。透射电镜显示缺血再灌注组心肌细胞线粒体嵴排列不规则、断裂,自噬溶酶体数量显著增加;电针预处理组心肌细胞核及线粒体结构基本完整,自噬溶酶体数量较缺血再灌注组少。缺血再灌注组心肌组织中Beclin-1和LC3-Ⅱ表达量均显著高于正常组(P均<0.05),电针预处理组均显著低于缺血再灌注组(P均<0.05)。结论电针预处理可通过下调自噬标志蛋白的表达,从而发挥保护心肌、减轻大鼠心肌缺血再灌注损伤的作用。
Objective It is to observe the regulatory effect of electroacupuncture at Xinshu acupoint(Bl-15)and Shenmen acupoint(HT-7)on autophagy of isolated myocardial ischemia-reperfusion in rats.Methods Thirty-six SD rats were randomly divided into normal group,ischemia-reperfusion group and electroacupuncture preconditioning group,with 12 rats in each group.The electroacupuncture preconditioning group was treated with electro-acupuncture at Xinshu and Shenmen acupoints 3 weeks before modeling,20 min/time,once/d,6 d/week.After continuous intervention for 3 weeks,the isolated rat myocardial ischemia-reperfusion models(balanced perfusion for 20 min,ischemia for 40 min,and reperfusion for 2 h)were established by Langendorff device in the electroacupuncture preconditioning group;no preoperative intervention was given in the ischemia-reperfusion group,and the model preparation method was the same as the electroacupuncture preconditioning group;no ischemia-reperfusion operation was performed in the normal group,balanced perfusion for 180min.Coronary perfusate was collected 20 min before stopping perfusion,the contents of cardiac creatine kinase(creatinekinase,CK),myocardial creatinekinase isoenzyme(CK-MB)and lactatedehydrogenase(LDH)in the perfusate were measured;after perfusion,the pathological morphology of rat myocardial tissue was observed by hematoxylin-eosin(HE)staining,and the ultrastructure of rat myocardial tissue was observed by transmission electron microscope,the expression of autophagy marker protein Beclin-1,LC3-Ⅱin myocardial tissue of rats in each group was detected by Western blotting.Results The contents of CK,CK-MB and LDH in coronary persusate in the ischemia-reperfusion group were significantly higher than those in the normal group(all P<0.05),and these indexes in the electroacupuncture preconditioning group were significantly lower than those in the ischemia-reperfusion group(all P<0.05).HE staining showed that fibrinolysis and rupture,even necrosis of myocardial tissue,and cell swelling were found in the ischemia-reperfusion group;the myocardial fiber space was slightly widened,with fewer necrotic foci and less cell swelling in the electroacupuncture pretreatment group.Transmission electron microscopy showed that the mitochondrial cristae of cardiomyocytes were irregularly arranged and fractured,and the number of autophagy-lysosomes increased significantly in the ischemia-reperfusion group;the structures of the nucleus and mitochondria of cardiomyocytes in the electroacupuncture-reperfusion group were basically intact,and the number of autophagy-lysosomes was less than that in the ischemia-reperfusion group.The expression levels of Beclin-1 and LC3-Ⅱin myocardial tissue in the ischemia-reperfusion group were significantly higher than those in the normal group(all P<0.05),and these indexes in the electroacupuncture preconditioning group was significantly lower than that in the ischemia-reperfusion group(all P<0.05).Conclusion Electroacupuncture preconditioning can protect the myocardium and reduce myocardial ischemia-reperfusion injury in rats by down-regulating the expression of autophagy marker proteins.
作者
尹侠
李宏玉
朱路文
唐强
YIN Xia;LI Hongyu;ZHU Luwen;TANG Qiang(Heilongjiang University of Traditional Chinese Medicine, Harbin 150040, Heilongjiang, China;The Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine, Harbin 150001, Heilongjiang, China)
出处
《现代中西医结合杂志》
CAS
2022年第12期1607-1611,1616,共6页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
黑龙江省中医药管理局科研基金项目(ZHY18-068)
黑龙江省博士后基金项目(LBH-Z20201)
黑龙江中医药大学研究生创新基金(2022yjscx007)。
