摘要
目的:观察麦粒灸预处理对大鼠心肌缺血再灌注损伤(MIRI)不同时期保护效应差异,探讨该影响与自噬相关蛋白Beclin 1表达的相关性。方法:SD大鼠随机分为模型组、麦粒灸1d组、麦粒灸2d组、麦粒灸3d组、麦粒灸4d组、麦粒灸5d组、麦粒灸7d组,每组6只。选择大鼠双侧"内关"穴进行麦粒灸预处理后,结扎心脏冠状动脉左前降支制备心肌缺血再灌注模型,进行心电图监测并于再灌注4h后取心脏,采用TTC染色评价不同时间预处理的效应差异。在明确麦粒灸预处理最佳干预时间后,增设假手术组、模型组、麦粒灸组、自噬抑制剂组,每组各12只,分别于缺血30min和再灌注240min各6只,ELISA法检测血清中肌钙蛋白(cTnT)含量,HE染色观察心肌病理学变化,Western blot法检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2同源的水溶性相关蛋白(Bax)、Beclin 1的表达水平。结果:与模型组比较,麦粒灸4、5、7d组大鼠心肌梗死面积百分比明显减少(P<0.05)。与假手术组相比,模型组心肌纤维断裂、排列紊乱,且有炎性细胞浸润,缺血期与再灌注期cTnT、Bax/Bcl-2、Beclin 1均升高(P<0.01)。与模型组相比,麦粒灸组与自噬抑制剂组心肌纤维断裂、排列紊乱程度较轻,间质水肿,少量炎性细胞浸润,缺血期与再灌注期cTnT、Bax/Bcl-2、Beclin 1含量均显著下降(P<0.01)。与自噬抑制剂组比较,麦粒灸组缺血期与再灌注期cTnT、Bax/Bcl-2、Beclin 1均升高(P<0.05)。各组再灌注期与缺血期相比,cTnT、Bax/Bcl-2、Beclin 1差异无统计学意义(P>0.05)。结论:麦粒灸"内关"穴能有效降低心肌梗死面积,该保护效应与自噬相关,其作用机制可能为促进缺血期自噬发生,抑制再灌注期自噬过表达,从而减轻MIRI。
Objective To observe the effect of moxibustion （Moxi） preconditioning with seed-sized moxa cones on myocardial ischemia-reperfusion injury （MI/RI） at different stages, and to analyze the correlation between this effect and the expression of autophagy related protein Beclin 1. Methods This study contains two parts： 1） changes of myocardial pathological injury and percentages of myocardial infarcted area at different time-points after modeling and Moxi intervention, and 2） effect of Moxi on contents of serum cardiac troponin T（cTnT） and expression of myocardial Bcl-2, Bax and Beclin 1 proteins. In the first part, 42 SD rats were randomly divided into model group, 1 day （d） Moxi group, 2 d Moxi group, 3 d Moxi group,4 d Moxi group, 5 d Moxi group and 7 d Moxi group. The model of MI/RI was established by ligating the left anterior descending coronary artery （LAD） for 30 min and reperfusion for 240 min. The electrocardiogram （ECG） of standard limb lead ][ was monitored and the heart was taken 4 h after reperfusion for examining myocardial infarcted size with triphenyl tetrazolium chloride （TTO） stai- ning. In the second part, 48 SD rats were randomized into sham-operation, model, moxibustion and autophagy inhibitor （3-MA） groups, with 12 rats in each group. The serum cTnT level was assayed and histopathological changes of the myocardial tissue below the ligation site were examined with HE staining, and the expression levels of Bcl-2, Bax and Beclin 1 proteins in the myocardial tissue below the LAD-ligated site were detected using Western blot. Results Compared with the model group, the percentages of myocardial infarcted area were significantly decreased in the 4 d, 5 d and 7 d Moxi groups （P〈0.05）, but without significant differences among the 3 Moxi groups （ P〉0.05）. The state of MI-induced breakage and disordered arrangement of myocardial fibers with interstitial edema and inflammatory cell infiltration at the MI stage in the Moxi group and at the reperfusion stage
出处
《针刺研究》
CAS
CSCD
北大核心
2017年第6期471-476,共6页
Acupuncture Research
基金
国家自然科学基金项目(No.81674063)
江苏省普通高校科研创新计划项目(No.KYLX 16-1152)
关键词
心肌缺血再灌注
麦粒灸
自噬
自噬相关蛋白Beclin
1
Myocardial ischemia reperfusion
Moxibustion with seed-size moxa cone
Autophagy
Autophagy associated protein Beclin 1
