摘要
目的：探讨针灸预处理对大鼠心肌缺血/再灌注（I/R）后的保护作用以及对高迁移率族蛋白B1（HMGB1）表达的影响。方法选择60只Wistar大鼠，按随机数字表法分为假手术组、心肌I/R模型组、针灸预处理组，每组20只。采用结扎冠状动脉左室支左心耳下缘约0.5 cm处阻断血流10 min后再灌注1 h制备I/R损伤模型；假手术组仅穿线不结扎；针灸预处理组于I/R前7 d给予每日1次电针内关穴20 min，连续治疗7 d。采用苏木素-伊红（HE）染色，光镜下观察心肌组织病理学变化，半定量积分法计算3组心肌组织病理学评分；采用酶联免疫吸附试验（ELISA）检测血浆HMGB1、肿瘤坏死因子-α（TNF-α）、心肌肌钙蛋白T（cTnT）的含量，逆转录-聚合酶链反应（RT-PCR）、蛋白质免疫印迹试验（Western Blot）检测心肌组织HMGB1、单核细胞趋化蛋白-1（MCP-1）、TNF-α的mRNA及蛋白表达水平。结果光镜下可见心肌I/R模型组心肌纤维部分断裂，心肌细胞大片状坏死，边界不清，细胞出现浓缩、破裂、溶解、甚至消失，间质水肿并伴大量炎性细胞浸润；针灸预处理组上述表现较心肌I/R模型组明显减轻。与假手术组比较，心肌I/R模型组HMGB1、TNF-α、cTnT含量和组织病理学评分均明显升高〔HMGB1（μg/L）：9.64±1.16比2.15±.031，TNF-α（μg/L）：91±22比19±5， cTnT（μg/L）：1.50±0.35比0.07±0.03，组织病理学评分（分）：2.5±0.3比0.0±0.0，均P＜0.01〕，HMGB1、MCP-1、TNF-αmRNA和蛋白表达均明显升高（HMGB1 mRNA：1.42±0.16比0.02±0.00，MCP-1 mRNA：0.46±0.06比0.01±0.00，TNF-αmRNA：0.75±0.04比0.03±0.00；HMGB1蛋白：1.08±0.01比0.02±0.01， MCP-1蛋白：0.92±0.03比0.40±0.01，TNF-α蛋白：1.10±0.02比0.35±0.01，P＜0.05或P＜0.01）；与心肌I/R模型组比较，针灸预处理组HMGB1（6.58±0.73）、TNF-α（63±19）、cTnT（1.15±0.31）含量均明显降低（均P＜0.01）
Objective To investigate the protective effect of electroacupuncture pretreatment on myocardial ischemia/reperfusion （I/R） injury and its influence on high mobility group box 1 （HMGB1） expression in rats. Methods Sixty Wistar rats were randomly divided into sham operation group, myocardial I/R model group and electroacupuncture pretreatment group by random number table （each n = 20）. Myocardial I/R injury model was reproduced by ligating the left ventricular branch coronary artery at about 0.5 cm below the atrial appendage lower margin for 10 minutes to occlude the blood flow, then the ligature was relaxed for 1 hour reperfusion; in electroacupuncture pretreatment group, 7 days before I/R, the electroacupuncture at Neiguan acupoint was applied once daily for 20 minutes till the 7th day when I/R was established. Under light microscope, the pathological changes of myocardial specimen stained by hematoxylin-eosine （HE） method were observed. The myocardial histopathological integral was detected by semi quantitative integral method, and the changes of histological scores in three groups were investigated. The levels of plasma HMGB1, tumor necrosis factor-α （TNF-α）, cardiac troponin T （cTnT） were detected by enzyme-labeled immunosorbent assay （ELISA）. The expressions of HMGB1, monocyte chemotactic protein-1 （MCP-1）, TNF-αmRNA and protein in myocardium were detected by reverse transcription-polymerase chain reaction （PT-PCR） and Western Blot. Results Under light microscope, the myocardial tissue in myocardial I/R model group showed partial fracture of myocardial fibers, large patches of myocardial cell necrosis, hazy boundary, cellular condensation, rupture and dissolution or even disappearance, interstitial edema with a lot of inflammatory cell infiltration; the above myocardial＆amp;nbsp;tissue injury in electroacupuncture pretreatment group was significantly milder than that in myocardial I/R model group. Compared with sham operation group, in myocardial I/R model group the HM
出处
《中国中西医结合急救杂志》
CAS
北大核心
2015年第1期33-37,共5页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
浙江省科技计划项目（2012C23087）
浙江省慢性病适宜技术示范基地项目（04-M-01）
浙江省嘉兴市科技计划项目（2012AY1070-1）
浙江省嘉兴市心血管重点学科基金项目（04-F-08）
浙江省嘉兴市重点科技创新团队项目（2014-JX-7）
