CAJ | 학술논문

目的：研究Netrin-1对活体大鼠心肌缺血再灌注损伤（Ischemia reperfusion injury，I/R）的影响及机制。为临床上提供体外循环（Cardiopulmonary bypass，CPB）下心肌保护新的策略。方法雄性Spraghe-Dawley(SD)大鼠20只，体重450-500g，随机分成2组，体外循环预充液中添加Netrin-1（10只）[I/R+Netrin-1组]，体外循环预充液中不添加Netrin-1（10只）[I/R组]。建立无血预充大鼠深低温停循环（Deep hypothermic circulatory arrest，DHCA）的心肌缺血再灌注的模型。实验结束2h后处死大鼠留取心脏标本。在CPB前、CPB75min （复温后15min），CPB后2h留取大鼠动脉血测定心肌损伤指标[心肌肌钙蛋白I (cTn I)和心型脂肪酸结合蛋白(HFABP)]。实验结束取大鼠心脏标本用TUNEL法检测心肌细胞凋亡；用Western-blot法测定局部心肌组织直肠癌结肠癌缺失基因（Delete in colorectal carcinoma gene，DCC）蛋白指标。大体光镜下观察心脏组织病理形态学改变；大鼠CPB 前、CPB 15min、CPB 45min （停循环15min）以及 CPB 90min （复温30min）监测血气分析。结果 I/R+Netrin-1组较I/R组：血清中心肌肌钙蛋白I(cTn I)和心型脂肪酸结合蛋白(HFABP)含量明显减少；心肌缺血再灌注后心肌凋亡明显减少27.5%(P<0.05)；DCC表达增加(P<0.05)。HE染色镜下观察发现I/R+Netrin-1组心肌细胞损伤程度明显减轻。结论Netrin-1对心肌缺血再灌注损伤有保护作用，其机制可能与DCC有关。
목적：연구Netrin-1대활체대서심기결혈재관주손상（Ischemia reperfusion injury，I/R）적영향급궤제。위림상상제공체외순배（Cardiopulmonary bypass，CPB）하심기보호신적책략。방법웅성Spraghe-Dawley(SD)대서20지，체중450-500g，수궤분성2조，체외순배예충액중첨가Netrin-1（10지）[I/R+Netrin-1조]，체외순배예충액중불첨가Netrin-1（10지）[I/R조]。건립무혈예충대서심저온정순배（Deep hypothermic circulatory arrest，DHCA）적심기결혈재관주적모형。실험결속2h후처사대서류취심장표본。재CPB전、CPB75min （복온후15min），CPB후2h류취대서동맥혈측정심기손상지표[심기기개단백I (cTn I)화심형지방산결합단백(HFABP)]。실험결속취대서심장표본용TUNEL법검측심기세포조망；용Western-blot법측정국부심기조직직장암결장암결실기인（Delete in colorectal carcinoma gene，DCC）단백지표。대체광경하관찰심장조직병리형태학개변；대서CPB 전、CPB 15min、CPB 45min （정순배15min）이급 CPB 90min （복온30min）감측혈기분석。결과 I/R+Netrin-1조교I/R조：혈청중심기기개단백I(cTn I)화심형지방산결합단백(HFABP)함량명현감소；심기결혈재관주후심기조망명현감소27.5%(P<0.05)；DCC표체증가(P<0.05)。HE염색경하관찰발현I/R+Netrin-1조심기세포손상정도명현감경。결론Netrin-1대심기결혈재관주손상유보호작용，기궤제가능여DCC유관。
Objective To observe the effect and mechanism of Netrin-1on in vivo myocardial ischemia reperfusion injury in rats. For the clinic with myocardial protection about cardiopulmonary bypass with a new strategy. Metholds Twenty male Sprague-Dawley rats,weight (400-450)g,were randomly divided into two groups(each=10):I/R+Netrin-1 and I/R groups. To estab-lish of myocardial ischemia reperfusion model of no blood priming of rats with deep hypothermic circulatory arrest ( DHCA). Two groups of rats were given tracheal intubation in general anesthesia,mechanical ventilation,and cardiopulmonary bypass intubation. Blood samples were collected at the before of CPB,15min after rewarming and 2h after CPB for determination of myocardial injury markers of cardiac troponin I (cTn I) and heart fatty acid binding protein (HFABP). These was measured by Enzyme-linked im-munosorbent assay (ELISA). Heart samples were collected for detection of myocardial cell apoptosis with TUNEL method and de-tection of local myocardial DCC protein index with Western-blot method. hematoxylin and eosin (HE) staining of cardiac tissue specimens were observed in the Myocardium Pathological Changes. Blood samples were collected at the before of CPB ,15min after CPB and DHCA,at the termination of CPB and 2h after CPB for blood gas analysis. Results The I/R+Netrin-1group than in I/R group:serum cardiac troponin I (cTn I) and heart fatty acid binding protein (HFABP) expression was significantly reduced;myocar-dial apoptosis after myocardial ischemia-reperfusion injury decreased 23.9% (P<0.05);the expression of DCC increased (P<0.05). He staining microscopic observations showed that I/R+Netrin-1 group significantly reduced myocardial cell injury. At the same time,We found that I/R+Netrin-1 group of myocardial cell injury is reduced significantly through HE staining in light microscopy observation. Conclusion Netrin-1on myocardial ischemia reperfusion injury has a protective effect, its mechanism may be related with DCC.