大鼠离体心脏心肌胰岛素抵抗实验模型的建立  

Establishment of myocardial insulin resistance model in rat isolated hearts

作　　者：惠永鹏 巫洪坤 潘斯斯 陈开远 梁贵友 Hui Yongpeng;Wu Hongkun;Pan Sisi;Chen Kaiyuan;Liang Guiyou(Department of Cardiovascular Surgery,Affiliated Hospital of Guizhou Medical University,Guiyang 550001,China)

机构地区：[1]贵州医科大学附属医院心血管外科,贵阳550001

出　　处：《中华实验外科杂志》2023年第8期1641-1644,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81960051、82170286)。

摘　　要：目的建立大鼠离体心脏缺血再灌注实验模型并明确其诱导心肌胰岛素抵抗的效果。方法 12只雄性Sprague-Dawley(SD)大鼠由贵州医科大学实验动物中心提供, 按随机数字表法分为两组, 正常对照(NC)组(6例)离体心脏持续灌注60 min, 缺血再灌注(I/R)组(6例)全心缺血30 min, 再灌注30 min, 记录心功能指标, 检测肌酸激酶MB同工酶(CK-MB)和心肌肌钙蛋白T(cTnT)含量, 苏木精-伊红染色观察心肌组织细胞形态, 蛋白质免疫印迹和组织免疫荧光检测葡萄糖转运蛋白4(GLUT4)的表达和转位情况。组间比较用独立样本t检验。结果 I/R组心功能指标[左心室发展压(LVDP)(50.8±5.5) mmHg(1 mmHg=0.133 kPa)、左心室内压最大上升速率(+dp/dtmax)(2 566.6±358.6) mmHg/s、左心室内压最大下降速率(-dp/dtmax)(1 172.6±121.2) mmHg/s]明显低于NC组[LVDP(79.9±4.0) mmHg、+dp/dtmax(3 587.8±334.5) mmHg/s、-dp/dtmax(2 243.8±252.4) mmHg/s, t=9.625、4.656、8.556, P<0.05]。I/R组心肌酶[CK-MB(1.018±0.132) ng/ml、cTnT(47.791±3.815) pg/ml]明显高于NC组[CK-MB(0.630±0.080) ng/ml、cTnT(29.036±5.319) pg/ml, t=5.005、5.731, P<0.05]。与NC组比较, I/R组的心肌组织细胞肿胀明显、间隙增大、排列紊乱、部分心肌纤维中断。I/R组GLUT4膜蛋白(M-GLUT4)的相对表达量(0.234±0.016)显著低于NC组(0.482±0.034, t=11.420, P<0.05)。结论利用Langendorff灌注系统可以成功建立大鼠离体心脏缺血再灌注心肌胰岛素抵抗实验模型。Objective To establish an experimental model of ischemia/reperfusion(I/R)in rat isolated hearts and define the effects in inducing myocardial insulin resistance.Methods Male Sprague-Dawley(SD)rats were provided by the Experimental Animal Center of Guizhou Medical University,and divided into two groups by the random number table method.In the normal control(NC)group(6 cases),the isolated hearts were continuously perfused for 60 min.In the I/R group(6 cases),the whole hearts ischemia was performed for 30 min and reperfusion for 30 min.Cardiac function indicators were recorded.The contents of creatine kinase MB isoenzyme(CK-MB)and cardiac troponin T(cTnT)were detected.Cell morphology of myocardial tissues was observed by hematoxylin-eosin staining.The expression and translocation of glucose transporter 4(GLUT4)were detected by Western blotting and tissue immunofluorescence.Independent sample t-test was used for comparison between groups.Results The indicators of cardiac function in the I/R group[left ventricular developed pressure(LVDP):(50.8±5.5)mmHg(1 mmHg=0.133 kPa);maximum increase rate of left ventricular pressure(+dp/dtmax):(2566.6±358.6)mmHg/s;maximum decrease rate of left ventricular pressure(-dp/dtmax):(1172.6±121.2)mmHg/s]was significantly lower than those in the NC group[LVDP:(79.9±4.0)mmHg;+dp/dtmax:(3587.8±334.5)mmHg/s;-dp/dtmax(2243.8±252.4)mmHg/s,t=9.625,4.656,8.556,P<0.05].Myocardial enzymes[CK-MB(1.018±0.132)ng/ml,cTnT(47.791±3.815)pg/ml]in the I/R group were significantly higher than those in the NC group[CK-MB(0.630±0.080)ng/ml,cTnT(29.036±5.319)pg/ml,t=5.005,5.731,P<0.05].Compared with the NC group,cardiomyocytes swelling,increased intercellular,disorganized permutation and interrupted partial myocardial fibers of myocardial tissue were obviously observed in the I/R group.The relative expression of membrane protein of GLUT4(M-GLUT4)in the I/R group(0.234±0.016)was significantly lower than that in the NC group(0.482±0.034,t=11.420,P<0.05).Conclusion The Langendorff perfusion system can

关 键 词：离体心脏 心肌胰岛素抵抗 心肌缺血再灌注损伤 

分 类 号：R-332[医药卫生] R54