套线技术制备小鼠心肌缺血/再灌注损伤模型

Preparation of myocardial ischemia/reperfusion injury model by overlapping line in mice

目的提高小鼠心肌缺血再灌注损伤(I/R)模型的手术速度和可靠性。方法比较3种模型技术的手术时间和小鼠存活率,检测术后白细胞计数和血清IL-6水平评估炎性反应,用TTC/Evans blue染色评估梗死面积,用压力导管检测左心室舒张末压和压力变化速率评估心功能。传统技术在气管插管呼吸机辅助通气支持下开胸结扎、松开左冠状动脉(LCA);新型技术用异氟烷吸入麻醉,经胸壁挤出心脏活结结扎LCA,抽开活结松开LCA;套线技术同样经胸壁挤出心脏,但用方结同时结扎LCA及1根2-0尼龙线,之后抽出该尼龙线松开LCA。结果与传统技术比较,新型技术和套线技术总手术时间明显缩短(P<0.01),血白细胞计数和血清IL-6水平降低(P<0.01或P<0.05),小鼠存活率提高(P<0.05);与新型技术比较,套线技术简化手术操作并缩短总手术时间(P<0.01)。3组小鼠心脏梗死面积和心功能差异无统计学意义(P>0.05),但套线组和传统组的变异系数低于快速组。结论套线技术降低手术难度,较新型技术进一步缩短手术时间并提高了小鼠I/R模型的稳定性。

Objective To improve the operation speed and reliability of the myocardial ischemia/reperfusion injury（I/R）model in mice.Methods The operative time and the survival rate of the three models were compared.The postoperative leukocyte count and serum IL-6 level were detected to evaluate the inflammatory response.The infarct size was assessed by TTC/Evans blue staining.The left ventricular end diastolic pressure and the pressure change rate were measured by apressure catheter for the evaluation of cardiac function.In traditional method,open chest surgery was performed to ligate and loose the left coronary artery（LCA）under the assistance of intubation and ventilation.In novel method,isoflurane was used for anesthesis,and then heart was squeezed out of the chest to ligate and loose LCA with a slipknot.In overlapping line method,the heart was also squeezed out,but a square knot was performed to ligate LCA and a 2-0 nylon together,and then the nylon thread was drew out to loose LCA.Results Compared with traditional method,the novel method and overlapping line method had significant short operation time（P〈0.01 or P〈0.05）,less white blood cell count and serum IL-6 concentration（P〈0.05）and higher mice survival rate（P〈0.01）;compared with novel method,the overlapping line method further simplified the reperfusion procedure and decreased total procedure time（P〈0.01）.There was no significant difference in cardiac infarct size and cardiac function by using the three methods（P〈0.05）,but the overlapping line method had less coefficient of variation compared to novel method.Conclusion Compared with the novel method,the overlapping line method reduces the operation difficulty,shortens the operation time and improves the stability of I/R model.