布比卡因致心肌细胞线粒体损伤大鼠模型的建立

Establishment of a rat model of myocardial mitochondrial damage induced by bupivacaine

目的观察电刺激下布比卡因心肌细胞线粒体形态变化及活性氧(reactive oxygen species, ROS)生成量,探讨建立理想的布比卡因中毒的大鼠心肌细胞模型。方法采用Langendroff装置新鲜分离雄性SD大鼠心肌细胞,细胞计数后将其转移至doff管中随机分为四组:DMEM静置组、DMEM电刺激组、布比卡因静置组、布比卡因电刺激组。实验重复五次。采用透射电镜观察心肌细胞线粒体形态,并使用多功能微孔板检测仪测量ROS生成量。结果 DMEM电刺激组线粒体肿胀程度及ROS生成量与DMEM静置组相比差异无显著性(P> 0.05);而布比卡因电刺激组线粒体肿胀程度明显高于布比卡因静置组(P=0.000),且ROS生成量也明显升高(P<0.05)。结论电刺激下心肌细胞呈节律性收缩,能更好地模拟临床布比卡因中毒时心肌线粒体损伤。

Objective To observe the morphological changes of cardiomyocytic mitochondria and the reactive oxygen species content in bupivacaine-exposed myocardial cells under electrical stimulation, and to establish an ideal myocardial cell model of bupivacaine poisoning. MethodsA Langendorff device was used to separate the cardiomyocytes of male SD rats. The cells were divided into four groups randomly: a DMEM static group, DMEM electric stimulation group, bupivacaine static group and bupivacaine plus electric stimulation group. The experiment was repeated for five times. The cardiomyocytic mitochondrial morphology was observed by transmission electron microscopy, and the ROS content was measured with a multifunctional microplate detector. ResultsThe degree of mitochondrial swelling and the ROS content in the DMEM electric stimulation group were not significantly different from those of the DMEM group ( P > 0.05), but the mitochondrial swelling in the bupivacaine plus electric stimulation group was significantly higher than that of the bupivacaine group ( P =0.000), and the ROS output was also significantly increased ( P < 0.05). ConclusionsUnder electrical stimulation, cardiomyocytes show rhythmic contractions, allowing better simulation of the myocardial mitochondrial injury during clinical bupivacaine poisoning.