文献类型：期刊文献

中文题名：脂肪乳对布比卡因中毒大鼠心肌线粒体能量代谢损伤的影响

英文题名：Effect of lipid emulsion on myocardial mitochondrial energy metabolism injury in bupivacaine intoxicated rats

作者：王春爱[1];李玉兰[3];于妍[2];秦晓宇[4];温晓辉[5]

第一作者：王春爱

机构：[1]甘肃省中医院麻醉科,兰州市730050;[2]甘肃省中医院心血管科,兰州市730050;[3]兰州大学第一医院麻醉科;[4]甘肃中医药大学第一临床学院;[5]甘肃中医药大学免疫教研室

第一机构：甘肃省中医院麻醉科,兰州市730050

年份：2021

卷号：37

期号：9

起止页码：964

中文期刊名：临床麻醉学杂志

外文期刊名：Journal of Clinical Anesthesiology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2021_2022】；

基金：国家自然科学基金(81760892)。

语种：中文

中文关键词：脂肪乳;布比卡因;线粒体;心肌;能量代谢

外文关键词：Lipid emulsion;Bupivacaine;Mitochondria;Myocardial;Energy metabolism

摘要：目的探讨脂肪乳对布比卡因中毒大鼠心肌线粒体能量代谢损伤的影响。方法雄性Wistar大鼠40只,6月龄,体重250~300 g。采用随机数字表法分为四组:空白对照组(C组)、布比卡因中毒模型组(B组)、脂肪乳治疗组(T组)、脂肪乳预防组(P组),每组10只。C组经股静脉输注生理盐水3 ml/kg;B组经股静脉输注0.5%布比卡因10 mg/kg,出现心律失常后立即输注生理盐水3 ml/kg;T组经股静脉输注0.5%布比卡因10 mg/kg,出现心律失常后立即输注20%脂肪乳3 ml/kg;P组经股静脉输注20%脂肪乳3 ml/kg后5 min,输注0.5%布比卡因10 mg/kg。C组在开始输注生理盐水的第8分钟,B组、T组和P组在开始输注布比卡因的第8分钟处死大鼠。取左室心肌组织,分离心肌细胞线粒体。采用ELISA法测定心肌线粒体ATP酶(ATPase)活性,磷钼酸比色法测定ATP浓度,荧光比色法测定线粒体膜电位(MMP),Western blot法测定心肌线粒体过氧化物酶体增生物激活受体γ辅激活因子-1α(PGC-1α)、核呼吸因子1(NRF1)、线粒体转录因子A(mtTFA)蛋白含量,透射电镜观察线粒体结构。结果与C组比较,B组、T组和P组心肌线粒体ATPase活性、ATP浓度、MMP和PGC-1α、NRF1蛋白含量均明显降低(P<0.05),B组和T组心肌线粒体mtTFA蛋白含量均明显降低(P<0.05),P组心肌线粒体mtTFA蛋白含量差异无统计学意义。与B组比较,T组和P组心肌线粒体ATP浓度、MMP、PGC-1α、NRF1、mtTFA蛋白含量均明显升高(P<0.05),P组心肌线粒体ATPase活性均明显升高(P<0.05),T组心肌线粒体ATPase活性差异无统计学意义。与T组比较,P组心肌线粒体ATPase活性、PGC-1α、NRF1、mtTFA蛋白含量均明显升高(P<0.05),心肌线粒体ATP浓度和MMP差异无统计学意义。电镜下C组心肌纤维排列紧密,线粒体丰富,包膜完整;B组心肌纤维排列紊乱,线粒体分布不均匀,部分线粒体变形;T组和P组心肌纤维排列尚可,线粒体较丰富,部分线粒体变形。结论脂肪乳可以升高心肌线粒体ATPase活性、ATP浓度、MMP及线粒体内ATP合成相关蛋白PGC-1α、NRF1、mtTFA含量,减轻布比卡因中毒大鼠心肌损伤,预防使用脂肪乳效果更优。
Objective To investigate the effect of lipid emulsion on the damage of myocardial mitochondrial energy metabolism in bupivacaine intoxicated rats.Methods Forty male Wistar rats were 6 months old,weighing 250-300 g.By random number table method,they were divided into four groups:blank control group(group C),bupivacaine poisoning model group(group B),lipid emulsion treatment group(group T)and lipid emulsion prevention group(group P),10 rats in each group.Group C was infused with normal saline 3 ml/kg via femoral vein.Group B was infused with 0.5%bupivacaine 10 mg/kg via femoral vein and 3 ml/kg normal saline immediately after arrhythmia.Group T was infused with 0.5%bupivacaine 10 mg/kg via femoral vein,and 20%lipid emulsion 3 ml/kg immediately after arrhythmia occurred.Group P was infused with 20%lipid emulsion 3 ml/kg via femoral vein and 0.5%bupivacaine 10 mg/kg 5 minutes later.The rats in group C were sacrificed aT_(8)minutes after the infusion of normal saline,while the rats in groups B,T and P were sacrificed aT_(8)minutes after the infusion of bupivacaine.Left ventricular myocardial tissue was taken and mitochondria of myocardial cells were isolated.Mitochondrial ATPase activity was determined by ELISA,ATP concentration was determined by phosphomolybdic acid colorimetry,mitochondrial membrane potential(MMP)was determined by fluorescence colorimetry,the protein contents of peroxisome-proliferator-activated receptorγcoactivator-1α(PGC-1α),nuclear respiratory factor 1(NRF1)and mitochondrial transcription factor A(mtTFA)were determined by Western blot,and the mitochondrial structure was observed by transmission electron microscopy.Results Compared with group C,the myocardial mitochondrial ATPase activity,ATP concentration,MMP,PGC-1αand NRF1 protein contents in groups B,T and P were significantly decreased(P<0.05),and the myocardial mitochondrial mtTFA protein content in groups B and T was significantly decreased(P<0.05).There was no significant difference in the content of myocardial mitochondrial mtTFA protein in group P.Compared with group B,myocardial mitochondrial ATP concentration and protein contents of MMP,PGC-1α,NRF1 and mtTFA in groups T and P were significantly increased(P<0.05),and myocardial mitochondrial ATPase activity in group P was significantly increased(P<0.05),but there was no statistical significance in the activity of myocardial mitochondrial ATPase in group T.Compared with group T,myocardial mitochondrial ATPase activity,PGC-1α,NRF1 and mtTFA protein contents in group P were increased(P<0.05),but there was no statistical significance in myocardial mitochondrial ATP concentration and MMP.Under electron microscope,myocardium fibers in group C were closely arranged,with abundant mitochondria and complete capsule.In group B,myocardial fibers were disordered,mitochondria were not evenly distributed,and some mitochondria were deformed.The myocardial fibers in groups T and P were arranged fairly well,with abundant mitochondria,and some mitochondria were deformed.Conclusion lipid emulsion can increase myocardial mitochondrial ATPase activity,ATP concentration,MMP and the contents of mitochondrial ATP synthesis related proteins PGC-1α,NRF1 and mtTFA,and reduce the myocardial injury of bupivacaine intoxicated rats,while using lipid emulsion preventively is better.