线粒体途径细胞凋亡在脂肪乳剂减轻大鼠布比卡因心脏毒性中的作用  

Role of mitochondrion-dependent apoptosis in reduction of bupivacaine-induced cardiotoxicity by lipid emulsion in rats

作　　者：林婷婷[1] 朱晓娜 张竞雄[1] 夏芳芳[1] 陈鸿飞[1] 刘乐[1] 徐旭仲[1] Lin Tingting;Zhu Xiaona;Zhang Jingxiong;Xia Fangfang;Chen Hongfei;Liu Le;Xu Xuzhong(Department of Anesthesioloy,First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第一医院麻醉科,325000

出　　处：《中华麻醉学杂志》2019年第8期901-906,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(81670219);浙江省自然科学基金青年基金(LQ18H090006)。

摘　　要：目的评价线粒体途径细胞凋亡在脂肪乳剂减轻大鼠布比卡因心脏毒性中的作用。方法健康成年雄性SD大鼠45只,体重300~350 g,采用随机数字表法分成3组:假手术组(Sham组,n=5)、布比卡因组(B组,n=20)和脂肪乳剂组(L组,n=20)。经20 s静脉注射0.4%布比卡因30 mg/kg诱发心脏停搏,制备心脏毒性模型。复苏时,L组静脉注射20%脂肪乳剂5 ml/kg,B组静脉注射生理盐水负荷量5 ml/kg,随后均以1 ml·kg^-1·min-1速率静脉输注3 min。记录大鼠成功复苏和自主循环恢复(ROSC)120 min末生存情况。ROSC后每10 min记录SBP、HR、MAP、RPP、复跳后各时点RPP与基础值的比值(RPPh);记录开始给药到心脏停搏时间(To)、心肺复苏开始至出现第1个自主心跳的时间(Ts)和心肺复苏开始至ROSC出现的时间(Tr)。于ROSC 120 min时处死大鼠,取左心室组织,采用Western blot法检测Bax、Bcl-2、cleaved caspase-9、cleaved caspase-3及胞浆和线粒体细胞色素c(Cyt c)的表达;RT-PCR法检测Bax mRNA和Bcl-2 mRNA的表达;观察心肌超微结构。结果与Sham组比较,B组Bcl-2及其mRNA和线粒体Cyt c表达下调,Bax及其mRNA、cleaved caspase-9、cleaved caspase-3和胞浆Cyt c表达上调(P<0.05)。与B组比较,L组复苏成功率和生存率升高,Tr缩短,ROSC后SBP、HR、RPP和RPPh升高,Bcl-2及其mRNA和线粒体Cyt c表达上调,Bax及其mRNA、cleaved caspase-9、cleaved caspase-3和胞浆Cyt c表达下调(P<0.05),To与Ts差异无统计学意义(P>0.05),心肌病理学损伤减轻。结论脂肪乳剂减轻大鼠布比卡因心脏毒性的机制可能与抑制线粒体途径细胞凋亡有关。Objective To evaluate the role of mitochondrion-dependent apoptosis in reduction of bupivacaine-induced cardiotoxicity by lipid emulsion in rats.Methods Forty-five healthy adult male Sprague-Dawley rats,weighing 300-350 g,were divided into 3 groups by a random number table method:sham operation group(Sham group,n=5),bupivacaine group(B group,n=20),and lipid emulsion group(L group,n=20).Cardiac arrest was induced by intravenously injecting 0.4%bupivacaine 30 mg/kg over 20 s to establish the cardiotoxicity model.Twenty percent lipid emulsion was intravenously injected in a dose of 5 ml/kg during resuscitation in group L,and normal saline was intravenously injected in a loading dose of 5 ml/kg during resuscitation in group B,followed by a 3-min infusion of 1 ml·kg^-1·min^-1in two groups.The successful resuscitation and survival rate at 120 min of return of spontaneous circulation(ROSC)were recorded.Systolic blood pressure,heart rate,mean arterial pressure,rate-pressure product(RPP)and ratio of RPP at each time point after recovery of spontaneous heart beat to baseline value(RPPh)were recorded every 10 min after ROSC.The time from administration to cardiac arrest(T0),time from beginning of cardiopulmonary resuscitation to appearance of the first spontaneous heart beat(Ts)and time from beginning of cardiopulmonary resuscitation to appearance of ROSC(Tr)were recorded.Rats were sacrificed at 120 min of ROSC,and left ventricular tissues were obtained for determination of the expression of Bax,Bcl-2,cleaved caspase-9,cleaved caspase-3,cytochrome C(Cyt c)in cytoplasm and mitochondria(by Western blot)and expression of Bax and Bcl-2 mRNA(by real-time polymerase chain reaction)and for examination of myocardial ultrastructure.Results Compared with Sham group,the expression of Bcl-2 protein and mRNA and mitochondrial Cyt c was significantly down-regulated,and the expression of Bax protein and mRNA,cleaved caspase-9,cleaved caspase-3 and cytoplasmic Cyt c was up-regulated in B group(P<0.05).Compared with B group,the rate of su

关 键 词：脂肪乳剂 静脉注射用 布比卡因 药物相关性副作用和不良反应 线粒体 细胞凋亡 

分 类 号：R73[医药卫生—肿瘤]