脂肪乳剂通过激活自噬流解救布比卡因心肌毒性  

Lipid emulsion rescues bupivacaine-induced myocardial toxicity by activating autophagic flux

作　　者：林婷婷[1] 施克俭[1] 金周晟[1] 张裕坚[1] 夏芳芳[1] 刘乐[1] LIN Tingting;SHI Kejian;JIN Zhousheng;ZHANG Yujian;XIA Fangfang;LIU Le(Department of Anesthesiology,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)

机构地区：[1]温州医科大学附属第一医院麻醉科,浙江温州325015

出　　处：《温州医科大学学报》2021年第8期609-614,共6页Journal of Wenzhou Medical University

基　　金：国家自然科学基金资助项目(81900231);温州市科技计划项目(Y20190418)-。

摘　　要：目的:探讨自噬在脂肪乳剂解救布比卡因心肌毒性中的作用。方法:取对数生长期的H9C2心肌细胞随机分成空白溶剂组(DMSO组)、布比卡因组(Bup组)、脂肪乳剂组(Lip组)、脂肪乳剂联用布比卡因组(BLp组),相应药物处理24 h后,CCK8检测细胞增殖抑制率,光镜下观察细胞形态,电镜下观察细胞自噬现象,激光共聚焦显微镜下监测自噬流情况,Western blot和RT-PCR检测LC3II/I、p62蛋白和mRNA表达水平。结果:与DMSO组相比较,Bup组心肌细胞增殖抑制率明显升高(P<0.05),光镜下可见细胞大量死亡皱缩,形态不规则,胞核不清,电镜下见自噬体大量堆积,激光共聚焦显微镜下见自噬流被抑制,LC3II/I、p62蛋白和mRNA表达水平均显著升高(P<0.05)。与Bup组相比较,BLp组与Lip组心肌细胞增殖抑制率显著下降(P<0.05);光镜下心肌死亡细胞减少,细胞界限清楚,胞浆清晰,胞核清楚;电镜下自噬体减少;自噬流加快自噬体清除;LC3II/I、p62蛋白和mRNA表达水平均显著下降(P<0.05)。结论:脂肪乳剂可通过激活自噬流来加快自噬体的清除从而解救布比卡因所致心肌毒性。Objective:To explore the role of autophagy in lipid emulsion to rescue myocardial toxicity of bupivacaine.Methods:The logarithmic growth of H9C2 cardiomyocytes were randomly divided into four groups:control group(DMSO group),bupivacaine group(Bup group),lipid emulsion group(Lip group),lipid emulsion+bupivacaine group(BLp group).After 24 hours of treatment with corresponding drugs,CCK8 was used to detect cell proliferation inhibition rate,cell morphology was observed under light microscope,cell autophagy was observed under electron microscope,and autophagy flow was detected under laser scanning confocal microscope.And we detected the expression of LC3II/I protein and p62 protein by Western blot and the expression of MAP1LC3 mRNA and p62 mRNA by RT-PCR in H9C2 cardiomyocytes.Results:Compared with the DMSO group,the Bup group significantly increased the myocardial cell proliferation inhibition rate(P<0.05),showed a large number of cell death and shrinkage,inhibited of autophagic flow,obviously increased of LC3II/I and p62 protein and mRNA expression(P<0.05).Compared with the Bup group,the BLp group and Lip group significantly reduced the myocardial cell proliferation inhibition rate(P<0.05),improved cardiomyocyte damage,promoted autophagic flow and accelerated the removal of autophagosomes,down regulated the expression of LC3II/I and p62 protein and mRNA(P<0.05).Conclusion:Lipid emulsion rescues bupivacaine-induced myocardial toxicity by activation autophagic flow,which accelerates the removal of autophagosomes.

关 键 词：脂肪乳剂 布比卡因 心肌毒性 自噬 

分 类 号：Q319.1[生物学—遗传学]