TRPA1参与丙泊酚联合布比卡因所致背根神经节细胞损伤  

Transient receptor potential channel A1 is involved in propofol combined bupivacaine‑induced cellular injury to dorsal root ganglion neurons

作　　者：张鹏[1] 郭雯静 袁丹 喻旭娇 赖露颖[1] 徐世元[1] 李凤仙[1] Zhang Peng;Guo Wenjing;Yuan Dan;Yu Xujiao;Lai Luying;Xu Shiyuan;Li Fengxian(Department of Anesthesiology,Zhujiang Hospital,Southern Medical University,Guangzhou 510000,China;Department of Anesthesiology,the People's Hospital of Leshan,Leshan 614000,China)

机构地区：[1]南方医科大学珠江医院麻醉科,广州510000 [2]乐山市人民医院麻醉科,乐山614000

出　　处：《国际麻醉学与复苏杂志》2023年第11期1133-1138,共6页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金(81974192,82271392)。

摘　　要：目的:探讨非选择性阳离子通道瞬时受体电位通道A1(transient receptor potential channel A1, TRPA1)是否参与丙泊酚联合布比卡因所致背根神经节(dorsal root ganglion, DRG)细胞损伤。方法:以C57B6/J背景的野生型(wild type, WT)小鼠与TRPA1敲除(TRPA1^(-/-))小鼠为研究对象,分别采集小鼠腰段(L_(3)~L_(5))DRG进行原代细胞培养,培养24 h后进行体外细胞实验。取培养24 h的WT小鼠DRG细胞采用随机数字表法分为4组(每组9孔):对照组(未予以药物处理)、丙泊酚组(丙泊酚10 μmol/L)、布比卡因组(布比卡因2 mmol/L)、丙泊酚+布比卡因组(丙泊酚10 μmol/L、布比卡因2 mmol/L),观察WT小鼠DRG细胞在不同药物处理后的细胞形态变化。取培养24 h的WT小鼠DRG细胞采用随机数字表法分为3组(每组9孔):对照组(未予以药物处理)、丙泊酚+布比卡因组(丙泊酚10 μmol/L、布比卡因2 mmol/L)、HC-030031+丙泊酚+布比卡因组(丙泊酚10 μmol/L、布比卡因2 mmol/L、HC-030031 1 μmol/L),采用细胞计数试剂盒(cell counting kit-8, CCK-8)法检测DRG细胞活力并计算细胞存活率,MitoSOX Red超氧化物指示剂检测线粒体活性氧(reactive oxygen species, ROS)水平,JC-1法检测线粒体膜电位水平。WT小鼠与TRPA1^(-/-)小鼠DRG细胞根据小鼠基因型分为WT组与TRPA1^(-/-)组(每组9孔),比较两组DRG细胞在丙泊酚(10 μmol/L)联合布比卡因(2 mmol/L)处理后线粒体ROS水平及其膜电位的变化。 结果:与对照组、丙泊酚组、布比卡因组比较,布比卡因+丙泊酚组DRG细胞形态改变,甚至细胞破坏。与对照组比较,丙泊酚+布比卡因组细胞存活率降低( P<0.05),ROS水平升高( P<0.05),线粒体膜电位水平降低( P<0.05);与丙泊酚+布比卡因组比较,HC-030031+丙泊酚+布比卡因组细胞存活率升高( P<0.05),ROS水平降低( P<0.05),线粒体膜电位水平升高( P<0.05)。与WT组比较,TRPA1^(-/-)组在丙泊酚联合布比卡因处理后,DRG细胞线粒体ROS水平降低Objective To test whether the combined use of propofol and bupivacaine leads to cellular injury in dorsal root ganglion(DRG)neurons via a non‐selective cation permeable channel,the transient receptor potential channel A1(TRPA1).Meth‑ods Wild type(WT)mice and TRPA1 knockout(TRPA1^(−/−))mice with C57B6/J background were used as research subjects.DRG(L_(3)‒L_(5))from the lumbar segment of mice were collected for primary cell culture,and cultured for 24 h before conducting in vitro experiments.To observe the morphological changes of DRG cells in WT mice treated with different drugs,cells were randomly divided into 4 groups(n=9)using a random number table method:control group(no treatment),propofol group(10μmol/L),bupivacaine group(2 mmol/L),propofol+bupivacaine group(propofol 10μmol/L,bupivacaine 2 mmol/L).Using HC‐030031 to specifically block TRPA1,the effects of HC‐030031 on the cell activity and viability of WT mouse DRG cells induced by propofol combined with bupivacaine[cell counting kit 8(CCK‐8)],mitochondrial reactive oxygen species(ROS)levels(MitoSOX Red superoxide indicator detection),and membrane potential polarization(JC‐1 method)were measured.For this purpose,cells were divide into 3 groups using a random num‐ber table method(n=9):control group(no treatment),and propofol+bupivacaine group(propofol 10μmol/L,bupivacaine 2 mmol/L),HC‐030031+propofol+bupivacaine group(propofol 10μmol/L,bupivacaine 2 mmol/L,HC‐0300311μmol/L).Furthermore,we com‐pared the changes in mitochondrial ROS levels and membrane potential of DRG cells between WT mice(WT group)and TRPA1−/−mice(TRPA1^(−/−) group)(n=9)when using propofol(10μmol/L)combined with bupivacaine(2 mmol/L).Results Compared with the control group,propofol group,and bupivacaine group,the DRG cells in the bupivacaine+propofol group showed morphological changes and even cell destruction.Compared with the control group,the cell survival rate of the propofol+bupivacaine group decreased(P<0.05),the ROS level increased(P<0.05),and th

关 键 词：布比卡因 丙泊酚 瞬时受体电位通道A1 背根神经节 

分 类 号：R614[医药卫生—麻醉学]