盐酸右美托咪定对2-氯乙基乙基硫醚致人支气管上皮细胞损伤的保护作用及机制  

Mechanism of dexmedetomidine hydrochloride on 2-chloroethyl ethyl sulfide-induced injury in human bronchial epithelial cells

作　　者：师敏婕 李嘉 邓紫丰 詹天豪 刘建豪 赵昱舜 李文丽 刘江正 孔德钦 SHI Minjie;LI Jia;DENG Zifeng;ZHAN Tianhao;LIU Jianhao;ZHAO Yushun;LI Wenli;LIU Jiangzheng;KONG Deqin(Department of Military Toxicology and Chemical Defense Medicine,School of Military Preventive Medicine,Air Force Medical University,Key Laboratory of Free Radical Biology and Medicine of Shaanxi Province,Key Laboratory of Environmental Hazard Assessment and Prevention of Special Operations of Ministry of Education,Xi'an 710032;School of Public Health,Shaanxi University of Chinese Medicine,Xi'an 712046;Department of Recuperation and Rehabilitation for Flight Personnel,School of Aerospace Medicine,Air Force Medical University,Xi’an 710032,Shaanxi,China)

机构地区：[1]空军军医大学军事预防医学系军事毒理学与防化医学教研室,特殊作业环境危害评估与防治教育部重点实验室,陕西省自由基生物学与医学重点实验室,陕西西安710032 [2]陕西中医药大学公共卫生学院,陕西西安712046 [3]空军军医大学航空航天医学系飞行人员康复与疗养教研室,陕西西安710032

出　　处：《癌变．畸变．突变》2024年第5期342-348,358,共8页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(32100996);陕西省重点研发计划(2023-YBSF-297)。

摘　　要：目的:研究盐酸右美托咪定(DEX)对糜烂性毒剂2-氯乙基乙基硫醚(CEES)诱导的人支气管上皮细胞损伤的潜在保护作用及机制。方法:使用浓度为1.2 mmol/L的CEES处理BEAS-2B细胞24 h,构建糜烂性毒剂导致细胞损伤的模型。使用不同浓度(0.1~1000μmol/L)的DEX预处理BEAS-2B细胞2 h后加CEES染毒24 h,再用CCK-8法检测细胞活力。选择DEX最佳保护剂量,在光镜下观察细胞形态,使用Annexin V/PI双染法检测细胞凋亡率,DHE、MitoSOX和RHOD123荧光探针分别检测细胞内总活性氧(ROS)、线粒体ROS(mtROS)和线粒体膜电位水平,通过试剂盒检测总超氧化物歧化酶(SOD)、CuZn-SOD、Mn-SOD酶活性和谷胱甘肽(GSH)含量变化。结果:与对照组相比,CEES染毒组细胞活力均明显降低(P<0.01),凋亡细胞数显著升高(P<0.01),细胞内总ROS、mtROS水平均明显升高(P<0.01),线粒体膜电位显著降低(P<0.01),总SOD、CuZn-SOD、Mn-SOD酶活性均升高(P<0.01),GSH含量降低(P<0.01)。与CEES染毒组相比,0.1~1000μmol/L DEX干预组细胞活力均升高(P<0.01),其中10μmol/L DEX干预组细胞活力升高最明显(P<0.01),后续用10μmol/L作为DEX干预组的最佳剂量。使用10μmol/L DEX干预显著抑制了CEES诱导的细胞凋亡(P<0.01),DEX干预组ROS和mtROS均降低(P<0.01),线粒体膜电位升高(P<0.05),总SOD、CuZn-SOD酶活力升高(P<0.01),GSH含量升高(P<0.01)。结论:DEX对CEES导致的人支气管上皮细胞损伤具有保护作用,其机制可能与调控线粒体稳态、降低mtROS的产生及增强细胞抗氧化水平有关。OBJECTIVE:To investigate protective effect and mechanism of dexmedetomidine hydrochloride(DEX)on blister agent 2-chloroethyl ethyl sulfide(CEES)-induced injury in human bronchial epithelial cells.METHODS:BEAS-2B cells were exposed to 1.2 mmol/L CEES for 24 h to establish a cell damage model induced by blister agents.The cells were pretreated with various concentrations of DEX for 2 h and exposed to CEES for 24 h.Cell viability was detected by the CCK-8 method to determine the optimal protective dose of DEX.Cell morphology was observed under a light microscope,and cell apoptosis rates were measured using Annexin V/PI staining.DHE,MitoSOX and RHOD123 fluorescent probes were employed to detect total reactive oxygen species(ROS),mitochondrial ROS(mtROS)and mitochondrial membrane potential levels,respectively.Enzyme activities of total SOD,CuZn-SOD and Mn-SOD and content of GSH were detected by kits.RESULTS:Compared with the control group,the CEES group showed a significant decrease in cell viability(P<0.01),a significant increase in cell apoptosis(P<0.01),a significant increase in ROS and mtROS(P<0.01),a significant decrease in mitochondrial membrane potential(P<0.01),an increase in total SOD,CuZn-SOD and Mn-SOD enzyme activity(P<0.01),and a decrease in GSH content(P<0.01).Compared with the CEES group,the cell viability of 0.1-1000μmol/L DEX intervention group was increased,and the most significant increase in cell viability was observed in the 10μmol/L DEX intervention group(P<0.01),and 10μmol/L was used as the optimal dose for the follow DEX intervention group.Apoptosis was decreased after the 10μmol/L DEX intervention(P<0.01).Compared with the CEES treatment group,DEX intervention reduced mtROS(P<0.01),increased mitochondrial membrane potential(P<0.05),increased enzyme activity of total SOD and CuZn-SOD(P<0.01),and increased GSH content(P<0.01).CONCLUSION:DEX exerted a protective effect on CEES-induced injury in human bronchial epithelial cells,and its mechanism could be related to the regulation of mitochondr

关 键 词：糜烂性毒剂 盐酸右美托咪定 线粒体 氧化应激 活性氧 

分 类 号：R858.5[医药卫生—航空、航天与航海医学]