氢吗啡酮对脂多糖诱导小鼠肺泡巨噬细胞高尔基体应激影响  

Effect of hydromorphone on Golgi stress in lipopolysaccharide-attacked mouse alveolar macrophages

作　　者：李少娜 贾长新[1] 吴秀云[1] 朱尤壮 赵芹[1] 徐业翔 LI Shaona;JIA Changxin;WU Xiuyun;ZHU Youzhuang;ZHAO Qin;XU Yexiang(Department of Anesthesiology,The Affiliated Hospital of Qingdao University,Qingdao 266000,China)

机构地区：[1]青岛大学附属医院麻醉科,山东青岛266000 [2]青岛大学附属医院急诊内科,山东青岛266000

出　　处：《青岛大学学报（医学版）》2024年第4期523-527,共5页Journal of Qingdao University(Medical Sciences)

基　　金：山东省医药卫生技术发展计划项目(20210411-0960);青岛大学附属医院青年科研基金(QDFYQN2023227)。

摘　　要：目的探讨氢吗啡酮(HM)对脂多糖(LPS)诱导的小鼠肺泡巨噬细胞高尔基体应激的影响及其机制。方法以10mg/LLPS处理小鼠肺泡巨噬细胞系MH-S,构建肺泡巨噬细胞高尔基体应激模型。将细胞随机分为Control组(A组,正常培养)、LPS组(B组,给予10mg/L的LPS)、LPS+HM组(C组,给予1μmol/LHM和10mg/LLPS)、Nrf2siRNA+LPS+HM组(D组,Nrf2siRNA转染后给予1μmol/LHM和10mg/LLPS)和NC siRNA+LPS+HM组(E组,NCsiRNA转染后给予1μmol/LHM和10mg/LLPS)。采用ELISA法检测细胞上清液白细胞介素1β(IL-1β)和白细胞介素6(IL-6)含量;检测丙二醛(MDA)含量与超氧化物歧化酶(SOD)活性;采用Westernblot法检测核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、高尔基体基质蛋白130(GM130)、高尔基体蛋白97(Golgin-97)、甘露糖苷酶Ⅱ(MannosidaseⅡ)、钙转运ATP酶2C型成员1(ATP2C1)蛋白表达水平;透射电镜下观察细胞高尔基体形态学改变。结果与A组比较,B组IL-1β、IL-6、MDA含量显著升高及Nrf2、HO-1蛋白表达上调,SOD活性显著降低及GM130、Golgin-97、MannosidaseⅡ、ATP2C1蛋白的表达下调(F=2.33~47.61,P<0.05);与B组比较,C组IL-1β、IL-6、MDA含量显著降低,SOD活性显著升高及Nrf2、HO-1、GM130、Golgin-97、MannosidaseⅡ、ATP2C1蛋白表达上调(P<0.05);与C组比较,D组IL-1β、IL-6、MDA含量显著升高,SOD活性显著降低及Nrf2、HO-1、GM130、Golgin-97、MannosidaseⅡ、ATP2C1蛋白表达下调(P<0.05)。透射电镜显示,C组细胞高尔基体损伤、空泡化与碎片化较B组减轻;D组细胞高尔基体损伤、空泡化与碎片化较C组加重。结论HM通过调控Nrf2/HO-1信号通路抑制LPS诱导的肺泡巨噬细胞高尔基体应激,减轻细胞炎症与氧化应激反应。Objective To investigate the effect of hydromorphone(HM)on Golgi stress in lipopolysaccharide(LPS)-attacked mouse alveolar macrophages and its mechanism.Methods The mouse alveolar macrophage cell line MH-S was attacked with 10 mg/L LPS to establish a model of Golgi stress,and then the cells were randomly divided into control group(group A,normal culture),LPS group(group B treated with 10 mg/L LPS),LPS+HM group(group C treated with 1μmol/L HM and 10 mg/L LPS),Nrf2 siRNA+LPS+HM group(group D treated with 1μmol/L HM and 10 mg/L LPS after Nrf2 siRNA transfection),and NC siRNA+LPS+HM group(group E treated with 1μmol/L HM and 10 mg/L LPS after NC siRNA transfection).ELISA was used to measure the content of interleukin 1β(IL-1β)and interleukin 6(IL-6)in cell supernatant;the content of malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)were measured;Western blot was used to measure the protein expression levels of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),Golgi Matrix Protein of 130(GM130),Golgin-97,MannosidaseⅡ,and ATP2C1;transmission electron microscopy was used to observe the morphological changes of Golgi apparatus.Results Compared with group A,group B had significant increases in the content of IL-1β,IL-6,and MDA and the protein expression levels of Nrf2 and HO-1 and significant reductions in the activity of SOD and the protein expression levels of GM130,Golgin-97,MannosidaseⅡ,and ATP2C1(F=2.33-47.61,P<0.05).Compared with group B,group C had significant reductions in the content of IL-1β,IL-6,and MDA and significant increases in the activity of SOD and the protein expression levels of Nrf2,HO-1,GM130,Golgin-97,MannosidaseⅡ,and ATP2C1(P<0.05).Compared with group C,group D had significant increases in the content of IL-1β,IL-6,and MDA and significant reductions in the activity of SOD and the protein expression levels of Nrf2,HO-1,GM130,Golgin-97,MannosidaseⅡ,and ATP2C1(P<0.05).Transmission electron microscopy showed that compared with group B,group C had all

关 键 词：氢吗啡酮 脂多糖类 巨噬细胞 肺泡 高尔基体 氧化性应激 血红素加氧酶-1 小鼠 

分 类 号：R965[医药卫生—药理学]