电子烟急性暴露对小鼠BALF及肺表面活性蛋白的影响研究  

Effect of acute exposure to electronic cigarette on bronchoalveolar lavage fluid and pulmonary surfactant protein in mice

作　　者：王寅丹 李婷[1] 张国庆 章璐 张进娜 胡人杰 刘翠清[1] WANG Yindan;LI Ting;ZHANG Guoqing;ZHANG Lu;ZHANG Jinna;HU Renjie;LIU Cuiqing(School of Public Health,Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310056,China)

机构地区：[1]浙江中医药大学公共卫生学院,浙江杭州310056

出　　处：《预防医学》2022年第5期456-460,共5页CHINA PREVENTIVE MEDICINE JOURNAL

摘　　要：目的探索急性暴露于电子烟的小鼠支气管肺泡灌洗液(BALF)白细胞计数、总蛋白含量以及肺表面活性蛋白水平,为电子烟的呼吸系统损害作用机制研究提供实验依据。方法21只C57BL/6N雌鼠随机分为空白对照组、溶剂对照组和尼古丁组。溶剂对照组和尼古丁组小鼠每天分别暴露于电子烟烟液载体溶剂气溶胶和含25 mg/mL尼古丁的电子烟气溶胶3 h,空白对照组饲养在洁净空气中。暴露3 d后收集小鼠BALF,通过瑞氏-姬姆萨染色镜检观察细胞形态,统计白细胞数量,采用BCA法检测总蛋白含量;取小鼠肺组织采用实时荧光定量PCR检测表面活性蛋白基因mRNA表达水平。结果3组小鼠生长状况良好,无明显异常,无死亡。尼古丁组小鼠BALF含大量单核-巨噬细胞,空白对照组和溶剂对照组相对较少。空白对照组、溶剂对照组和尼古丁组小鼠BALF中白细胞数分别为(2.00±0.77)×10^(7)个/L、(1.79±0.99)×10^(7)个/L和(4.00±1.35)×10^(7)个/L,总蛋白含量分别为(0.16±0.03)、(0.12±0.02)、(0.16±0.04)mg/mL,肺表面活性蛋白B(SP-B)mRNA相对表达量分别为1.00±0.14、0.82±0.12和0.74±0.07,SP-D mRNA相对表达量分别为1.00±0.06、0.90±0.02和0.71±0.15,3组比较差异均有统计学意义(F=9.199,P=0.002;F=3.610,P=0.048;F=5.491,P=0.028;F=10.460,P=0.005)。尼古丁组小鼠白细胞数高于空白对照组和溶剂对照组小鼠(P=0.007、0.003),总蛋白含量高于溶剂对照组小鼠(P=0.060),SP-B mRNA相对表达量低于空白对照组小鼠(P=0.025),SP-D mRNA相对表达量低于空白对照组和溶剂对照组小鼠(P=0.004、0.041)。结论电子烟急性暴露引起小鼠肺内炎症水平升高,肺部毛细血管屏障受损并降低肺表面活性蛋白的表达。Objective To assess the effects of acute exposure to electronic cigarette(e-cigarette)on leukocyte and total protein levels in bronchoalveolar lavage fluid(BALF)and pulmonary surfactant protein expression in a mouse model,so as to provide insights into the elucidation of the mechanism underlying the damages to the respiratory system caused by e-cigarette.Methods Twenty-one C57BL/6N female mice were randomly divided into the blank control group,the sol⁃vent control group and the nicotine group.Mice in the solvent control group and the nicotine group were exposed to the solvent aerosol or e-cigarette aerosol containing 25 mg/mL nicotine for 3 hours daily,while mice in the blank con⁃trol group were bred in clean air.Following 3-day exposure,mouse BALF and lung specimens were collected.The cell morphology was observed using microscopy following Wright-Giemsa staining and the leukocyte count was estimated in BALF,while the total protein expression was quantified using bicinchoninic acid(BCA)assay.In addition,the mRNA expression of pulmonary surfactant protein genes was detected in mouse lung specimens using quantitative real-time PCR(qPCR)assay.Results All mice in three groups grew well without obvious abnormality or death seen.Wright-Gi⁃emsa staining showed a higher number of mononuclear macrophages in mouse BALF in the nicotine group than in the blank control group and the solvent control group.The leukocyte counts were(2.00±0.77)×10^(7),(1.79±0.99)×10^(7) and(4.00±1.35)×10^(7) cells/L(F=9.199,P=0.002),and the total protein levels were(0.16±0.03),(0.12±0.02)and(0.16±0.04)mg/mL in mouse BALF in the blank control group,solvent control group and nicotine group(F=3.610,P=0.048),and the relative mRNA expression of pulmonary surfactant protein B(SP-B)and SP-D was 1.00±0.14,0.82±0.12 and 0.74±0.07(F=5.491,P=0.028),and 1.00±0.06,0.90±0.02 and 0.71±0.15 in mouse lung specimens,respectively(F=10.460,P=0.005).The leukocyte count was significantly higher in the nicotine group than in the blank control group

关 键 词：电子烟 尼古丁 白细胞 支气管肺泡灌洗液 肺表面活性蛋白 

分 类 号：R361.3[医药卫生—病理学]