松木屑烟雾溶液对大鼠肺泡Ⅱ型上皮细胞损伤的研究  

Effect of Pine Sawdust Smoke Solution Induced Injury in Rat Alveolar Type I Epithelial Cells

作　　者：崔培 陈安宁 詹球 李榕生 朱秀连 蒙凤姬 朱富军 杨福旺 童亚林 辛海明 CUI Pei;CHEN Anning;ZHAN Qiu;LI Rongsheng;ZHU Xiulian;MENG Fengji;ZHU Fujun;YANG Fuwang;TONG Yalin;XIN Haiming(Animal Laboratory,NO.924 Hospital of the Joint Logistics Support Force,Guilin Guangri 541002,China)

机构地区：[1]联勤保障部队924医院动物实验室,广西桂林541002 [2]联勤保障部队924医院烧伤整形皮肤美容中心,广西桂林541002

出　　处：《联勤军事医学》2024年第10期824-831,共8页Military Medicine of Joint Logistics

基　　金：广西青年科学基金项目(2024GXNSFBA010044);广西壮族自治区临床重点专科建设项目(桂卫医发[2023]26号);联勤保障部队第924医院科技计划项目(GS2020FH08、GS2020CZ05、GS2020CZ06)。

摘　　要：目的探讨不同浓度松木屑烟雾溶液对大鼠肺泡Ⅱ型上皮细胞(alveolar typeⅡepithelial cell,AT-Ⅱ)活性、细胞凋亡和炎性细胞因子表达的影响。方法将松木屑燃烧产生的烟雾通过负压排气法收集到二甲基亚砜溶液(dimethyl sulfoxide,DMSO)中并混匀(以下简称为烟雾溶液)。根据与AT-Ⅱ共培养的干预因素不同,分为正常组(AT-Ⅱ细胞未经任何处理)、对照组(细胞培养基中加入1.00‰DMSO)、低浓度致伤组(细胞培养基中加入0.50‰烟雾溶液)、中浓度致伤组(细胞培养基中加入0.75‰烟雾溶液)和高浓度致伤组(细胞培养基中加入1.00‰烟雾溶液),致伤后6、12、24和48 h,采用细胞计数试剂盒(cell counting kit 8,CCK-8)检测AT-Ⅱ活性,流式细胞术检测AT-Ⅱ凋亡情况,酶联免疫吸附分析(enzyme-linked immunosorbent assay,ELISA)检测细胞匀浆上清液中白细胞介素6(interleukin 6,IL-6)和肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)的含量。结果致伤后不同时间点、不同组别间细胞增殖、细胞凋亡变化差异均有统计学意义(P均<0.001),低、中、高浓度致伤组细胞增殖情况随着烟雾浓度的增加逐渐降低,细胞凋亡率随着烟雾浓度的增加逐渐升高(P均<0.05)。致伤后不同时间点、不同组别间细胞中IL-6、TNF-α水平差异均有统计学意义(P均<0.001),且烟雾浓度越高,细胞中IL-6、TNF-α水平越高。结论低、中、高浓度的松木屑烟雾溶液均可显著降低AT-Ⅱ增殖活性,促使细胞凋亡,增加IL-6和TNF-α的表达。Objective To investigate the effects of different concentrations of pine sawdust smoke solution on the activity,apoptosis and inflammatory cytokine expression of alveolar typeⅡepithelial cells(AT-Ⅱ)in rats.Methods Smoke produced by burning pine sawdust was collected into dimethyl sulfoxide(DMSO)solution by negative pressure suction and mixed(smoke solution for short).According to the different intervention factors of co-culture with AT-Ⅱ,the experiment was divided into normal group(AT-Ⅱcells were not treated),control group(culture with 1.00‰DMSO),low concentration injury group(culture with 0.50‰smoke solution),medium concentration injury group(culture with 0.75‰smoke solution),and high concentration injury group(culture with 1.00‰smoke solution),at 6 hours,12 hours,24 hours and 48 hours after injury,the activity of AT-Ⅱwas detected by cell counting kit 8(CCK-8)method,the apoptosis of AT-Ⅱwas detected by flow cytometry,and the contents of interleukin 6(IL-6)and tumor necrosis factor alpha(TNF-α)in the supernatant of cell homogenate were detected by enzyme-linked immunosorbent assay(ELISA).Results At different time points after injury,there were statistically significant differences in cell proliferation and apoptosis among different groups(all P<0.001),cell proliferation in low,medium and high concentration injury groups gradually decreased with the increase of smog concentration,and cell apoptosis rate gradually increased with the increase of smog concentration(all P<0.05).At different time points after injury,the levels of IL-6 and TNF-αin cells of different groups had statistical significance(all P<0.001),and the higher the concentration of smoke,the higher the levels of IL-6 and TNF-αin cells.Conclusion Low,medium and high concentrations of pine sawdust smoke solution could significantly reduce the proliferative activity of AT-Ⅱ,promote cell apoptosis,and increase the expression of IL-6 and TNF-α.

关 键 词：松木屑烟雾 吸入性肺损伤 肺泡Ⅱ型上皮细胞 细胞活性 细胞凋亡 炎症 

分 类 号：R563[医药卫生—呼吸系统]