二氧化硅染尘小鼠肺纤维化相关炎症和氧化应激因子的响应及关键基因表达  

Responses of inflammatory factors and oxidative stress factors and expressions of key genes associated with pulmonary fibrosis in silica-exposed mice

作　　者：吕达 杨慧楠 刘春城 王乐 赵宏宇 蔡禄[1,2] LYU Da;YANG Huinan;LIU Chuncheng;WANG Le;ZHAO Hongyu;CAI Lu(School of Life Science and Technology,Inner Mongolia University of Science and Technology,Baotou,Inner Mongolia 014010,China;Inner Mongolia Autonomous Region Key Laboratory of Functional Genome Bioinformatics,Inner Mongolia University of Science and Technology,Baotou,Inner Mongolia 014010,China)

机构地区：[1]内蒙古科技大学生命科学与技术学院,内蒙古包头014010 [2]内蒙古科技大学内蒙古自治区功能基因组生物信息学重点实验室,内蒙古包头014010

出　　处：《环境与职业医学》2021年第10期1150-1155,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金(62071259,61671256,31760247);内蒙古自然科学基金(2021MS03007)。

摘　　要：[背景]二氧化硅(SiO_(2))长期沉积在肺组织中能够诱发炎症与纤维化反应,进而导致尘肺病,但其发病机制尚未明确。[目的]探究SiO_(2)对小鼠肺部纤维化的影响及其相关机制。[方法]将35只C57BL/6小鼠随机分成空白对照组、生理盐水对照组、20 g·L^(-1) SiO_(2)染尘组、60 g·L^(-1) SiO_(2)染尘组4个组别,其中空白对照组5只,其余组别各10只。采用非气管暴露法在第0天和第14天进行2次染尘,按照小鼠体重灌注不同体积的SiO_(2)悬浊液。在染尘第28天和第56天时取小鼠肺组织和血清。10倍光学显微镜下观察肺组织HE切片;ELISA法检测肺组织中小鼠成纤维生长因子(FGF)、肿瘤坏死因子-α(TNF-α)和转化生长因子-β(TGF-β)3种炎症因子的质量分数(后简称含量),检测血清中超氧化物歧化酶(SOD)、活性氧(ROS)、丙二醛(MDA)3种氧化应激因子的水平;10倍显微镜下观察Masson染色切片胶原纤维分布;分光光度计检测肺组织中羟脯氨酸含量;实时荧光定量PCR技术检测肺组织中Col1a2和α-SMA mRNA表达水平的变化。[结果]HE染色结果显示:SiO_(2)染尘组与生理盐水对照组相比有明显的炎症细胞浸润,部分出现纤维结节。炎症因子检测结果显示:20 g·L^(-1)和60 g·L^(-1)染尘组小鼠在染尘第56天时,TGF-β、FGF、TNF-α含量分别是3.42、3.62 ng·g^(-1),15.20、19.78 ng·g^(-1)和9.02、11.30 ng·g^(-1),较生理盐水组均有增高,差异有统计学意义(P<0.01)。氧化应激因子检测结果显示:小鼠20 g·L^(-1)染尘组在染尘第28和56天时,MDA浓度分别是0.23、0.26μmol·L^(-1),ROS水平均为11 U·L^(-1),SOD质量浓度分别0.43、0.44μg·L^(-1);小鼠60 g·L^(-1)染尘组在染尘第28和56天时,MDA浓度为0.28、0.34μmol·L^(-1),ROS水平分别是13、14 U·L^(-1),SOD质量浓度分别0.38、0.36μg·L^(-1);相比于生理盐水对照组,在两个时间点两个浓度染尘组小鼠血清中MDA、ROS水平增加,SOD水平下降,差异均有�[Background]It is well known that long-term deposition of silicon dioxide(SiO_(2))in lung tissues can induce inflammation and fibrosis and result in pneumoconiosis,but pathogenesis of pneumoconiosis remains elusive.[Objective]This experiment investigates the effect of SiO_(2) on pulmonary fibrosis in mice.[Methods]Thirty-five C57BL/6 mice were randomly divided into four groups:a blank control group(5 mice),a normal saline control group(10 mice),a 20 g·L^(-1) SiO_(2) group(10 mice),and a 60 g·L^(-1) SiO_(2) group(10 mice).The mice were exposed by intratracheal instillation of SiO_(2) suspension on day 0 and day 14,and the lung tissue and serum samples were collected on day 28 and day 56 after silica or saline instillation.HE sections of lung tissues were observed under optical microscope(10×magnification).ELISA was used to detect the levels of three inflammatory cytokines including fibroblast growth factor(FGF),tumor necrosis factor-α(TNF-α),and transforming growth factor-β(TGF-β)in lung tissues,and three oxidative stress factors including superoxide dismutase(SOD),reactive oxygen species(ROS),and malondialdehyde(MDA)in serum.The distributions of collagen fibers in Masson-stained sections were observed under microscope(10×magnification).The level of hydroxyproline(HYP)in lung tissues was detected with a spectrophotometer.The expressions of Col1a2 andα-SMA mRNA in lung tissues were measured by real-time PCR.[Results]The results of HE staining showed that there were obvious infiltration of inflammatory cells and some fibrous nodules in the two SiO_(2) groups compared with the normal saline control group.The detection results of inflammatory factors indicated that on day 56,the levels of TGF-βin lung tissues of the 20 g·L^(-1) SiO_(2) group and the 60 g·L^(-1) SiO_(2) group were 3.42 and 3.62 ng·g^(-1),the levels of FGF were 15.20 and 19.78 ng·g^(-1),the levels of TNF-αwere 9.02 and 11.30 ng·g^(-1),respectively,all higher than those of the normal saline group(P<0.01).The detection results of oxidative

关 键 词：肺纤维化 二氧化硅 胶原沉积 炎症反应 氧化应激 

分 类 号：R114[医药卫生—卫生毒理学]