二氧化硅所致急性肺损伤大鼠肺部菌群的特征及相关性分析  

Characterization and correlation analysis of lung flora in rats with silica-induced acute lung injury

作　　者：刘畅 芦俊 肖荣 李迎秋[1] 胡珏 田玥 张家祥 卢芳国[1] LIU Chang;LU Jun;XIAO Rong;LI Yingqiu;HU Jue;TIAN Yue;ZHANG Jiaxiang;LU Fangguo(Medical School,Hunan University of Chinese Medicine,Changsha 410208,China;School of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Provincial Hospital of Integrated Traditional Chinese and Western Medicine,Changsha 410006,China)

机构地区：[1]湖南中医药大学医学院,湖南长沙410208 [2]湖南中医药大学中西医结合学院,湖南长沙410208 [3]湖南省中西医结合医院,湖南长沙410006

出　　处：《中国病理生理杂志》2024年第1期81-88,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82374266,No.82074250);湖南省教育厅优青项目(No.23B0342);湖南省大学生创新创业训练项目(No.S202210541126);湖南中医药大学科研项目(No.2021XJJJ005);湖南中医药大学本科生科研创新基金资助项目(No.2023BKS058)。

摘　　要：目的:探讨二氧化硅所致急性肺损伤大鼠模型的潜在生物学机制。方法:采用随机数字表法,将SD雄性大鼠划分为正常组和二氧化硅模型组。正常组经气管灌注1 mL生理盐水,模型组经气管灌注等体积50 g/L二氧化硅混悬液。7 d后收集大鼠肺组织和血清,HE染色观察肺组织病理学特征,免疫组化检测肺组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)和消皮素D(GSDMD)蛋白表达,ELISA法检测血清炎症因子白细胞介素1β(IL-1β)、IL-18和肿瘤坏死因子α(TNF-α)水平;从肺组织中提取细菌DNA,经16S核糖体RNA基因测序描述肺部菌群组成变化,通过生物信息学分析比较正常组与模型组菌群结构差异。结果:与正常组相比,模型组大鼠体重增长率和肺指数有显著差异;模型组大鼠肺组织HE染色可见肺泡结构破坏,炎性细胞增多;模型组大鼠肺组织NLRP3和GSDMD蛋白表达水平均显著升高,血清IL-1β、IL-18和TNF-α水平均显著升高。与正常组相比,模型组丰度增加的肺部菌群为双歧杆菌(Bifidobacterium)、Clostridium sensu stricto 1和副萨特氏菌(Parasutterella)。Spearman相关性分析显示,肺组织NLRP3/GSDMD和血清IL-1β/TNF-α水平与Parasutterella丰度呈正相关。结论:二氧化硅所致急性肺损伤的病理机制可能与肺部菌群结构差异和炎症水平升高有关。因此,调节肺部菌群可能为防治二氧化硅引起的急性肺损伤提供新的思路。AIM:To elucidate the possible biological mechanism of silica-induced acute lung injury in rats.METHODS:Sixteen Male Sprague-Dawley rats were divided into control and acute silicosis model groups,and instilled intratracheally with 1 mL of normal saline and 50 g/L silica suspension,respectively.After 7 d,the rats were sacrificed for collection of lung tissue and serum.The serum levels of interleukin-1β(IL-1β),IL-18 and tumor necrosis factor-α(TNF-α)were measured by using ELISA.The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and gasdermin D(GSDMD)were measured by immunohistochemistry.Bacterial DNA was extracted from the lung tissue for 16S ribosomal RNA gene sequencing to characterize changes in the composition of lung flora.The differences in the structure of bacterial flora between control and model groups were analyzed by bioinformatic analyses.RESULTS:Immunohistochemical analysis showed that the protein expression levels of NLRP3 and GSDMD were higher in the lungs of the rats in model group.In addition,serum cytokine profiling showed that IL-1β,IL-18 and TNF-αlevels were significantly higher in model group.The most abundant bacterial genera in the lung flora of the rats in model group were Bifidobacterium,Clostridium sensu stricto 1,and Parasutterella.The NLRP3 and GSDMD levels in the lung tissue and IL-1βand TNF-αlevels in serum were positively correlated with the abundance of Parasutterella.CONCLUSION:The alterations in lung flora structure and increased inflammation levels may be the actual biological mechanisms underlying silica-induced acute lung injury.The modulation of lung flora may provide a basis for the prevention and treatment of silica-induced acute lung injury.

关 键 词：矽肺 二氧化硅 急性肺损伤 肺部菌群 

分 类 号：R135.2[医药卫生—劳动卫生] R563[医药卫生—公共卫生与预防医学] R363.2