维生素D抑制细颗粒物(PM_(2.5))诱导的A549人肺泡上皮细胞自噬及细胞因子释放  

Vitamin D inhibits PM_(2.5)-induced autophagy and cytokine release of A549 human alveolar epithelial cells

作　　者：黄婧 毛旭 丁朵 王蕾[3] HUANG Jing;MAO Xu;DING Duo;WANG Lei(Department of Rheumatism and Immunology,The First Affiliated Hospital,Xi’an Jiaotong University,Xi’an 710061;Department of Periodontology,Stomatological Hospital,Air Force Medical University,Xi’an 710032;Department of Pulmonary and Critical Care Medicine,The Second Affiliated Hospital,Xi’an Jiaotong University,Xi’an 710004,China)

机构地区：[1]西安交通大学第一附属医院风湿免疫科,陕西西安710061 [2]空军军医大学口腔医院牙周科,陕西西安710032 [3]西安交通大学第二附属医院呼吸与危重症科,陕西西安710004

出　　处：《细胞与分子免疫学杂志》2023年第1期9-14,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：陕西省自然科学基础研究计划(2022JQ-940);中国博士后科学基金会(2021M702610);中央高校基本科研业务费(xzy012020060)

摘　　要：目的探讨维生素D(VD)在细颗粒物(PM_(2.5))诱导A549人肺泡上皮细胞发生自噬过程中的作用及对上皮细胞因子产生的影响。方法燃烧器的乙炔扩散火焰上部采集制备PM_(2.5),采用PM_(2.5)体外处理A549细胞,并给予VD或自噬抑制剂3-甲基腺嘌呤(3-MA)处理。Western blot法检测自噬相关蛋白微管相关蛋白1轻链3Ⅱ/Ⅰ(LC3Ⅱ/Ⅰ)及beclin-1的表达,实时荧光定量PCR检测A549细胞白细胞介素25(IL-25)、IL-33及胸腺间质淋巴细胞生成素(TSLP)的mRNA表达,透射电镜观察自噬体的形成。结果PM_(2.5)导致A549细胞中LC3Ⅱ/Ⅰ比值、beclin-1蛋白表达增加,自噬体形成增多,给予VD处理后,PM_(2.5)诱导的细胞自噬水平降低;在3-MA抑制A549细胞自噬水平后,PM_(2.5)仍可诱导细胞自噬,且VD依然可逆转PM_(2.5)诱导的自噬;PM_(2.5)通过诱导自噬促进A549细胞分泌IL-25、IL-33及TSLP,而3-MA及VD可抑制该过程。结论VD抑制PM_(2.5)诱导的A549细胞自噬及细胞因子释放,发挥保护作用。Objective To investigate how vitamin D(VD)affects fine particulate matter(PM_(2.5))-induced autophagy and cytokines production in A549 human alveolar epithelial cells.Methods PM_(2.5)samples were prepared at the upper part of an acetylene diffusion flame burner.A549 cells were treated with PM_(2.5)in vitro,and/or were treated with VD or autophagy inhibitor 3-methyladenine(3-MA).Western blot analysis was employed to analyze the level of LC3-II/I and Beclin-1 in A549 cells with different groups.Real time quantitative PCR was used to detect the mRNA expression of interleukin-25(IL-25),IL-33 and thymic stromal lymphopoietin(TSLP).The formation of autophagosomes was observed by transmission electron microscopy.Results LC3-II/I ratio and beclin-1 protein expression were found increased in A549 cells after PM_(2.5)treatment,and autophagosome were increased too.There was a marked decrease of PM_(2.5)-induced autophagy with VD treatment.After 3-MA treatment,the autophagy was inhibited.Then,PM_(2.5)continued to induce autophagy,while VD could also reverse it.PM_(2.5)promoted the secretion of IL-25,IL-33 and TSLP by inducing autophagy in A549 cells while this process was inhibited by 3-MA and VD.Conclusion VD can inhibit PM_(2.5)-induced autophagy and cytokine release in A549 cells,thus playing a protective role.

关 键 词：A549细胞 肺泡上皮细胞 细颗粒物(PM_(2.5)) 自噬 免疫激活 维生素D(VD) 

分 类 号：R392.12[医药卫生—免疫学] R977.2[医药卫生—基础医学] R364.5