miR-181a参与调控香烟提取物诱导的NR8383肺泡巨噬细胞自噬紊乱与促炎因子的生成  

miR-181a regulates CSE-induced autophagy dysfunction and releases of pro-inflammatory factors in NR8383 alveolar macrophages

作　　者：粱珍珍 吕燕平[1] 张艳莉[1] 解玉东 韩丽丽[1] LIANG Zhen-zhen;Lü Yan-ping;ZHANG Yan-li;XIE Yu-dong;HAN Li-li(Department of Respiratory Medicine, Zhoukou Central Hospital, Zhoukou 466000, China)

机构地区：[1]周口市中心医院呼吸科,河南周口466000

出　　处：《中国病理生理杂志》2019年第4期710-717,共8页Chinese Journal of Pathophysiology

基　　金：河南省医学科技攻关计划项目(No.201401011)

摘　　要：目的:探讨微小RNA-181a(miR-181a)对香烟提取物(CSE)诱导的NR8383大鼠肺泡巨噬细胞自噬紊乱与促炎因子生成的影响。方法:采用5%、10%和20%浓度的CSE刺激NR8383细胞,ELISA法检测肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和IL-8的分泌,RT-qPCR检测miR-181a水平,Cyto-ID染色检测自噬体数量,Western blot法检测LC3-Ⅱ、beclin-1和p62的表达。在20%CSE条件下,采用自噬抑制剂3-甲基腺嘌呤(3-MA)或自噬激动剂雷帕霉素(Rapa)预处理细胞,ELISA检测TNF-α、IL-6和IL-8的分泌;进一步转染miR-181a mimic或miR-181a inhibitor后,分别采用ELISA和Western blot观察在20%CSE条件下,细胞TNF-α、IL-6和IL-8分泌及LC3-Ⅱ、beclin-1和p62表达的情况。结果:CSE浓度依赖性促进NR8383细胞促炎因子生成和自噬紊乱;3-MA促进CSE诱导的NR8383细胞促炎因子释放,而Rapa部分逆转CSE诱导的NR8383细胞促炎因子释放;miR-181a mimic显著抑制CSE诱导的NR8383细胞促炎因子生成,促进自噬,miR-181a inhibitor促进CSE诱导的NR8383细胞促炎因子生成,加剧自噬紊乱。结论:miR-181a调控CSE诱导的NR8383细胞促炎因子释放可能与其调控自噬紊乱有关。AIM: To investigate the effect of microRNA-181 a(miR-181 a) on cigarette smoke extract(CSE)-induced autophagy disorder and releases of pro-inflammatory factors in NR8383 rat alveolar macrophages. METHODS: The NR8383 cells were treatment with 5%,10% and 20% CSE. The release levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) and IL-8 were measured by ELISA. The level of miR-181 a was detected by RT-qPCR. The numbers of autophagosomes were observed by Cyto-ID staining. The expression levels of LC3-Ⅱ, beclin-1 and p62 were determined by Western blot. NR8383 cells were pretreated with autophagy inhibitor 3-methyladenine(3-MA) or autophagy agonist rapamycin(Rapa) before treatment with 20% CSE, and the release levels of TNF-α, IL-6 and IL-8 were measured by ELISA. Furthermore, NR8383 cells were transfected with miR-181 a mimic or miR-181 a inhibitor before treatment with 20% CSE, and the release levels of TNF-α, IL-6 and IL-8, and the expression of LC3-Ⅱ, beclin-1 and p62 were detected by ELISA and Western blot, respectively. RESULTS: CSE increased release levels of pro-inflammatory factors and autophagy disorder in a concentration-dependent manner in the NR8383 cells(P<0.05). 3-MA increased CSE-induced releases of pro-inflammatory factors. However, Rapa partially reversed CSE-induced releases of pro-inflammatory factors. Additionally, miR-181 a mimic inhibited CSE-induced releases of pro-inflammatory factors and promoted autophagy. However, miR-181 a inhibitor increased CSE-induced releases of pro-inflammatory factors and autophagy disorder. CONCLUSION: miR-181 a regulates CSE-induced releases of pro-inflammatory factor in the NR8383 cells, which may be related to the regulatory role of miR-181 a in autophagy disorder.

关 键 词：微小RNA-181a 肺泡巨噬细胞 慢性阻塞性肺疾病 炎症 自噬 

分 类 号：R563[医药卫生—呼吸系统] R363.2[医药卫生—内科学]