摘要
目的探讨干扰素λ1(interferon-λ1,IFN-λ1)对呼吸道合胞病毒(respiratory syncytial virus,RSV)刺激下的人支气管上皮细胞(human bronchial epithelial cell,HBEC)糖皮质激素(glucocorticoid,GC)抵抗的影响。方法将HBEC分为对照组、地塞米松(dexamethasone,Dex)组、IFN-λ1组、RSV组、RSV+IFN-λ1组。采用CCK-8法测定不同浓度IFN-λ1对HBEC细胞活力的影响及各组HBEC对地塞米松的敏感性,实时荧光定量PCR法测定p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)、糖皮质激素受体(glucocorticoid receptor,GR)、MAPK磷酸酶-1(MAPK phosphatase-1,MKP-1)mRNA表达,Western blot法测定细胞核、细胞质GR蛋白表达,并计算细胞核GR蛋白量/细胞质GR蛋白量比值。结果24 h、72 h时,随IFN-λ1浓度增加,HBEC细胞增殖活性增加,且呈浓度和时间依赖性(P<0.05);与RSV组比较,RSV+IFN-λ1组地塞米松的半数最大抑制浓度、p38 MAPK mRNA表达水平降低(P<0.05),GR、MKP-1 mRNA表达水平、细胞核与细胞质GR水平及细胞核GR/细胞质GR比值升高(P<0.05)。结论IFN-λ1可通过上调MKP-1抑制p38 MAPK通路,促进GR的核转移,从而改善HBEC细胞GC抵抗。
Objective To investigate the effect of interferon-λ1(IFN-λ1)on glucocorticoid(GC)resistance in human bronchial epithelial cells(HBECs)stimulated by respiratory syncytial virus(RSV).Methods HBECs were divided into five groups:control,dexamethasone,IFN-λ1,RSV,and RSV+IFN-λ1.CCK-8 assay was used to measure the effect of different concentrations of IFN-λ1 on the viability of HBECs,and the sensitivity of HBECs to dexamethasone was measured in each group.Quantitative real-time PCR was used to measure the mRNA expression levels of p38 mitogen-activated protein kinase(p38 MAPK),glucocorticoid receptor(GR),and MAPK phosphatase-1(MKP-1).Western blot was used to measure the protein expression level of GR in cell nucleus and cytoplasm,and the nuclear/cytoplasmic ratio of GR was calculated.Results At 24 and 72 hours,the proliferation activity of HBECs increased with the increase in IFN-λ1 concentration in a dose-and time-dependent manner(P˂0.05).Compared with the RSV group,the RSV+IFN-λ1 group had significant reductions in the half-maximal inhibitory concentration of dexamethasone and the mRNA expression level of p38 MAPK(P<0.05),as well as significant increases in the mRNA expression levels of GR and MKP-1,the level of GR in cell nucleus and cytoplasm,and the nuclear/cytoplasmic GR ratio(P<0.05).Conclusions IFN-λ1 can inhibit the p38 MAPK pathway by upregulating MKP-1,promote the nuclear translocation of GR,and thus ameliorate GC resistance in HBECs.
作者
彭力
刘瑶
李芳彩
丁小芳
林小娟
杨图宏
钟礼立
PENG Li;LIU Yao;LI Fang-Cai;DING Xiao-Fang;LIN Xiao-Juan;YANG Tu-Hong;ZHONG Li-Li(Children's Medical Center,Hunan Provincial People's Hospital/First Affiliated Hospital of Hunan Normal University/Hunan Key Laboratory of Pediatric Respiratory Diseases,Changsha 410005,China)
出处
《中国当代儿科杂志》
北大核心
2025年第8期1011-1016,共6页
Chinese Journal of Contemporary Pediatrics
基金
湖南省人民医院仁术基金项目(RS2022B03)
湖南省卫生健康委国家临床重点专科重大科研专项项目(Z2023054)。
关键词
糖皮质激素抵抗
干扰素λ1
呼吸道合胞病毒
p38
MAPK
人支气管上皮细胞
Interferon-λ1
Respiratory syncytial virus
Glucocorticoid resistance
p38 mitogen-activated protein kinase
Human bronchial epithelial cell
