穿心莲内酯调控p38及ERK磷酸化改善甲型流感病毒诱导肺上皮细胞损伤的作用  

Andrographolide Improves Influenza A Virus Induced Lung Epithelial Cells Injury via Regulating p38 and ERK Phosphorylation

作　　者：李刚 田甜 王磊 徐淑娜 杨文平 LI Gang;TIAN Tian;WANG Lei;XU Shuna;YANG Wenping(Jinan People's Hospital Affiliated to Shandong First Medical University,Jinan 271119 China)

机构地区：[1]山东第一医科大学附属济南人民医院,济南271199

出　　处：《病毒学报》2024年第3期476-483,共8页Chinese Journal of Virology

摘　　要：甲型流感病毒(Influenza A virus,IAV)感染引起肺上皮细胞损伤与促进p38磷酸化、抑制细胞外调节蛋白激酶(Extracellular regulated protein kinases,ERK)磷酸化有关,穿心莲内酯(Andrographolide,AD)通过抗凋亡、抗炎、抗氧化等途径发挥细胞保护作用且对促分裂素原活化蛋白激酶38(Protein 38,p38)、ERK的磷酸化具有调控作用。为研究AD通过调控p38及ERK磷酸化改善IAV诱导肺上皮细胞损伤的作用,本文培养小鼠肺上皮细胞株MLE-12,按照MOI=0.2感染IAV 2h后给予不同剂量(6.25、12.5、25.0 mg/L)AD或25.0 mg/L AD联合对照溶剂(体积分数0.1%)、p38激动剂SB202190(10μmol/L)、ERK抑制剂PD98059(20μmol/L)处理10h,检测细胞的增殖抑制率、凋亡率,培养基中白介素(Interleukin,IL)-1β、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、IL-6、丙二醛(Malondialdehyde,MDA)、超氧化物歧化酶(Superoxide dismutase,SOD)、总抗氧化力(Total antioxidant capacity,T-AOC)的含量,细胞中p-p38、p-ERK的表达水平。结果显示:感染IAV的MLE-12细胞中SOD、T-AOC含量、pERK表达水平降低,增殖抑制率、凋亡率、IL-1β、TNF-α、IL-6、MDA含量及p-p38表达水平增加(P<0.05);不同剂量AD组MLE-12细胞中SOD、T-AOC含量、p-ERK表达水平增加,增殖抑制率、凋亡率、IL-1β、TNF-α、IL-6、MDA含量及p-p38表达水平降低(P<0.05);SB202190+AD组、PD98059+AD组MLE-12细胞中SOD、T-AOC含量降低,增殖抑制率、凋亡率、IL-1β、TNF-α、IL-6、MDA含量增加(P<0.05)。以上结果表明,AD通过抑制p38磷酸化、促进ERK磷酸化的方式抑制细胞凋亡、炎症反应及氧化应激反应,进而减轻IAV诱导肺上皮细胞损伤。The lung epithelial cells injury caused by Influenza A virus(IAV)infection is related to the promotion of p38 phosphorylation and the inhibition of extracellular regulated protein kinase(ERK)phosphorylation.Andrographolide(AD)plays cytoprotection roles through anti-apoptosis,anti-inflammatory,antioxidant and other ways,and has regulatory effects on the phosphorylation of p38 and ERK.To investigate the effect of AD on improving IAV induced lung epithelial cell injury via regulating p38 and ERK phosphorylation,mouse lung epithelial cell line MLE-12 was cultured.After infected with IAV at MOI=0.2 for 2 hours,different doses(6.25,12.5,25.0 mg/L)of AD or 25.0 mg/L AD combined with control solvent(volume fraction 0.1%),p38 agonist SB202190(10μmol/L),ERK inhibitor PD98059(20μmol/L)for 10 hours,the cell proliferation inhibitory rate,apoptosis rate,and the contents of interleukin-1(IL-1)β,tumor necrosis factor-α(TNF-α),IL-6,malondialdehyde(MDA),superoxide dismutase(SOD),total antioxidant capacity(T-AOC)in the culture medium,and the expression levels of p-p38 and p-ERK in cells were detected.The results showed that in MLE-12 cells infected with IAV,the contents of SOD,T-AOC,the expression level of p-ERK were significantly decreased,the proliferation inhibitory rate and apoptosis rate,the contents of IL-1β,TNF-α,IL-6,MDA,the expression level of p-p38 were significantly increased(P<0.05).In MLE-12 cells of different dose AD groups,the contents of SOD,T-AOC,the expression level of p-ERK were significantly increased,the proliferation inhibitory rate and apoptosis rate,the contents of IL-1β,TNF-α,IL-6,MDA,the expression level of p-p38 were significantly decreased(P<0.05).In MLE-12 cells of SB202190+AD group and PD98059+AD group,the contents of SOD,T-AOC were significantly decreased,the proliferation inhibitory rate and apoptosis rate,the contents of IL-1β,TNF-α,IL-6,MDA were significantly increased(P<0.05).The above results indicate that AD inhibits cell apoptosis,inflammatory response,and oxidative stress response b

关 键 词：甲型流感病毒 肺上皮细胞 穿心莲内酯 P38 细胞外调节蛋白激酶 

分 类 号：R373.13[医药卫生—病原生物学]