双氢青蒿素对甲型流感病毒H1N1诱导支气管上皮细胞中TNF-α和IL-6表达的影响及机制
[The Effects and Mechanisms of Dihydroartemisinin on Influenza A Virus H1N1 Induces TNF-α and IL-6 Expression in Bronchial Epithelial Cells].
作者信息
Ou Li, Qin Ke, Yang Zi-Xiao, Bie Ming-Jiang
机构信息
Department of Respiratory Medicine, Affiliated Chengdu 363 Hospital of Southwest Medical University, Chengdu 610041, China.
Department of Clinical Medicine, North Sichuan Medical College, Nanchong 637000, China.
出版信息
Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Mar;51(2):171-177. doi: 10.12182/20200360604.
OBJECTIVE
To investigate the effects of dihydroartemis (DHA) on influenza A virus (IAV) A/PR/8/34 (H1N1) induces the pro-inflammatory factor and protein of extracellular signal regulated kinase (ERK) signaling pathway expression in bronchial epithelial cells.
METHODS
The BEAS-2B cells were treated with different concentrations of DHA (i.e.,0， 12.5, 25，50 and 100 μmol/L) for 24 h and the effect of DHA on the viability of BEAS-2B cells were measure by CCK8 method. The BEAS-2B cells were absorbed with IAV for 1 h, and then were treated with different concentrations of DHA (i.e., 12.5, 25 and 50 μmol/L) for 24 h, meanwhile, the normal control group and IAV group were established. The mRNA and protein expression levels of tumor necrosis factor-α (TNF-α) and interleukin (IL-6) were measured by real time quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), the expression levels of phospho-ERK (p-ERK) proteins were tested by Western blot (WB). Then, an ERK agonist (20 ng/mL) was used to treat BEAS-2B cells (the groups were divided into normal control group, DHA group, DHA+IAV group, ERK agonist group and DHA+IAV+ERK agonist group) for 24 h, and to observe the effect of DHA on inhibiting IAV induce the TNF-α, IL-6 and p-ERK expression in the BEAS-2B cells.
RESULTS
The BEAS-2B cells viability was not significantly different from that of the normal control group after treatment with DHA (i.e., 12.5, 25, and 50 μmol/L). The expression levels of , -6 mRNA and TNF-α, IL-6, p-ERK protein in IAV group were significantly up-regulated compared with that in the normal control group ( <0.05), meanwhile, compared with the IAV group, the expression levels of , -6 mRNA and TNF-α, IL-6, p-ERK protein showed dose-dependent decrease in IAV+DHA group ( <0.05). However, ERK agonists attenuated the DHA inhibit IAV induced the proinflammatory factors TNF-α, IL-6 secretion and the p-ERK protein expression of ERK signaling pathway in BEAS-2B cells.
CONCLUSION
These data suggest that DHA can inhibit IAV induces the TNF-α and IL-6 expression in BEAS-2B cells through ERK signaling pathway.
目的
探讨双氢青蒿素（DHA）对甲型流感病毒（IAV）A/PR/8/34（H1N1）诱导支气管上皮细胞促炎因子及细胞外信号调节激酶（ERK）信号通路蛋白表达的影响。
方法
用不同浓度的DHA（即0、12.5、25、50和100μmol/L）处理BEAS-2B细胞24小时，采用CCK8法检测DHA对BEAS-2B细胞活力的影响。BEAS-2B细胞用IAV吸附1小时，然后用不同浓度的DHA（即12.5、25和50μmol/L）处理24小时，同时设立正常对照组和IAV组。采用实时定量PCR（RT-qPCR）和酶联免疫吸附测定（ELISA）检测肿瘤坏死因子-α（TNF-α）和白细胞介素（IL-6）的mRNA和蛋白表达水平，用蛋白质印迹法（WB）检测磷酸化ERK（p-ERK）蛋白的表达水平。然后，用ERK激动剂（20 ng/mL）处理BEAS-2B细胞（分为正常对照组、DHA组、DHA+IAV组、ERK激动剂组和DHA+IAV+ERK激动剂组）24小时，观察DHA对抑制IAV诱导BEAS-2B细胞中TNF-α、IL-6和p-ERK表达的作用。
结果
用DHA（即12.5、25和50μmol/L）处理后，BEAS-2B细胞活力与正常对照组相比无显著差异。与正常对照组相比，IAV组中TNF-α、IL-6 mRNA及TNF-α、IL-6、p-ERK蛋白表达水平显著上调（P<0.05），同时，与IAV组相比，IAV+DHA组中TNF-α、IL-6 mRNA及TNF-α、IL-6、p-ERK蛋白表达水平呈剂量依赖性降低（P<0.05）。然而，ERK激动剂减弱了DHA对IAV诱导BEAS-2B细胞中促炎因子TNF-α、IL-6分泌及ERK信号通路p-ERK蛋白表达的抑制作用。
结论
这些数据表明，DHA可通过ERK信号通路抑制IAV诱导BEAS-2B细胞中TNF-α和IL-6的表达。
Zotero 插件