a Department of Pathogen Biology, School of Medicine , Jinan University , Guangzhou , China.
b Department of Epidemiology, School of Medicine , Jinan University , Guangzhou , China.
Immunopharmacol Immunotoxicol. 2019 Apr;41(2):199-206. doi: 10.1080/08923973.2019.1569046. Epub 2019 Feb 6.
The industrial production and combustion of coal can produce silica nanoparticles (nano-SiO). It enters the human body mainly through the respiratory tract and exerts a toxic effect. However, whether nano-SiO can increase the IL-1β-induced inflammatory expression in A549 cells has not been tested. Therefore, the synergistic toxicity of nano-SiO and IL-1β to A549 was observed in our study. : We exposed A549 cells to nano-SiO (0, 100, 500, and 1000 μg/ml) for 12 and 24 h. The effect of nano-SiO on the viability of A549 cells was observed by the CCK-8 method. The A549 cells were exposed to nano-SiO (1 mg/mL) and cytokine IL-1β (10 ng/mL) for 4 h, and we detected the expression of IL-1β and IL-6 cytokines by real time quantitative polymerase chain (RT-qPCR) and enzyme linked immunosorbent assay (ELISA). The expression of β-Actin, I-κB, phospho-ERK (P-ERK), total-ERK (T-ERK), phospho-JNK (P-JNK), total-JNK (T-JNK), phospho-P38 (P-P38), and total-P38 (T-P38) in A549 cells was detected by the Western Blot method. The nano-SiO treatment resulted in a time-dependent decrease in the viability of A549 cells. The synergistic effect of nano-SiO and IL-1β was observed on the new production of IL-1β and IL-6 in A549 cells. The Western blot results showed that nano-SiO can increase the expression of IL-1β and IL-6 by promoting the phosphorylation of ERK and elevating the phosphorylation of I-κB by IL-1β. IL-1β and IL-6 were induced by nano-SiO, and the IL-1β treatment with 20 μM of I-κBα phosphorylation inhibitor (PD98059) and 20 μM of ERK inhibitor (BAY11-7082) for 1 h was significantly lower than that of the control group in A549 cells. These results indicated that nano-SiO had a toxic effect on A549 cells, and this effect could increase IL-1β on the A549 cell-induced inflammatory response. The results suggested that the release of IL-1β and IL-6 in A549 was enhanced by the synergistic IL-1β-induced phosphorylation of ERK and I-κB. This process is similar to a snowball, and it is possible that IL-1β is continuously produced and repeatedly superimposed in A549 cells to produce an inflammatory effect; then, a vicious circle occurs, and an inflammatory storm is accelerated.
工业生产和煤炭燃烧会产生二氧化硅纳米颗粒(nano-SiO)。它主要通过呼吸道进入人体,并产生毒性作用。然而,纳米 SiO 是否会增加 A549 细胞中 IL-1β 诱导的炎症表达尚未得到测试。因此,我们在研究中观察了纳米 SiO 和 IL-1β 对 A549 的协同毒性作用。
我们将 A549 细胞暴露于纳米 SiO(0、100、500 和 1000μg/ml)12 和 24 小时。通过 CCK-8 法观察纳米 SiO 对 A549 细胞活力的影响。将 A549 细胞暴露于纳米 SiO(1mg/ml)和细胞因子 IL-1β(10ng/ml)4 小时,通过实时定量聚合酶链(RT-qPCR)和酶联免疫吸附试验(ELISA)检测 IL-1β 和 IL-6 细胞因子的表达。通过 Western Blot 法检测 A549 细胞中 β-肌动蛋白、I-κB、磷酸化 ERK(P-ERK)、总 ERK(T-ERK)、磷酸化 JNK(P-JNK)、总 JNK(T-JNK)、磷酸化 P38(P-P38)和总 P38(T-P38)的表达。
纳米 SiO 处理导致 A549 细胞活力呈时间依赖性下降。在 A549 细胞中,观察到纳米 SiO 和 IL-1β 的协同作用导致新产生的 IL-1β 和 IL-6。Western blot 结果表明,纳米 SiO 可以通过促进 ERK 的磷酸化和升高 IL-1β 引起的 I-κB 的磷酸化来增加 IL-1β 和 IL-6 的表达。IL-1β 和 IL-6 被纳米 SiO 诱导,用 20μM I-κBα 磷酸化抑制剂(PD98059)和 20μM ERK 抑制剂(BAY11-7082)预处理 1 小时后,A549 细胞中 IL-1β 的表达明显低于对照组。
这些结果表明,纳米 SiO 对 A549 细胞具有毒性作用,这种作用可增强 IL-1β 对 A549 细胞诱导的炎症反应。结果表明,IL-1β 协同诱导的 ERK 和 I-κB 磷酸化增强了 A549 细胞中 IL-1β 和 IL-6 的释放。这一过程类似于滚雪球,在 A549 细胞中,IL-1β 可能会不断产生并反复叠加,从而产生炎症效应;然后,就会发生恶性循环,加速炎症风暴的发生。