Shanghai Key laboratory of Atmospheric Particle Pollution Prevention (LAP3), Department of Environmental Science & Engineering, Fudan University, 220 Handan Road, Shanghai 200433, PR China.
Shanghai Key laboratory of Atmospheric Particle Pollution Prevention (LAP3), Department of Environmental Science & Engineering, Fudan University, 220 Handan Road, Shanghai 200433, PR China; Engineering Research Center of Optical Instrument and System, Ministry of Education, Shanghai Key Lab of Modern Optical System, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, PR China.
Sci Total Environ. 2019 Aug 10;677:108-119. doi: 10.1016/j.scitotenv.2019.04.203. Epub 2019 Apr 16.
Ambient fine particle is a crucial indicator of air pollution brought into the air by sundry natural and public events. However, a comprehensive understanding of the PM-induced cytotoxicity especially the contribution of bioaerosol part is still undiscovered. Herein, an ALI microfluidics system integrated multi-omics (iTRAQ & RNA-seq) was successfully utilized to recognize the molecular mechanisms induced by microorganisms carried bioaerosol in human lung epithelial cells. The cells viability was above 98% within 21 days on this system. Moreover, the results showed that eight microorganisms-related pathways (e.g., Salmonella, amoebiasis, HTLV-1) were activated after exposure to PM for 24 h, which played a certain proportion in contributing to inflammation reaction. In addition, multi-omics demonstrated that three inflammation-related signal transduction cascades including MAPK signaling pathway, TNF signaling pathway, and TGF signaling pathway were triggered by fine particles, ultimately leading to apoptosis-related process disorder by associated cytokines like TNF, IL6, and TGF-β. Furthermore, flow cytometry analysis showed that the cell apoptosis rate increased from 3.8% to 66.7% between the cells exposed to PM (10 μg/cm) for 24 h and untreated control cells, which indicated that the fine particles had the ability to activate apoptosis-related signal cascades and result in apoptosis. ELISA assay and western blot indicated that HO-1, JNK, IL6, TNF, NF-κB, and FGF14 were significantly increased after exposure to PM while Casp3 and FGFR were decreased, which were consistent with the multi-omics. Moreover, PM components (OC, EC, 16PAHs, As, Cu, Mn, Cl, and NO) were significantly correlated to the inflammation related proteins and cytokines, which played a vital role in the inflammation and apoptosis related signaling pathways. These findings pointed to strong links among microorganisms infection, inflammation, and apoptosis in cell response to PM carried microorganisms. It also provided a new approach for understanding PM-induced cytotoxicity and health risks.
环境细颗粒物是各种自然和人为事件带入空气中的空气污染的一个关键指标。然而，人们对 PM 诱导的细胞毒性，特别是生物气溶胶部分的贡献，仍缺乏全面的认识。在此，我们成功地利用了一种整合了多组学（iTRAQ 和 RNA-seq）的 ALI 微流控系统，来识别微生物携带的生物气溶胶在人肺上皮细胞中引起的分子机制。在这个系统上，细胞在 21 天内的存活率超过 98%。此外，结果表明，暴露于 PM 24 小时后，有 8 种与微生物相关的途径（如沙门氏菌、阿米巴病、HTLV-1）被激活，这些途径在炎症反应中起一定作用。此外，多组学研究表明，MAPK 信号通路、TNF 信号通路和 TGF 信号通路这三个与炎症相关的信号转导级联反应被细颗粒物触发，最终导致与 TNF、IL6 和 TGF-β等相关细胞因子有关的细胞凋亡相关过程紊乱。此外，流式细胞术分析表明，暴露于 PM（10μg/cm）24 小时的细胞凋亡率从 3.8%增加到 66.7%，而未经处理的对照细胞凋亡率没有变化，这表明细颗粒物具有激活细胞凋亡相关信号级联反应和导致细胞凋亡的能力。ELISA 检测和 Western blot 表明，暴露于 PM 后，HO-1、JNK、IL6、TNF、NF-κB 和 FGF14 显著增加，而 Casp3 和 FGFR 减少，这与多组学结果一致。此外，PM 成分（OC、EC、16PAHs、As、Cu、Mn、Cl 和 NO）与炎症相关蛋白和细胞因子显著相关，在炎症和细胞凋亡相关信号通路中发挥着重要作用。这些发现表明，在细胞对携带微生物的 PM 做出反应时，微生物感染、炎症和细胞凋亡之间存在着紧密联系。它还为理解 PM 诱导的细胞毒性和健康风险提供了一种新的方法。
