Department of Physiology, College of Medicine, Kyung Hee University, Seoul, Republic of Korea.
Department of Pulmonary and Critical Care Medicine, Gangdong Kyung Hee University Hospital, Seoul, Republic of Korea.
J Biochem Mol Toxicol. 2021 Feb;35(2):e22635. doi: 10.1002/jbt.22635. Epub 2020 Sep 28.
Particulate matter (PM) of 10-μm-sized fine dust in the air penetrates the respiratory tract and contributes to the increasing incidence of various lung diseases, but its definite mechanism is not known. Recently, polydeoxyribonucleotide (PDRN) has been shown to have anti-inflammatory and regenerative effects in various tissues. However, the bronchial-related mechanism is not well-understood. Hence, this experiment is intended to demonstrate the beneficial effect of PDRN administration on PM10-induced injury in human bronchial-derived NCI-H358 cells. To confirm the protective effect of PDRN, PM10 was applied after PDRN pretreatment to confirm changes in NCI-H358 cells. Experiments were conducted to measure cell survival, cytotoxicity, inflammation, and apoptotic factor changes. WST-8 assay was used to confirm cell viability, and lactate dehydrogenase assay was used to obtain cytotoxicity. In addition, changes in inflammatory cytokines and apoptotic factors were confirmed by enzyme-linked immunosorbent assay and Western blot. Decreased cell viability and increased cytotoxicity, inflammatory cytokines, and apoptotic factors were observed after exposure to PM10. However, pretreatment with PDRN enhanced cell viability and reduced cytotoxicity. In addition, the expression of inflammatory cytokines such as tumor necrosis factor-α, interleukin-6 (IL-6), and IL-1β, and cell death factors such as Apaf-1, cyt c, caspase-3, caspase-9, Bid, and Bax/Bcl-2 ratio were decreased by PDRN administration in PM10-exposed NCI-H358 cells. PDRN, an A2AR agonist, affects cAMP activation and regulation of phosphorylation of PKA and CREB. In addition, treatment with A2AR antagonist 3,7-dimethyl-1-propargylxanthine significantly blocked PDRN's effect. These anti-cytotoxicity, anti-inflammation, and anti-apoptosis effects of PDRN can be attributed to the adenosine A2AR enhancing effect on PM10-exposed bronchial cells.
空气中 10μm 大小的细颗粒物（PM）会穿透呼吸道，导致各种肺部疾病的发病率上升，但具体机制尚不清楚。最近，聚脱氧核糖核苷酸（PDRN）已被证明在各种组织中具有抗炎和再生作用。然而，其在支气管方面的作用机制尚不清楚。因此，本实验旨在证明 PDRN 给药对 PM10 诱导的人支气管源性 NCI-H358 细胞损伤的有益作用。为了证实 PDRN 的保护作用，在给予 PDRN 预处理后应用 PM10，以确认 NCI-H358 细胞的变化。进行了实验以测量细胞存活、细胞毒性、炎症和凋亡因子变化。使用 WST-8 测定法确认细胞活力，使用乳酸脱氢酶测定法获得细胞毒性。此外，通过酶联免疫吸附试验和 Western blot 确认炎症细胞因子和凋亡因子的变化。暴露于 PM10 后，观察到细胞活力降低和细胞毒性增加、炎症细胞因子和凋亡因子增加。然而，PDRN 预处理增强了细胞活力并降低了细胞毒性。此外，PM10 暴露的 NCI-H358 细胞中，肿瘤坏死因子-α、白细胞介素-6（IL-6）和 IL-1β 等炎症细胞因子的表达以及 Apaf-1、细胞色素 c、caspase-3、caspase-9、Bid 和 Bax/Bcl-2 比值等细胞死亡因子的表达均降低。作为 A2AR 激动剂的 PDRN 影响 cAMP 的激活和 PKA 和 CREB 的磷酸化调节。此外，用 A2AR 拮抗剂 3,7-二甲基-1-丙炔基黄嘌呤处理可显著阻断 PDRN 的作用。PDRN 的这种抗细胞毒性、抗炎和抗凋亡作用可归因于对 PM10 暴露的支气管细胞的腺苷 A2AR 增强作用。
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