INSERM, U968, Paris, France.
PLoS One. 2013 Aug 19;8(8):e72459. doi: 10.1371/journal.pone.0072459. eCollection 2013.
To characterize the effects of benzalkonium chloride (BAK) in THP-1 differentiated cells in vitro.
Macrophages were obtained after differentiation of THP-1 cells, a human monocytic leukemia cell line. Macrophages were exposed for 24 h to 33 nM (10(-5)%) benzalkonium chloride (BAK), 10 nM dinitrochlorobenzene (DNCB), 100 ng/mL lipopolysaccharide (LPS), 5 ng/mL tumor necrosis factor alpha (TNF-α) or phosphate buffered saline (PBS) as controls. The expression of CD11b, CD11c, CD33 and CD54 was evaluated using immunohistochemistry and flow cytometry (FCM). Phagocytosis function was analyzed using carboxylate-modified fluorescent microspheres and quantified by FCM. Migration was evaluated in cocultures with conjunctival epithelial cells. Cytokine production was detected and quantified in culture supernatants using a human cytokine array.
Stimulation of THP-1-derived macrophages with a low concentration of BAK increased CD11b and CD11c expression and decreased CD33. Macrophages exposed to BAK, LPS and TNF-α had increased phagocytosis. In contrast to LPS, BAK and TNF-α increased macrophage migration. Cytokines in supernatants of macrophages exposed to BAK revealed an increased release of CCL1, CCL4/MIP-1β, TNF-α, soluble CD54/ICAM-1 and IL-1β.
In vitro, BAK has a direct stimulating effect on macrophages, increasing phagocytosis, cytokine release, migration and expression of CD11b and CD11c. Long-term exposure to low concentrations of BAK should be considered as a stimulating factor responsible for inflammation through macrophage activation.
研究苯扎氯铵（BAK）对体外 THP-1 分化细胞的影响。
用 THP-1 细胞系（一种人单核白血病细胞）分化得到巨噬细胞，将巨噬细胞暴露于 33 nM（10(-5)%)苯扎氯铵（BAK）、10 nM 二硝基氯苯（DNCB）、100 ng/ml 脂多糖（LPS）、5 ng/ml 肿瘤坏死因子-α（TNF-α）或磷酸盐缓冲盐水（PBS）中 24 h，作为对照。采用免疫组织化学和流式细胞术（FCM）检测 CD11b、CD11c、CD33 和 CD54 的表达。用羧基化荧光微球分析吞噬功能，并通过 FCM 进行定量。在与结膜上皮细胞的共培养中评估迁移。采用人细胞因子芯片检测培养上清液中细胞因子的产生和定量。
低浓度 BAK 刺激 THP-1 衍生的巨噬细胞增加 CD11b 和 CD11c 的表达，降低 CD33。暴露于 BAK、LPS 和 TNF-α的巨噬细胞吞噬作用增强。与 LPS 不同，BAK 和 TNF-α增加了巨噬细胞迁移。暴露于 BAK 的巨噬细胞上清液中的细胞因子显示 CCL1、CCL4/MIP-1β、TNF-α、可溶性 CD54/ICAM-1 和 IL-1β 的释放增加。
体外，BAK 对巨噬细胞有直接的刺激作用，增加吞噬作用、细胞因子释放、迁移和 CD11b 和 CD11c 的表达。长期暴露于低浓度的 BAK 应被视为通过巨噬细胞激活引起炎症的刺激因素。
