School of Pharmacy, Fudan University, Shanghai, China; China State Institute of Pharmaceutical Industry, Shanghai, China; Pharmacology and Toxicology Department, Shanghai Institute for Food and Drug Control, Shanghai, China.
Pharmacology and Toxicology Department, Shanghai Institute for Food and Drug Control, Shanghai, China; NMPA Key Laboratory for Quality Analysis of Chemical Drug Preparations, Shanghai, China.
Pulm Pharmacol Ther. 2020 Aug;63:101939. doi: 10.1016/j.pupt.2020.101939. Epub 2020 Aug 27.
Inhalation of aerosolized drugs is a promising entry route for rapid and non-invasive therapeutics delivery to the lung. Curcumin exhibits potent anti-inflammatory properties, which are effective for use in lung diseases. The anti-inflammatory properties of curcumin have been widely studied in vitro with cells cultured in submerged conditions, however, the effectiveness using air-liquid interface (ALI) exposure is currently unknown.
The anti-inflammatory effect of curcumin under both ALI and submerged conditions was investigated in the present study. Lipopolysaccharide (LPS) stimulated A549 cells were exposed to curcumin under ALI (10-100 μM) using a dose-controlled air-liquid interface cell exposure (ALICE)-CLOUD system and submerged cell culture conditions (1-20 μM). The expression of pro-inflammatory cytokines (interleukin (IL)-6, IL-8), cell viability and cytotoxicity were studied for each exposure scenario. The cellular uptake behaviour of curcumin was studied with an equipotent cell-based dose (200 pmol/10 cell) at various time points up to 24 h.
The ALI delivery profile proved to be rapid, efficient and reproducible. For the doses studied, no significant effect on cell viability and cytotoxicity were observed. ALI exposure of curcumin was more effective in reducing pro-inflammatory cytokines expression in lung epithelial cells compared with submerged cell cultures. Furthermore, rapid cellular uptake and higher intracellular doses were achieved by ALI conditions.
The ALICE-CLOUD system combined with lung epithelial cells cultured under ALI conditions offers a reliable and relevant in vitro method for preclinical aerosolized drug screening. Curcumin might be a promising anti-inflammatory candidate drug for inhalation therapy of lung diseases.
吸入气溶胶药物是一种很有前途的途径，可以快速、非侵入性地将治疗药物递送到肺部。姜黄素具有很强的抗炎特性，可有效用于肺部疾病。姜黄素的抗炎特性已在体外进行了广泛研究，即在浸没条件下培养的细胞，但目前尚不清楚在气液界面 (ALI) 暴露下的效果。
本研究考察了姜黄素在 ALI 和浸没条件下的抗炎作用。采用剂量控制的气液界面细胞暴露（ALICE-CLOUD）系统和浸没细胞培养条件（1-20 μM），用脂多糖（LPS）刺激 A549 细胞，在 ALI 下（10-100 μM）暴露于姜黄素。研究了每种暴露情况的促炎细胞因子（白细胞介素 (IL)-6、IL-8）表达、细胞活力和细胞毒性。用等效细胞剂量（200 pmol/10 细胞）在 24 小时内的不同时间点研究了姜黄素的细胞摄取行为。
ALI 传递方式快速、高效且可重复。在所研究的剂量下，未观察到对细胞活力和细胞毒性有显著影响。与浸没细胞培养相比，ALI 暴露于姜黄素可更有效地降低肺上皮细胞中促炎细胞因子的表达。此外，通过 ALI 条件实现了快速的细胞摄取和更高的细胞内剂量。
结合在 ALI 条件下培养的肺上皮细胞的 ALICE-CLOUD 系统为临床前吸入药物筛选提供了一种可靠且相关的体外方法。姜黄素可能是肺部疾病吸入治疗有前途的抗炎候选药物。
