Kucukgul Altug, Erdogan Suat
a Department of Biochemistry , Veterinary Faculty, Mustafa Kemal University , Hatay , Turkey.
b Department of Medical Biology , Faculty of Medicine, Trakya University , Edirne , Turkey.
Exp Lung Res. 2017 Feb;43(1):1-7. doi: 10.1080/01902148.2016.1267823. Epub 2017 Jan 12.
The current study aimed to investigate in vitro effects of oleic acid on lipopolysaccharide (LPS)-induced acute lung injury in the human lung epithelial cells (A549).
The cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tests. Selected gene expression levels were analyzed by Real-Time Quantitative-Polymerase Chain Reaction (RT-qPCR).
24 hours of LPS (100 ng/mL) exposure decreased the cells' viability by 44.6% compared to untreated control. Low concentration (2.5 nM) of oleic acid slightly suppressed the cell survival by 9.1% analyzed 24 hours after incubation. However, oleic acid pretreatment before LPS exposure significantly increased cell survival loss to 63.9%. LPS exposure decreased the expressions of catalase (CAT) and glutathione peroxidase (GPx) mRNA levels by 2.8 and 2.5 fold, respectively. Moreover, pretreatment of the cells with oleic acid strengthened LPS-decreased expressions of CAT and GPx genes by 3.5 and 6.7 fold, respectively. The mRNA expressions of superoxide dismutase (SOD), induced nitric oxide synthase (iNOS), interleukin-1β, IL-12, COX-2, caspase-3 and caspase-8 were increased by 2.4, 2.2, 2.2, 2.3, 3.0, 2.6, and 2.5 fold, respectively, by LPS, and oleic acid pretreatment significantly potentiated the effect of LPS.
Oleic acid worsens LPS-induced cell death by potentiating oxidative stress and inflammation in A549 lung epithelial cells.
本研究旨在探讨油酸对脂多糖（LPS）诱导的人肺上皮细胞（A549）急性肺损伤的体外影响。
通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）试验评估细胞活力。通过实时定量聚合酶链反应（RT-qPCR）分析选定基因的表达水平。
与未处理的对照组相比，暴露于100 ng/mL LPS 24小时后，细胞活力降低了44.6%。孵育24小时后分析发现，低浓度（2.5 nM）的油酸轻微抑制细胞存活，抑制率为9.1%。然而，在LPS暴露前用油酸预处理显著增加了细胞存活损失至63.9%。LPS暴露使过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GPx）的mRNA水平分别降低了2.8倍和2.5倍。此外，用油酸预处理细胞分别使LPS降低的CAT和GPx基因表达增强了3.5倍和6.7倍。超氧化物歧化酶（SOD）、诱导型一氧化氮合酶（iNOS）、白细胞介素-1β、IL-12、环氧化酶-2、半胱天冬酶-3和半胱天冬酶-8的mRNA表达分别因LPS而增加了2.4、2.2、2.2、2.3、3.0、2.6和2.5倍，油酸预处理显著增强了LPS的作用。
油酸通过增强A549肺上皮细胞中的氧化应激和炎症反应，加重LPS诱导的细胞死亡。
