Cao Lu, Li Chun-Sun, Chang Yan, Liang Zhi-Xin, Chen Liang-An
Department of Respiratory Medicine, Chinese People's Liberation Army General Hospital, Beijing 100853, P.R. China.
Department of Respiratory Medicine, Chinese People's Liberation Army Second Artillery Force General Hospital, Beijing 100088, P.R. China.
Mol Med Rep. 2016 Apr;13(4):3700-8. doi: 10.3892/mmr.2016.4952. Epub 2016 Mar 1.
Acute lung injury (ALI)/ARDS is a critical clinical syndrome with high mortality, and the effective therapeutic methods for the treatment remain limited. Previous studies have indicated that liquid ventilation with perfluorocarbon (PFC) may be advantageous over conventional mechanical ventilation in the treatment of ALI/ARDS. Additionally, PFC inhibits the inflammatory response caused by ALI/ARDS. However, the anti-inflammatory mechanism remains to be completely elucidated. In the present study, the aim was to determine the anti‑inflammatory mechanism of PFC and the association with microRNA (miR). PFC was used to modulate LPS‑induced A549 cells, with the cells divided into four groups: Untreated control group; LPS group, treated with 10 µg/ml LPS; LPS+PFC group, treated with 10 µg/ml LPS and PFC; and PFC group, treated with PFC alone. The intercellular adhesion molecule‑1 (ICAM‑1) mRNA and protein expression levels of each group were detected by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blotting, respectively. A549 cells were transfected with miR‑17‑3p mimics, miR‑17‑3p inhibitors or negative controls to observe the alterations in the anti‑inflammatory effects of PFC. A dual luciferase reporter gene assay was used to determine whether ICAM‑1 is a target gene of miR‑17‑3p. PFC was observed to attenuate the mRNA and protein expression levels of ICAM‑1 in LPS‑induced A549 cells, with no significant effect on the untreated A549 cells. miR‑17‑3p was demonstrated to be regulated by PFC. Transfection with miR‑17‑3p mimics enhanced the anti‑inflammatory effects of PFC, whereas the miR‑17‑3p inhibitor weakened the anti‑inflammatory effects of PFC at early time points. To conclude, the current study indicates that ICAM‑1 was a target gene of miR‑17‑3p, and PFC has anti‑inflammatory effects. Additionally, the present study is the first report, to the best of our knowledge, that PFC is able to attenuate ICAM-1 expression in LPS-induced A549 cells by increasing miR-17-3p expression.
急性肺损伤（ALI）/急性呼吸窘迫综合征（ARDS）是一种死亡率很高的严重临床综合征，目前针对其有效的治疗方法仍然有限。先前的研究表明，全氟化碳（PFC）液体通气在治疗ALI/ARDS方面可能优于传统机械通气。此外，PFC可抑制ALI/ARDS引起的炎症反应。然而，其抗炎机制仍有待完全阐明。在本研究中，目的是确定PFC的抗炎机制及其与微小RNA（miR）的关系。使用PFC处理脂多糖（LPS）诱导的A549细胞，将细胞分为四组：未处理的对照组；LPS组，用10μg/ml LPS处理；LPS+PFC组，用10μg/ml LPS和PFC处理；PFC组，仅用PFC处理。分别通过逆转录-定量聚合酶链反应（RT-qPCR）和蛋白质印迹法检测每组细胞间黏附分子-1（ICAM-1）的mRNA和蛋白表达水平。用miR-17-3p模拟物、miR-17-3p抑制剂或阴性对照转染A549细胞，以观察PFC抗炎作用的变化。采用双荧光素酶报告基因测定法确定ICAM-1是否为miR-17-3p的靶基因。观察到PFC可降低LPS诱导的A549细胞中ICAM-1的mRNA和蛋白表达水平，而对未处理的A549细胞无显著影响。结果表明miR-17-3p受PFC调控。转染miR-17-3p模拟物可增强PFC的抗炎作用，而miR-17-3p抑制剂在早期时间点减弱了PFC的抗炎作用。总之，本研究表明ICAM-1是miR-17-3p的靶基因，且PFC具有抗炎作用。此外，据我们所知，本研究首次报道PFC能够通过增加miR-17-3p表达来减弱LPS诱导的A549细胞中ICAM-1的表达。
