摘要
目的探讨奥拉帕尼对脂多糖(LPS)诱导的肺泡上皮细胞炎症损伤的保护作用。方法体外培养的肺泡上皮细胞(A549)同时加入LPS 10mg·L^-1+奥拉帕尼10和25 μmol·L^-1孵育24 h。酶联免疫吸附试验(ELISA)测定白细胞介素6(IL-6),IL-8和IL-10释放;实时PCR技术检测肿瘤坏死因子α(TNF-α)、IL-1β、IL-6、IL-8和细胞间黏附分子1(ICAM-1)mRNA 的表达水平;流式细胞术检测细胞活性氧(ROS)水平;Western印迹法检测细胞聚腺苷二磷酸核糖聚合酶1(PARP-1)蛋白表达水平和NF-κB通路相关蛋白磷酸化水平。结果与LPS 10 mg·L^-1损伤组相比,奥拉帕尼10和25 μmol·L^-1显著降低LPS诱导的A549细胞IL-6,IL-8的释放(P<0.01)和ROS水平(P<0.01),升高IL-10水平(P<0.01),抑制了LPS诱导的TNF-α,IL-1β,IL-6,IL-8和ICAM-1 mRNA表达水平(P<0.01),抑制LPS诱导的PARP-1蛋白表达和NF-κB通路磷酸化激活(P<0.01)。结论奥拉帕尼可有效缓解LPS诱导的肺泡上皮细胞炎症损伤和氧化应激,其作用机制可能为奥拉帕尼通过下调PARP-1的表达,继而影响NF-κB通路的活化,最终抑制了相关炎症因子的表达和释放。
OBJECTIVE To investigate the protective effect of olaparib on the inflammatory damage to alveolar epithelial cells induced by lipopolysaccharide (LPS). METHODS The alveolar epithelial cells (A549) were cultured in vitro and incubated with LPS 10 mg·L^-1 and olaparib 10 and 25 μmol·L^-1 for 24 h. The levels of cytokines interleukin 6 (IL-6), IL-8, and IL-10 were detected by enzyme linked immunosorbent assay (ELISA), the mRNA expression levels of TNF-α, IL-1β, IL-6, IL-8, and ICAM-1 were analyzed by real-time PCR, the level of ROS was analyzed by flow cytometry, and the expression of poly (ADP-ribose) polymerase-1 (PARP-1) and phosphorylation of proteins involved in NF-κB signaling pathway in cells were detected by Western blotting. RESULTS Compared with LPS 10 mg·L^-1 injury group, olaparib 10 and 25 μmol·L^-1 could significantly reduce the release of IL-6, IL-8 and ROS levels in A549 cells induced by LPS (P<0.01), and increase the release of IL-10 (P<0.01). Olaparib 10 and 25 μmol·L^-1 could also inhibit the mRNA expressions of TNF-α, IL-1β, IL-6, IL-8, and ICAM-1 (P<0.01), and inhibit the expression of PARP-1 and phosphorylation proteins involved in NF-κB signaling pathway induced by LPS (P<0.01). CONCLUSION Olaparib has some protective effect on inflammatory damage and oxidative stress in alveolar epithelial cells induced by LPS, and the mechanism may be that it inhibits the expression and release of cytokines by down-regulating the expression of PARP-1 and subsequently affecting the activation of the NF-κB pathway.
作者
罗超
吴伟斌
周瑾
魏熵熵
厉美洋
LUO Chao;WU Wei-bin;ZHOU Jin;WEI Shang-shang;LI Mei-yang(Shaoxing University Yuanpei College, Shaoxing 312000, China;Zhaoqing Medical College, Zhaoqing 526020, China)
出处
《中国药理学与毒理学杂志》
CAS
北大核心
2019年第3期193-199,共7页
Chinese Journal of Pharmacology and Toxicology
基金
浙江省自然科学基金(LQ16H310002)
绍兴市科技计划项目(2018C30008)~~
