摘要
目的探讨米诺环素对急性肺损伤肺微血管内皮细胞炎症损伤的抑制作用。方法以脂多糖(LPS)10 mg·L^-1与肺微血管内皮细胞作用24 h,构建炎症损伤的细胞模型;米诺环素10和25μmol·L^-1共孵育24 h,用CCK-8法检测细胞存活率,ELISA检测培养液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-8和细胞间黏附分子1(ICAM-1)水平,流式细胞术分析细胞活性氧(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)和NF-κB表达。结果米诺环素可保护LPS诱导的肺微血管内皮细胞炎症损伤,LPS组细胞存活率为82.4%,米诺环素10和25μmol·L^-1组细胞存活率可达90.6%和96.9%(P<0.01)。米诺环素可降低LPS诱导的肺微血管内皮细胞炎症因子的表达,与LPS组相比,米诺环素组细胞TNF-α,IL-6,IL-8和ICAM-1的表达显著降低(P<0.01)。米诺环素还显著抑制LPS诱导的细胞氧化应激,LPS组细胞ROS相对水平为3.19,米诺环素10和25μmol·L^-1组为2.43和1.44(P<0.01)。Western印迹法检测结果显示,米诺环素可显著抑制LPS诱导炎症损伤的肺微血管内皮细胞PARP-1(P<0.01)和NF-κB表达(P<0.01)。结论米诺环素可有效抑制肺微血管内皮细胞的炎症损伤,其作用机制可能与抑制细胞PARP-1的表达、继而抑制NF-κB通路的活化、最终抑制炎症因子的表达有关。
OBJECTIVE To investigate the protective effect of minocycline against inflammatory injury to pulmonary microvascular endothelial cells in acute lung injury.METHODS An in vitro model of inflammatory injury in pulmonary microvascular endothelial cells was induced by lipopolysaccharide(LPS)10 mg·L^-1 for 24 h.The anti-inflammatory group was incubated with minocycline 10 and 25μmol·L^-1 for 24 h.Cell viability was assayed by CCK-8,the levels of tumor necrosis factor-α(TNF-α),interleukin 6(IL-6),IL-8 and intercellular cell adhesion molecule-1(ICAM-1)were detected by ELISA,the level of reactive oxygen species(ROS)was analyzed by flow cytometry,the expression of poly(ADP-ribose)polymerase-1(PARP-1)and NF-κB signaling pathway in cells were detected by Western blotting.RESULTS Minocycline could effectively protect the pulmonary microvascular endothelial cells from inflammatory damage induced by LPS.The cell viability of the minocycline 10 and 25μmol·L^-1 groups was 90.6%and 96.9%(P<0.01),compared with 82.4%in the LPS group.Minocycline could reduce the levels of inflammatory cytokines in pulmonary microvascular endothelial cel s induced by LPS.The expressions of TNF-α,IL-6,IL-8 and ICAM-1 in the minocycline group were significantly lower than those in the LPS group(P<0.01).Minocycline could also significantly inhibit the oxidative stress in pulmonary microvascular endothelial cel s induced by LPS.The ROS relative level of LPS group was 3.19,compared with 2.43 and 1.44 in the minocycline groups(P<0.01).Western blotting results showed that minocycline significantly inhibited the expression of PARP-1(P<0.01)and activation of NF-κB pathway(P<0.01)in pulmonary microvascular endothelial cells induced by LPS.CONCLUSION Minocycline can effectively inhibit the inflammatory damage to pulmonary microvascular endothelial cells.The mechanism may be related to the inhibited expression of PARP-1,thus inhibiting the activation of NF-κB pathway and the expression of inflammatory cytokines.
作者
罗超
吴伟斌
林欣雨
祝春燕
LUO Chao;WU Wei-bin;LIN Xin-yu;ZHU Chun-yan(Yuanpei College,Shaoxing University,Shaoxing 312000,China;Zhaoqing Medical College,Zhaoqing 526020,China;Affiliated Hospital,Shaoxing University,Shaoxing 312000,China)
出处
《中国药理学与毒理学杂志》
CAS
北大核心
2020年第9期656-662,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)。
