【摘要】 目的研究磷酸二酯酶(PDE4)抑制剂对细菌脂多糖(LPS)诱导的急性肺损伤(ALI)炎症反应的影响。方法离体细胞实验:原代培养肺微血管内皮细胞(PMVECs)并传代,用PDE4抑制剂西洛司特(cilomilast)和地西泮(diazepam)进行预处理后,加入LPS刺激细胞,制备ALI模型,将实验分为空白对照组(Ctrl),LPS+cilomilast(LPS+Cilo)组,LPS+diazepam(LPS+Diaz)组和LPS 2μg/mL组(LPS),分别测定核因子(NF-κB)转录活性、PDE4活性、PDE4B蛋白表达以及炎性因子MCP-1、IL-8、ICAM-1、VCAM-1的表达。在体细胞实验:4周龄雄性C57BL/6小鼠,体重17～25 g,分别用二甲基亚砜(DMSO)、cilomilast和diazepam预处理,30天后,使用LPS制备ALI模型,将实验分为DMSO+气道内滴注等容积生理盐水(Ctrl组),DMSO+气道内滴注LPS(LPS组),cilomilast+气道内滴注LPS(Cilo组),diazepam+气道内滴注LPS(Diaz组),分别测定PDE4B、ICAM-1、VCAM-1蛋白表达及IL-8、MCP-1的水平。结果 LPS刺激引起NF-κB、PDE4活性增加,PDE4B蛋白、MCP-1、IL-8、ICAM-1、VCAM-1表达增强;cilomilast和diazepam剂量依赖性减弱LPS引起的上述各项指标的增加。结论 PDE4抑制剂能够减轻急性肺损伤的炎症反应,可能与抑制NF-κB信号转导途径有关。更多还原

【Abstract】 Objective To study the effect of phosphodiesterase(PDE4) inhibitor on bacterial lipopolysaccharide(LPS) which induced inflammatory response to acute lung injury(ALI). Methods In vitro cells: Primary culture of pulmonary microvascular endothelial cells(PMVECs) and then subcultured, after pretreatmented with PDE4 inhibitor cilomilast and diazepam, added LPS to stimulate cells to prepare ALI model. The experiment was divided into blank control group(Ctrl), LPS+cilomilast(LPS+Cilo) group, LPS+diazepam(LPS+Diaz) group and LPS 2 μg/mL group(LPS). Nuclear factor-κB transcriptional activity, PDE4 activity, PDE4 B protein expression and the expressions of inflammatory factors including MCP-1, Il-8, ICAM-1 and VCAM-1 were detected. In somatic cells: 4 weeks old male C57 BL/6 mices, weight 17～25 g, ALI models were prepared by LPS after 30 days of pretreatment with DMSO, cilomilast and diazepam, respectively. The experiment was divided into DMSO+intratracheal saline(Ctrl), DMSO+intratracheal LPS(LPS), cilomilast + intratracheal LPS(Cilo), diazepam+intratracheal LPS(Diaz). Protein expression of PDE4 B, MCP-1, Il-8, ICAM-1 and VCAM-1 were detected respectively. Results LPS stimulation increased the activity of NF-κB and PDE4, and increased the expression of PDE4 B protein, MCP-1, Il-8, ICAM-1 and VCAM-1. Cilomilast and diazepam decreased the above indicators. Conclusion PDE4 inhibitors can alleviate the inflammatory reaction of acute lung injury, which may be related to the inhibition of NF-κB signal transduction pathway.更多还原