丹皮酚抑制PM2.5诱导的人支气管上皮细胞VEGF表达的研究

摘要

目的 观察丹皮酚对细颗粒物(particulate matter 2.5,PM2.5)诱导的人支气管上皮细胞(BE-AS-2B)表达血管内皮生长因子(vascular endothelia growth factor,VEGF)的影响.方法 选取不同浓度PM2.5(25μg/mL,50μg/mL,100μg/mL)刺激BEAS-2B细胞,分别收集刺激6 h、12 h、24 h后的细胞培养上清液和刺激24 h后的细胞,使用酶联免疫吸附试验(ELISA)和Western Blot方法检测上清液中及细胞内VEGF的表达水平;利用不同浓度丹皮酚(15μmol/L,30μmol/L)进行干预24 h,收集细胞培养上清液,采用ELISA方法检测上清液中VEGF的表达水平.结果 随着PM2.5浓度的升高,细胞培养上清及支气管上皮细胞中VEGF蛋白表达水平呈剂量依赖性增高;随着PM2.5作用时间的延长,细胞培养上清中VEGF的表达水平呈时间依赖性增高;丹皮酚干预可抑制PM2.5诱导的VEGF表达,且抑制效应呈剂量依赖性.结论 PM2.5可诱导BEAS-2B表达VEGF,该作用可以被丹皮酚抑制.%Objective To explore the effect of peaonol on the expression of vascular endothelia growth fac-tor (VEGF) in human epithelia cell (BEAS-2B) stimulated with particulate matter 2. 5 (PM2. 5). Methods BE-AS-2B cells were stimulated with PM2. 5 (25 μg/mL, 50 μg/mL, 100 μg/mL), the cell culture supernatants were collected after 6 h, 12 h and 24 h and cells were obtained after 24 h. VEGF in supernatants and cells were detected with enzyme-linked immunosorbent assay ( ELISA) and Western Blot respectively. BEAS-2B cells stimulated with PM2. 5 (25 μg/mL, 50 μg/mL, 100μg/mL) were treated with peaonol (15μmol/L, 30μmol/L) for 24 h in vitro. After treatment, the cell culture supernatants were collected and VEGF in the supernatants were estimated by ELISA. Results PM2. 5 enhanced the protein expression of VEGF in supernatants and BEAS-2B cells in a dose dependent way and increased its expression in supernatants in a time dependent way. Peaonal could inhibit PM2. 5-induced VEGF expression in a dose dependent way. Conclusion PM2. 5 could up-regulate the expression of VEGF in BEAS-2B, which could be suppressed by peaonol.