高迁移率族蛋白B1对人原代支气管上皮细胞血管内皮生长因子表达和释放的影响

发布时间：2018-02-11 05:34

本文关键词： 支气管哮喘高迁移率族蛋白B 原代支气管上皮细胞血管内皮生长因子　出处：《广东医学》2014年22期 　论文类型：期刊论文

【摘要】：目的探讨高迁移率族蛋白B1(HMGB1)对人原代支气管上皮细胞(PBEC)血管内皮生长因子(VEGF)表达和释放的影响及可能调控机制。方法 (1)按浓度依赖性分组;给予HMGB1 0、100、500、2 000μg/L刺激细胞;(2)时间依赖性分组:设置空白对照,HMGB1 2 000μg/L刺激PBEC时间为6、12、24 h。四唑盐法检测上述方法干预PBEC后细胞活力;实时荧光定量PCR检测PBEC VEGF mRNA表达水平变化,双抗体夹心酶联免疫吸附实验法检测细胞培养上清液中VEGF的浓度。拮抗高级糖基化终产物受体(RAGE)观察其对HMGB1诱导表达和释放VEGF的影响。结果 HMGB1在不影响细胞活力的情况下,(1)HMGB1与PBEC VEGF的表达和分泌水平存在剂量-效应关系,HMGB1(500、2 000μg/L)浓度组与对照组比较差异有统计学意义(P0.05)。(2)随HMGB1 2 000μg/L刺激时间延长,VEGF的表达和分泌逐渐升高,呈时间依赖性,且12、24 h组与对照组比较差异有统计学意义(P0.05)。(3)HMGB1 2 000μg/L刺激PBEC 24 h显著增加VEGF表达和释放,拮抗RAGE后显著抑制了HMGB1诱导的VEGF表达和释放。结论 HMGB1通过受体RAGE途径的增加PBEC VEGF的表达和释放,从而参与慢性气道炎症的发生和发展,其相关分子机制仍需要进一步探讨。
[Abstract]:Objective to investigate the effect of high mobility group protein B1 (HMGB1) on the expression and release of vascular endothelial growth factor (VEGF) in human primary bronchial epithelial cells (PBECs) and its possible regulatory mechanisms. HMGB1 0 / 100g / L #number0# 渭 g / L stimulated cells were divided into two groups in a time-dependent manner: HMGB12 000 渭 g / L PBEC stimulation time was 6 ~ 12 ~ 24 h. The cell viability was detected by tetrazolium tetrazolium method, and the expression of PBEC VEGF mRNA was detected by real-time fluorescence quantitative PCR, and the expression of PBEC VEGF mRNA was measured by real-time fluorescence quantitative PCR. Double antibody sandwich enzyme-linked immunosorbent assay (Elisa) was used to detect the concentration of VEGF in cell culture supernatant. Antagonistic advanced glycosylation end product receptor (ragger) was used to observe the effect on HMGB1 induced expression and release of VEGF. Results HMGB1 had no effect on cell viability. There was a dose-effect relationship between HMGB1 and the expression and secretion level of PBEC VEGF. The expression and secretion of HMGB1 in HMGB1 / 500000 渭 g / L group were significantly higher than those in the control group (P0.05 / L). The expression and secretion of HMGB1 increased gradually with the prolongation of the stimulation time of HMGB1 2 000 渭 g / L. In a time-dependent manner, there was a significant difference between the 12h group and the control group. The expression and release of VEGF were significantly increased by stimulation of PBEC at #number0# 渭 g / L for 24 h. After antagonizing RAGE, VEGF expression and release induced by HMGB1 were significantly inhibited. Conclusion the expression and release of PBEC VEGF through the receptor RAGE pathway are increased by HMGB1, which is involved in the occurrence and development of chronic airway inflammation, and its molecular mechanism needs to be further explored.
【作者单位】：广西医科大学第一附属医院呼吸内科;
【基金】：广西研究生教育创新计划项目(编号:YCBZ2013014)
【分类号】：R562.25

【相似文献】