白细胞介素-1β促人视网膜色素上皮细胞分泌血管内皮生长因子和碱性成纤维细胞生长因子的研究  

Interleukin-1β stimulates the secretion of vascular endothelial growth factor and basic fibroblast growth factor in human retinal pigment epithelial cells

作　　者：李莹[1] 张潇[1] 赵潺[1] 龙琴[1] 

机构地区：[1]中国医学科学院北京协和医院眼科,100730

出　　处：《中华眼底病杂志》2009年第5期376-379,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（30600692）

摘　　要：目的 观察炎前因子白细胞介素-1β（IL-1β）对培养的人视网膜色素上皮（RPE）细胞分泌血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）的影响。方法采用酶联免疫吸附测定法（ELISA）测定IL—G浓度为0、1、10、20ng／ml和时间为24、36、48h作用下RPE细胞的VEGF和bFGF分泌量变化；采用半定量逆转录聚合酶链式反应（RTPCR）测定10ng／ml IL-1β作用48h后RPE细胞的VEGF和bFGFmRNA变化；噻唑蓝（MTT）比色法测定IL-1β刺激后RPE细胞的增生状态。结果相同刺激时间即36h作用下，终浓度为1、10ng／ml的IL-1β促进RPE细胞VEGF的分泌（q=32．79，42．56；P〈0．01）；10ng／ml的Il-1β促进bFGF的分泌（q=7．514，P＜0．01）；与浓度为10ng／ml的IL-1β刺激相比，增加IL-1β刺激浓度至20ng／ml使RPE细胞VEGF和bFGF分泌量下降（q=9．35，6．92；P＜0．01）。相同浓度即10ng／ml的IL-1β作用下IL-1β作用时间与RPE细胞VEGF和bFGF分泌量具有正性时间-效应关系；浓度为10ng／ml的IL-1β作用48h可促进RPE细胞的VEGF和bFGFmRNA水平；MTT结果证明IL-1β未明显影响RPE细胞的增生状态（F=0．317，P〉0．05）。结论炎前因子IL-1β可影响人RPE细胞VEGF和bFGF的分泌量，在一定的作用浓度和作用时间范围内具有显著的促进效应。Objective To investigate the effect of interleukin （IL-1β） on vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） secretion in cultured human retinal pigment epithelial （RPE） cells. Methods Enzyme linked immunosorbent assay （ELISA） was used to detect the production of VEGF and bFGF by RPECs. The cells were cultured in the presence of IL-1β at different concentrations,which were 0, 1,10, and 20 ng/ml, respectively. VEGF and bFGF levels were measured at various time points. Semiquantitative reverse transcription-polymerase chain reaction （RT-PCR） was conducted todetect the expression of VEGF and bFGF mRNA in RPE cells with and without IL-1β stimulation. The proliferation of cells in all groups was measured by the MTT assay and expressed as light absorption values. Results IL-1β significantly stimulated the secretion of VEGF at the concentration of 1 ng/ml and 10 ng/ml （P〈0.01） ,bFGF was significantly stimulated when the concentration of IL-1β reached 10 ng/ml （P〈0. 01） ;at the concentration of 10 ng/ml, IL-1β increased the production of VEGF and bFGF in a time dependent manner （P%0. 01）; VEGF and bFGF mRNA levels were significantly increased by stimulation of 10 ng/ml IL-1β for 48 hours. There was no significant difference between all groups in light absorption values （P〉0. 05）. Conclusion Pro-inflammatory cytokine IL-1β increased the secretion of VEGF and bFGF by human RPE cells to some extent.

关 键 词：色素上皮 眼/生理学 自细胞介素1β/投药和剂量 血管内皮生长因子类 成纤维细 胞生长因子2 

分 类 号：R774.1[医药卫生—眼科]