S100A9诱导单核细胞分泌血管内皮生长因子-A的初步机制研究

Study on the mechanism of S100A9-induced secretion of vascular endothelial growth factor-A by monocytes

目的探讨S100A9诱导单核细胞分泌血管内皮生长因子-A(VEGF-A)的分子机制。方法收集体检健康者外周血,利用免疫磁珠分选技术分离纯化出CD14+单核细胞,流式细胞术检测晚期糖基化终产物受体(RAGE)的表达,然后在体外加入S100A9刺激培养,或预先加入抗RAGE抗体或NK-κB信号抑制剂孵育1h后再进行刺激培养,酶联免疫吸附试验检测VEGF-A水平。结果S100A9受体RAGE高表达于单核细胞表面,S100A9刺激的单核细胞以剂量和时间依赖的方式分泌VEGF-A;加入抗RAGE抗体阻断后,单核细胞分泌VEGF-A的能力明显下降(P<0.01)。进一步利用NK-κB信号抑制剂可明显抑制S100A9诱导单核细胞分泌VEGF-A的作用(P<0.01)。结论 S100A9通过RAGE-NK-κB信号途径诱导单核细胞上调VEGF-A的分泌,从而有利于新生血管的生成。

Objective To explore the molecular mechanism of S100A9-induced secretion of vascular endothelial growth factor-A （VEGF-A）by monocytes.Methods Peripheral blood specimen were collected from healthy individuals undergoing physical exami-nation and the CD14 ＋ monocytes were purified by using immunomagnetic beads and the expression of the receptor for advanced gly-cation endproducts （RAGE）was detected by flow cytomertry.In vitro CD14 ＋ monocytes were stimulated by S100A9,and anti-RAGE antibody or NK-κB signal pathway inhibitor were pre-incubated for 1 hour and then stimulated by S100A9,the levels of VEGF-A were detected by using enzyme-linked immunosorbent assay.Results The high level of RAGE was expressed by isolated CD14 ＋ monocytes,after S100A9 stimulation,the secretion of VEGF-A by CD14 ＋ monocytes was significantly increased in a dose and time dependent manner.However,the inducing VEGF-A was significantly decreased（P 〈0.01 ）,while pre-treated with anti-RAGE antibody or NK-κB inhibitor （P 〈0.01）.Conclusion S100A9 inducing the secretion of VEGF-A by monocytes and is de-pended on RAGE-NK-κB signal pathway,suggesting that S100A9 might promote angiogenesis.