β2-糖蛋白Ⅰ对VEGF诱导的巨噬细胞迁移的影响  

Effects of β2-glycoprotein Ⅰ on the migration of macrophage induced by VEGF

作　　者：宋振强[1] 王靖宇[1] 周赛君[1] 于珮[1] Song Zhenqiang;Wang Jingyu;Zhou Saijun;Yu Pei(Tianjin Medical University Chu Hsien-ⅠMemorial Hospital,NHC Key Laboratory of Hormones and Development(Tianjin Medical University),Tianjin Key Laboratory of Metabolic Diseases,Tianjin 300070,China)

机构地区：[1]天津医科大学朱宪彝纪念医院,国家卫生健康委员会激素与发育重点实验室,天津市代谢性疾病重点实验室,300070

出　　处：《国际内分泌代谢杂志》2020年第5期304-309,F0003,共7页International Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金(30971393)

摘　　要：目的观察血管内皮生长因子(VEGF)对小鼠单核/巨噬细胞系RAW 264.7的趋化作用,以及β2-糖蛋白Ⅰ(论-GP)对VEGF诱导巨噬细胞迁移的影响,探讨其可能机制。方法体外培养RAW 264.7小鼠单核/巨噬细胞系,实验分为5组:空白对照组、VEGF组、VEGF+β2-GP组、VEGF+SB203580(p38抑制剂)组、β2-GP组。Transwell实验观察β2-GP对VEGF诱导巨噬细胞迁移的影响,ELISA法检测P2-GP对VEGF诱导巨噬细胞分泌趋化因子单核细胞趋化蛋白(MCP)-1、白细胞介素(IL)-8以及巨噬细胞炎性蛋白-β(MIP-1β)的影响。Western印迹检测β2-GP对VEGF干预下巨噬细胞VEGF受体1(VEGFR1)和p38丝裂原活化蛋白激酶(MAPK)信号通路的影响。结果(1)与空白对照组相比,VEGF组巨噬细胞迁移的数量增加(t=22.089,P<0.05),与VEGF组相比,VEGF+β2-GP组和VEGF+SB203580组巨噬细胞迁移明显降低(t=22.687,63.625,P均<0.05)。(2)与空白对照组相比,VEGF组巨噬细胞趋化因子MCP-1、IL-8及MIP-1β的分泌增加(t=3.339,11.140,2.254,P均<0.05),与VEGF组相比,VEGF+β2-GP组和VEGF+SB203580组抑制VEGF诱导的3种趋化因子的分泌减少(t=3.148、2.402、2.798、6.754,4.459、5.528,P均<0.05)。(3)与空白对照组相比,VEGF组巨噬细胞VEGFR1的表达及p38MAPK通路的磷酸化程度增加(t=5.161,P<0.05);与VEGF组相比,VEGF+β2-GP组巨噬细胞VEGFR1表达及p38MAPK通路的磷酸化程度降低(t=4.965,P<0.05)c结论VEGF通过促进巨噬细胞VEGFR1表达,活化p38MAPK,促进巨噬细胞的迁移以及趋化因子MCP-1、IL-8,MIP-1β的分泌;β2-GP可抑制VEGFR1的表达,部分抑制p38MAPK磷酸化,从而抑制VEGF诱导的巨噬细胞迁移及趋化因子分泌。Objective To observe the chemotactic effects of vascular endothelial growth factor(VEGF)on macrophages and the effects of β2-glycoproteinⅠ(β2-GP)on the migration of macrophage induced by VEGF through RAW 264.7 incubation,and to discuss the possible mechanisms.Methods RAW 264.7 cells were incubated and divided into five groups:control group,VEGF group,VEGF+β2-GP group,VEGF+SB203580(p38 inhibitor)group,β2-GP gioup.Transwell cell migration assay was used to observe the chemotaxis of the cells in response to VEGF,and the effect of β2-GP on the macrophage migration induced by VEGF.ELISA was used to detect the excretion of monocyte chemotactic proteins-1(MCP-1),interleukin-8(IL-8)and mitochondrial intermediate peptidase-1β(M1P-1β).Western blotting was used to detect the expression of vascular endothelial growth factor receptor 1(VEGFR1)and activation of p38 mitogenactivated protein kinase(MAPK)signal transduction pathways.Results(1)Compared with control group,the number of macrophage migration in VEGF group increased(t=22.089,P<0.05).Compared with VEGF group,the macrophage migration in VEGF+β2-GP group and VEGF 4-SB203580 group was significantly reduced(t=22.687,63.625,all P<0.05).(2)Compared with control group,the secretion of macrophage chemokines MCP-1,IL-8 and MIP-1 increased in VEGF group(t=3.339,11.140,2.254,all P<0.05),and the secretion of the three chemokines induced by inhibiting VEGF decreased in VEGF+β2-GP group and VEGF+SB203580 group(t=3.148,2.402,2.798,6.754,4.459,5.528,all P<0.05).(3)Compared with control group,VEGFR1 expression and p38MAPK pathway phosphorylation were increased in VEGF group(t=5.161,P<0.05),while VEGFR1 expression and p38MAPK pathway phosphorylation were decreased in VEGF+β2-GP group(t=4.965,P<0.05).Conclusions VEGF can induce the migration of macrophages and secretion of chemokines MCP-1,IL-8 and MIP-1β through the increase of VEGFR1 on the macrophages and activate the p38MAPK transduction pathway;β2-GP can inhibit the expression of VEGFR1 and partially inhibit the phosph

关 键 词：β2-糖蛋白Ⅰ 血管内皮生长因子 巨噬细胞 糖尿病 P38丝裂原活化蛋白激酶 

分 类 号：R587.1[医药卫生—内分泌]