巨噬细胞对内皮细胞系同源盒B2基因和血管内皮生长因子受体mRNA及整合素ανβ3表达的影响  

Influence of macrophages on the expression of vascular endothelial growth factor receptor mRNA, homeobox B2 mRNA, and integrin ανβ3 in vascular endothelial strain

作　　者：刘亮[1,5] 刘畅[2] 张晓启 明佳[1] 刘旭盛[1] 徐辉[4] 程天民[4] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所,创伤,烧伤与复合伤国家重点实验室,重庆400038 [2]南京军区总医院干部肿瘤科 [3]解放军第二三四医院烧伤科 [4]第三军医大学全军复合伤研究所,创伤,烧伤与复合伤国家重点实验室 [5]南方医科大学南方医院烧伤科

出　　处：《中华烧伤杂志》2005年第3期199-202,共4页Chinese Journal of Burns

基　　金：国家重点基础研究发展规划资助项目(G1999054205)

摘　　要：目的　体外观察巨噬细胞(M)对血管内皮细胞同源盒(HOX)B2基因mRNA和血管内皮生长因子(VEGF)受体KDRmRNA及整合素ανβ3表达的影响。　方法　体外培养人脐静脉血管内皮细胞系ECV304,分为(1)ECV304组; (2)ECV304 +伴刀豆球蛋白A(conA,终浓度25mg/L)组; (3)ECV304+人M系U937组:ECV304细胞中加入1×105 /mlU937细胞; (4)ECV304+U937+conA组:ECV304中加入1×105 /mlU937细胞及终浓度25mg/L的conA.反应48h后,用免疫荧光技术检测各组ECV304细胞整合素ανβ3的表达情况,并用逆转录聚合酶链反应(RT PCR)技术检测细胞KDRmRNA和HOXB2基因mRNA的表达水平。　结果　ECV304组细胞的整合素ανβ3、KDRmRNA及HOXB2基因mRNA的表达水平分别为6. 7±1. 5、0. 633±0. 012、0. 674±0. 004。ECV304+U937+conA组细胞上述指标较ECV304组均明显上调(P<0. 01 ),分别为10. 2±1. 7、0. 879±0 003、0. 947±0 003。其余两组细胞上述指标与ECV304组比较,差异均无统计学意义(P>0. 05).结论　被conA活化的M通过影响内皮细胞的KDRmRNA、HOXB2mRNA基因及整合素ανβ3的表达,来促进内皮细胞增殖、迁移及与基质黏附,从而调节血管生成。Objective To investigate the influence of macrophages on the expression of the vascular endothelial growth factor(VEGF) receptor (KDR) mRNA, homeobox B2(HOXB2) mRNA, and integrin ανβ3 in vitro in vascular endothelial strain. Methods Human umbilical vein cells (ECV304) were cultured in vitro and divided into 4 groups, i.e.(1) ECV304 group, (2) ECV304+conA group [with conA (25 μg/ml in culture) added to ECV304], (3) ECV304+U937 group (with 1×10 5/ml of U937 cells added to 1×10 5/ml ECV 304), (4) ECV304+U937+conA group [with 1×10 5/ml of U937 cells and conA (25 μg/ml in culture)]groups. Forty-eight hours after culturing, the expression of integrin receptor ανβ3 and the changes in the expression of KDR mRNA and HOXB2 mRNA in each group were determined by immunofluorescent technique and RT-PCR, respectively. Results The expression of integrin receptor ανβ3, KDR mRNA, and HOXB2 mRNA in ECV304 group were 6.7±1.5, 0.633±0.012, and 0.674±0.004, respectively, while those in ECV304+U937+conA group (10.2±1.7, 0.879±0.003, 0.947±0.003) were obviously more upregulated when compared with those in ECV304 group( P <0.01). No difference in the above indices was found between ECV304 and ECV304+conA, ECV304+U937 groups ( P >0.05). Conclusion Macrophages activated by ConA can accelerate the proliferation, migration and adhesion to the basement membrane matrix of vascular endothelial cells through the influence on the expression of KDR mRNA, HOXB2 mRNA and integrin ανβ3, and through this pathway the angiogenesis is modulated.

关 键 词：整合素αvβ3 受体mRNA 同源盒 血管内皮生长因子(VEGF) B2基因 ECV304细胞 基因mRNA 逆转录聚合酶链反应 巨噬细胞(Mφ) U937细胞 HOXB2 血管内皮细胞系 伴刀豆球蛋白 conA活化 MRNA基因 内皮细胞增殖 技术检测 受体KDR 

分 类 号：R346[医药卫生—基础医学]