摘要
目的 探讨在血管内皮细胞参与血管形成过程中巨噬细胞所起的作用。方法 将血管内皮细胞 (ECV -3 0 4)接种培养 ,待细胞 60 %融合 ,按照不同的分组加入ECV -3 0 4,U93 7(巨细胞 1× 10 5/ml)和conA( 2 5ug/ml) ,共培养 48h后 ,用流式细胞仪检测细胞周期的变化 ;采用3 H -TdR掺入试验检测内皮细胞DNA合成变化 ;用RT -PCR技术检测内皮细胞VEGF受体KDRmRNA表达水平的变化 ;用免疫荧光在流式细胞仪上检测整和素受体ανβ3 表达的变化。结果 活化巨噬细胞使内皮细胞S期、DNA合成明显增加 (P <0 0 1) ,并使其VEGF受体KDRmRNA和整和素受体ανβ3 的表达水平明显上调 (P <0 0 1)。结论 活化的巨噬细胞通过影响内皮细胞的细胞周期、DNA合成、VEGF受体KDR及整和素受体ανβ3 的表达来促进内皮细胞的增殖、迁移及与基质的黏附 。
Objective To explore the role of active macrophages on the vascular endothelial participation in angiogenesis. Methods Vasculuar endothelial cell line ECV-304 was inoculated and cultured to 60% confluence.ECV-304 cells were then cultured lonely, with ConA(24μg/ml), with macrophage cell line U937(1×10 5/ml) or with U937 and ConA together to make four groups.The change of the ECV-304 cell cycle was determined by flow cytometry and that of the DNA synthesis was detected by 3H-TdR incorporation test. The change of the expression of endothelial VEGF receptor KDR mRNA was detected by RT-PCR while that of the integrin receptor α ν β 3 expression by immune fluorescent antibody technique on flow cytometry. Results Endothelial cells in S phase increased obviously with increased DNA synthesis in ECV-304 cells stimulated by active macrophages. In addition, the expression levels of the VEGF receptor KDR mRNA and integrin receptor α ν β 3 up-regulated significantly(P<0 01). Conclusions Active macrophage might modulate angiogenesis by influencing the cell cycle,DNA synthesis and the expression of VEGF receptor KDR mRNA and integrin receptor α ν β 3 of endothelia.
出处
《中国医师杂志》
CAS
2003年第11期1461-1463,共3页
Journal of Chinese Physician
基金
国家重点基础发展规划项目 (G1 9990 542 0 5)
