Angiogenic effect of intercellular adhesion molecule-1

Abstract

In order to investigate the angiogenic effect of intercellular adhesion molecule-1 (ICAM-1), two parts of experiment were performed. Chick embryo chorioallantoic membrane (CAM) assay was used for in vivo angiogenic research. The chick embryos were divided into 4 groups: ICAM-1 group (divided into 3 subgroups, I, II and III) for screening the angiogenic effect of ICAM-1 by adding different concentrations of ICAM-1 (0.1, 0.2 and 0.3 μg/μL) 5 μL into the chick embryo CAMs on the day 10 after incubation for every subgroup; Anti-ICAM-1 group A (divided into 2 subgroups, I and II) by adding different concentrations of Anti-ICAM-1 (1:100, 1:50) 5 μL into the chick embryo CAMs on the day 10 after incubation for every subgroup to evaluate the effect of ICAM-1 on the survival of microvessels through observing whether Anti-ICAM-1 could induce involution of the microvessels on CAMs; Anti-ICAM-1 group B (divided into 2 subgroups, I and II) by adding different concentrations of Anti-ICAM-1 (1:100, 1:50) 5 μL into the chick embryo CAMs on the day 6 after incubation for every subgroup to evaluate whether ICAM-1 involved in embryonic angiogenesis through observing the growth of microvessels on CAMs; Control group: ICAM-1 or Anti-ICAM-1 was substituted by PBS 5 μL on the day 10 or day 6 after incubation. Three days later, the CAMs were photographed in vivo, excised, sectioned and the number of microvessels was counted. In ICAM-1 group, there was increased number of microvessels arranged radially with “spoked-wheel” pattern around the gelatin sponges. The new microvessels growing perpendicularly to gelatin sponges were observed. The number of the microvessels growing in the CAM mesenchymes around the sponges in 3 subgroups was higher than that in control group (P<0.01), however, there was no significant difference among the 3 subgroups (P>0.05). In anti-ICAM-1 group A, the radially arranged microvessels were very unclear around the sponges contrast to that of ICAM-1 group. Few new microvessels were detected in the center of the sponges. The number of the microvessels growing in the CAM mesenchymes around the sponges in subgroup II was lower than that in control group (P<0.01). There was no significant difference in the number of the microvessels around the sponges between subgroup I and control group (P>0.05). In anti-ICAM-1 group B, the radially arranged microvessels were very unclear around the sponges contrast to that of control group. New microvessels were very scarce in the center of the sponges. The number of the microvessels growing in the CAM mesenchymes around the sponges in the 2 subgroups were less than that in control group (P<0.01), and there was significant difference between the 2 subgroups (P<0.05). It was suggested that ICAM-1 could induce angiogenesis and support the survival of microvessels, and ICAM-1 was involved in embryonic angiogenesis.

Keywords

ICAM-1 / angiogenesis / immunohistochemistry / chick embryo chorioallantoic membrane

References

[1] KimS., BellK., MousaS. A., et al.. Regulation of angiogenesis in vivo by ligation of integrin alpha5betal with the central cell-binding domain of fibronectin. Am J Pathol, 2000, 156(4): 1345-1362

[2] HarryC. L., ThomasP. P.. Transcriptional regulation of the intercellular adhesion molecule 1 gene by inflammatory cytokines in human endothelial cell. J Biol Chem, 1995, 1270: 933-938

[3] RisauW.. Mechanisms of angiogenesis. Nature, 1997, 386(6626): 671-674

[4] ConwayE. M., CollenD., CarmelietP.. Molecular mechanisms of blood vessel growth. Cardiovascular Res, 2001, 49(3): 507-521

[5] LossinskyA. S., ButtleK. F., PlutaR., et al.. Immunoultrastructural expression of intercellular adhesion molecle-1 in endothelial cell vesiculotubular structures and vesiculovacuolar organelles in blood brain barrier development and injury. Cell Tissue Res, 1999, 295(1): 77-78

[6] GhoY. S., KimP. N., LiH. C., et al.. Stimulation of tumor growth by human soluble intercellular adhesion molecule-1. Cancer Res, 2001, 61(10): 4253-4257

[7] ShengY. L., ZhengQ. X., ZhaoH., et al.. The observation of mast cell behavior in hemangioma and vascular malformations. Chin J Pathol (Chinese), 1994, 23(5): 311-319

[8] LiW. W., GraysonG., ForlkmanJ., et al.. Sustained release of endotoxin. Invest Ophthalmol Vis Sci, 1991, 32: 2906-2911

[9] MatsushitaS., NitandaT., FurukawaT., et al.. The effect of a thymidien phosphorylase inhibitor on angiogenesis and apoptosis in tumor. Cancer Res, 1999, 59(8): 1911-1916

[10] LichtenbergJ., HansenC. A., Skak-NielsenT., et al.. The rat subcutaneous air sac model: a new and simple method for in vivo screening of antiangiogenesis. Pharmac Toxic, 1997, 81(6): 280-284