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C5a promotes migration, proliferation, and vessel formation in endothelial cells.

Kurihara R, Yamaoka K, Sawamukai N, Shimajiri S, Oshita K, Yukawa S, Tokunaga M, Iwata S, Saito K, Chiba K, Tanaka Y - Inflamm. Res. (2010)

Bottom Line: The goal of this paper is to investigate the effects of activated complement C5a on vascular endothelium during vessel formation.The actions were efficiently inhibited by a specific antagonist against C5a.Our results implicated C5a in vessel formation and as a potent target for management of inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: The First Department of Internal Medicine, University of Occupational and Environmental Health, Japan, Kitakyushu, Japan.

ABSTRACT

Objectives: The goal of this paper is to investigate the effects of activated complement C5a on vascular endothelium during vessel formation.

Methods: A human microvascular endothelial cell line (HMEC-1) derived from post-capillary venules in skin was used to measure DNA synthesis, proliferation and cell-cycle progression. In vitro ring-shaped formation by the cells was assessed by using type I collagen gel matrix and a cell-migration assay using the Chemotaxicell chamber. A Matrigel plug assay was performed to confirm the effect of C5a in vivo.

Results: C5a progressed the cell cycle of HMEC-1 into G2/M phases, and induced DNA synthesis and proliferation in a dose-dependent manner. C5a efficiently induced migration and ring-shaped structure formation both in vitro and in vivo. Furthermore, a C5a receptor antagonist (W-54011) suppressed all HMEC-1 activities including proliferation and migration.

Conclusions: Proliferation, migration, and ring-shaped formation by HMEC-1 cells was induced by C5a. The actions were efficiently inhibited by a specific antagonist against C5a. Our results implicated C5a in vessel formation and as a potent target for management of inflammatory diseases.

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Cell proliferation induced by C5a. [3H]-thymidine incorporation was measured with a liquid scintillation counter 48 h after adding the various concentrations of C5a, C5, or C3a to the HMEC-1 suspension. a C5a increased HMEC-1 proliferation in a dose-dependent manner, whereas C5 (b) and C3a (c) did not induce proliferation. d HMEC-1 was stimulated with either 10 nM C5a in the presence of various concentrations of C5a receptor antagonist W-54011 (closed column) or W-54011 alone (open column). The addition of W-54011 inhibited cell proliferation induced by C5a 10 nM in a dose-dependent manner, whereas W-54011 alone had no effect. Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test
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Fig2: Cell proliferation induced by C5a. [3H]-thymidine incorporation was measured with a liquid scintillation counter 48 h after adding the various concentrations of C5a, C5, or C3a to the HMEC-1 suspension. a C5a increased HMEC-1 proliferation in a dose-dependent manner, whereas C5 (b) and C3a (c) did not induce proliferation. d HMEC-1 was stimulated with either 10 nM C5a in the presence of various concentrations of C5a receptor antagonist W-54011 (closed column) or W-54011 alone (open column). The addition of W-54011 inhibited cell proliferation induced by C5a 10 nM in a dose-dependent manner, whereas W-54011 alone had no effect. Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test

Mentions: First, we used flow cytometric analysis to confirm that our HMEC-1 endothelial cell line expressed the C5a receptor (CD88) as expected (Fig. 1). Once established, the cells were stimulated with different concentrations of C5a and analyzed by [3H]-thymidine incorporation to assess cell proliferation. C5a induced a significant proliferation of HMEC-1 in a concentration-dependent manner, whereas other complement factors, C5 and C3a, had no such effect (Fig. 2a, b, c). The addition of W-54011, a specific C5a-receptor antagonist, significantly inhibited C5a-induced proliferation in a dose-dependent manner, whereas W-54011 alone had no effect (Fig. 2d).Fig. 1


C5a promotes migration, proliferation, and vessel formation in endothelial cells.

Kurihara R, Yamaoka K, Sawamukai N, Shimajiri S, Oshita K, Yukawa S, Tokunaga M, Iwata S, Saito K, Chiba K, Tanaka Y - Inflamm. Res. (2010)

Cell proliferation induced by C5a. [3H]-thymidine incorporation was measured with a liquid scintillation counter 48 h after adding the various concentrations of C5a, C5, or C3a to the HMEC-1 suspension. a C5a increased HMEC-1 proliferation in a dose-dependent manner, whereas C5 (b) and C3a (c) did not induce proliferation. d HMEC-1 was stimulated with either 10 nM C5a in the presence of various concentrations of C5a receptor antagonist W-54011 (closed column) or W-54011 alone (open column). The addition of W-54011 inhibited cell proliferation induced by C5a 10 nM in a dose-dependent manner, whereas W-54011 alone had no effect. Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test
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Related In: Results  -  Collection

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Fig2: Cell proliferation induced by C5a. [3H]-thymidine incorporation was measured with a liquid scintillation counter 48 h after adding the various concentrations of C5a, C5, or C3a to the HMEC-1 suspension. a C5a increased HMEC-1 proliferation in a dose-dependent manner, whereas C5 (b) and C3a (c) did not induce proliferation. d HMEC-1 was stimulated with either 10 nM C5a in the presence of various concentrations of C5a receptor antagonist W-54011 (closed column) or W-54011 alone (open column). The addition of W-54011 inhibited cell proliferation induced by C5a 10 nM in a dose-dependent manner, whereas W-54011 alone had no effect. Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test
Mentions: First, we used flow cytometric analysis to confirm that our HMEC-1 endothelial cell line expressed the C5a receptor (CD88) as expected (Fig. 1). Once established, the cells were stimulated with different concentrations of C5a and analyzed by [3H]-thymidine incorporation to assess cell proliferation. C5a induced a significant proliferation of HMEC-1 in a concentration-dependent manner, whereas other complement factors, C5 and C3a, had no such effect (Fig. 2a, b, c). The addition of W-54011, a specific C5a-receptor antagonist, significantly inhibited C5a-induced proliferation in a dose-dependent manner, whereas W-54011 alone had no effect (Fig. 2d).Fig. 1

Bottom Line: The goal of this paper is to investigate the effects of activated complement C5a on vascular endothelium during vessel formation.The actions were efficiently inhibited by a specific antagonist against C5a.Our results implicated C5a in vessel formation and as a potent target for management of inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: The First Department of Internal Medicine, University of Occupational and Environmental Health, Japan, Kitakyushu, Japan.

ABSTRACT

Objectives: The goal of this paper is to investigate the effects of activated complement C5a on vascular endothelium during vessel formation.

Methods: A human microvascular endothelial cell line (HMEC-1) derived from post-capillary venules in skin was used to measure DNA synthesis, proliferation and cell-cycle progression. In vitro ring-shaped formation by the cells was assessed by using type I collagen gel matrix and a cell-migration assay using the Chemotaxicell chamber. A Matrigel plug assay was performed to confirm the effect of C5a in vivo.

Results: C5a progressed the cell cycle of HMEC-1 into G2/M phases, and induced DNA synthesis and proliferation in a dose-dependent manner. C5a efficiently induced migration and ring-shaped structure formation both in vitro and in vivo. Furthermore, a C5a receptor antagonist (W-54011) suppressed all HMEC-1 activities including proliferation and migration.

Conclusions: Proliferation, migration, and ring-shaped formation by HMEC-1 cells was induced by C5a. The actions were efficiently inhibited by a specific antagonist against C5a. Our results implicated C5a in vessel formation and as a potent target for management of inflammatory diseases.

Show MeSH
Related in: MedlinePlus