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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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Related in: MedlinePlus

Cell viability was measured using the Tetra Color One assay. The optical density of each well was measured at 450 nm. Phosphate-buffered saline (PBS) alone was added to control samples. a C5a increased the number of viable HMEC-1 cells at 1 and 10 nM, while C5 (b) and C3a (c) had no such effects. d W-54011 inhibited the C5a-augmented viable cell count (closed bars), but had no effect when added alone (open bars). Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test
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Fig4: Cell viability was measured using the Tetra Color One assay. The optical density of each well was measured at 450 nm. Phosphate-buffered saline (PBS) alone was added to control samples. a C5a increased the number of viable HMEC-1 cells at 1 and 10 nM, while C5 (b) and C3a (c) had no such effects. d W-54011 inhibited the C5a-augmented viable cell count (closed bars), but had no effect when added alone (open bars). Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test

Mentions: The effect of C5a on cell viability was then evaluated using the Tetra Color One assay with absorbance measured at 450 nm. C5a at both 1 and 10 nM increased significantly the HMEC-1 viable cell counts, whereas C5 and C3a did not affect HMEC-1 viability, even at 10 nM (Fig. 4a, b, c). The addition of 1 or 10 nM W-54011 suppressed the proliferative effects of 10 nM C5a. Importantly, W-54011 alone had no effect on viable cell counts, confirming its low toxicity (Fig. 4d).Fig. 4


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 viability was measured using the Tetra Color One assay. The optical density of each well was measured at 450 nm. Phosphate-buffered saline (PBS) alone was added to control samples. a C5a increased the number of viable HMEC-1 cells at 1 and 10 nM, while C5 (b) and C3a (c) had no such effects. d W-54011 inhibited the C5a-augmented viable cell count (closed bars), but had no effect when added alone (open bars). Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2902742&req=5

Fig4: Cell viability was measured using the Tetra Color One assay. The optical density of each well was measured at 450 nm. Phosphate-buffered saline (PBS) alone was added to control samples. a C5a increased the number of viable HMEC-1 cells at 1 and 10 nM, while C5 (b) and C3a (c) had no such effects. d W-54011 inhibited the C5a-augmented viable cell count (closed bars), but had no effect when added alone (open bars). Bars represent mean ± SEM (n = 5). **P < 0.01 by Dunnett’s multiple comparison test
Mentions: The effect of C5a on cell viability was then evaluated using the Tetra Color One assay with absorbance measured at 450 nm. C5a at both 1 and 10 nM increased significantly the HMEC-1 viable cell counts, whereas C5 and C3a did not affect HMEC-1 viability, even at 10 nM (Fig. 4a, b, c). The addition of 1 or 10 nM W-54011 suppressed the proliferative effects of 10 nM C5a. Importantly, W-54011 alone had no effect on viable cell counts, confirming its low toxicity (Fig. 4d).Fig. 4

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