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Involvement of Heme Oxygenase-1 in Orexin-A-induced Angiogenesis in Vascular Endothelial Cells.

Kim MK, Park HJ, Kim SR, Choi YK, Bae SK, Bae MK - Korean J. Physiol. Pharmacol. (2015)

Bottom Line: Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin-A-induced angiogenesis in vivo and ex vivo.Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1.Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 626-870, Korea.

ABSTRACT
The cytoprotective enzyme heme oxygenase-1 (HO-1) influences endothelial cell survival, proliferation, inflammatory response, and angiogenesis in response to various angiogenic stimuli. In this study, we investigate the involvement of HO-1 in the angiogenic activity of orexin-A. We showed that orexin-A stimulates expression and activity of HO-1 in human umbilical vein endothelial cells (HUVECs). Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin-A-induced angiogenesis in vivo and ex vivo. Orexin-A-stimulated endothelial tube formation and chemotactic activity were also blocked in SnPP-treated vascular endothelial cells. Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1. Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

No MeSH data available.


Related in: MedlinePlus

Involvement of HO-1 in orexin-A-induced angiogenesis in vivo. (A) Matrigel was treated with orexin-A (200 nM) or PBS in the presence or absence of SnPP (20 µM), and then subcutaneously injected into mice. After 7 days, the plugs were obtained from each mouse and photographed. (B) The level of vessel formation was quantified by measuring hemoglobin content. Each value represents the mean of at least three animals. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone. (C) Matrigel plugs were stained using H&E and photographed. (D) Endothelial cells recruited into Matrigel plugs were immunostained with PECAM-1 antibody and observed using fluorescence microscopy.
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Figure 2: Involvement of HO-1 in orexin-A-induced angiogenesis in vivo. (A) Matrigel was treated with orexin-A (200 nM) or PBS in the presence or absence of SnPP (20 µM), and then subcutaneously injected into mice. After 7 days, the plugs were obtained from each mouse and photographed. (B) The level of vessel formation was quantified by measuring hemoglobin content. Each value represents the mean of at least three animals. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone. (C) Matrigel plugs were stained using H&E and photographed. (D) Endothelial cells recruited into Matrigel plugs were immunostained with PECAM-1 antibody and observed using fluorescence microscopy.

Mentions: Previously, we have suggested that orexin-A exerts angiogenic properties both in vivo and ex vivo [6]. To further assess whether increased HO-1 expression and activity is required for orexin-A-induced angiogenesis, Matrigel containing orexin-A, with or without SnPP, was subcutaneously injected into male C57BL/6 mice. After 7 days, the Matrigel plugs from each mouse were collected and analyzed. As shown in Fig. 2A, Matrigel containing orexin-A showed a remarkably reduced angiogenic effect in the presence of SnPP when compared with Matrigel containing orexin-A alone. The degree of blood vessel formation in the Matrigel plug was confirmed by measuring hemoglobin content (Fig. 2B). In addition, histological analysis indicated that inhibition of HO-1 by SnPP resulted in decreased levels of orexin-A-induced infiltration of endothelial cells (Fig. 2C). Infiltrated cells were positive for the endothelial marker PECAM-1 (Fig. 2D). These data demonstrate that HO-1 activity is required for orexin-A-mediated angiogenesis in vivo.


Involvement of Heme Oxygenase-1 in Orexin-A-induced Angiogenesis in Vascular Endothelial Cells.

Kim MK, Park HJ, Kim SR, Choi YK, Bae SK, Bae MK - Korean J. Physiol. Pharmacol. (2015)

Involvement of HO-1 in orexin-A-induced angiogenesis in vivo. (A) Matrigel was treated with orexin-A (200 nM) or PBS in the presence or absence of SnPP (20 µM), and then subcutaneously injected into mice. After 7 days, the plugs were obtained from each mouse and photographed. (B) The level of vessel formation was quantified by measuring hemoglobin content. Each value represents the mean of at least three animals. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone. (C) Matrigel plugs were stained using H&E and photographed. (D) Endothelial cells recruited into Matrigel plugs were immunostained with PECAM-1 antibody and observed using fluorescence microscopy.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499644&req=5

Figure 2: Involvement of HO-1 in orexin-A-induced angiogenesis in vivo. (A) Matrigel was treated with orexin-A (200 nM) or PBS in the presence or absence of SnPP (20 µM), and then subcutaneously injected into mice. After 7 days, the plugs were obtained from each mouse and photographed. (B) The level of vessel formation was quantified by measuring hemoglobin content. Each value represents the mean of at least three animals. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone. (C) Matrigel plugs were stained using H&E and photographed. (D) Endothelial cells recruited into Matrigel plugs were immunostained with PECAM-1 antibody and observed using fluorescence microscopy.
Mentions: Previously, we have suggested that orexin-A exerts angiogenic properties both in vivo and ex vivo [6]. To further assess whether increased HO-1 expression and activity is required for orexin-A-induced angiogenesis, Matrigel containing orexin-A, with or without SnPP, was subcutaneously injected into male C57BL/6 mice. After 7 days, the Matrigel plugs from each mouse were collected and analyzed. As shown in Fig. 2A, Matrigel containing orexin-A showed a remarkably reduced angiogenic effect in the presence of SnPP when compared with Matrigel containing orexin-A alone. The degree of blood vessel formation in the Matrigel plug was confirmed by measuring hemoglobin content (Fig. 2B). In addition, histological analysis indicated that inhibition of HO-1 by SnPP resulted in decreased levels of orexin-A-induced infiltration of endothelial cells (Fig. 2C). Infiltrated cells were positive for the endothelial marker PECAM-1 (Fig. 2D). These data demonstrate that HO-1 activity is required for orexin-A-mediated angiogenesis in vivo.

Bottom Line: Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin-A-induced angiogenesis in vivo and ex vivo.Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1.Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Physiology, School of Dentistry, Pusan National University, Yangsan 626-870, Korea.

ABSTRACT
The cytoprotective enzyme heme oxygenase-1 (HO-1) influences endothelial cell survival, proliferation, inflammatory response, and angiogenesis in response to various angiogenic stimuli. In this study, we investigate the involvement of HO-1 in the angiogenic activity of orexin-A. We showed that orexin-A stimulates expression and activity of HO-1 in human umbilical vein endothelial cells (HUVECs). Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin-A-induced angiogenesis in vivo and ex vivo. Orexin-A-stimulated endothelial tube formation and chemotactic activity were also blocked in SnPP-treated vascular endothelial cells. Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1. Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

No MeSH data available.


Related in: MedlinePlus