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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

Upregulation of HO-1 expression and activity by orexin-A in vascular endothelial cells. (A and B) HUVECs were treated with orexin-A (200 nM) or phosphate buffered saline (PBS) for the various times and concentrations shown. Expression of HO-1 was determined by western blot using HO-1 and α-tubulin antibodies (loading control). (C and D) Cells were incubated with orexin-A (200 nM) or PBS for the indicated times. Expression of HO-1 and HO-2 mRNA was analyzed by real-time PCR. These data represent the mean±SE of three experiments. *p<0.05 vs. control. (E) HUVECs were pretreated with SnPP (50 µM) for 15 min, followed by treatment with orexin-A for 4 hr. HO-1 enzyme activity was measured as described in the Materials and Methods. Three independent experiments were performed. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone.
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Figure 1: Upregulation of HO-1 expression and activity by orexin-A in vascular endothelial cells. (A and B) HUVECs were treated with orexin-A (200 nM) or phosphate buffered saline (PBS) for the various times and concentrations shown. Expression of HO-1 was determined by western blot using HO-1 and α-tubulin antibodies (loading control). (C and D) Cells were incubated with orexin-A (200 nM) or PBS for the indicated times. Expression of HO-1 and HO-2 mRNA was analyzed by real-time PCR. These data represent the mean±SE of three experiments. *p<0.05 vs. control. (E) HUVECs were pretreated with SnPP (50 µM) for 15 min, followed by treatment with orexin-A for 4 hr. HO-1 enzyme activity was measured as described in the Materials and Methods. Three independent experiments were performed. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone.

Mentions: To examine whether orexin-A affects HO-1 expression in vascular endothelial cells, HUVECs were treated with orexin-A and whole cell lysates were analyzed by Western blot. As shown in Fig. 1A, a time-dependent increase in HO-1 protein expression was observed in endothelial cells after orexin-A treatment. Furthermore, orexin-A induced HO-1 expression in a dose-dependent manner (Fig. 1B). Real-time PCR demonstrated that expression of HO-1 mRNA was also induced by orexin-A (Fig. 1C). By contrast, there was no effect on HO-2 mRNA levels in HUVECs incubated with orexin-A (Fig. 1D). An increase in HO-1 enzyme activity was also observed after orexin-A treatment, as determined by in vitro bilirubin production, and this increase was effectively attenuated by SnPP, a HO-1 inhibitor (Fig. 1E). These results indicate that orexin-A induces HO-1 mRNA, protein, and enzyme activity in vascular endothelial cells.


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)

Upregulation of HO-1 expression and activity by orexin-A in vascular endothelial cells. (A and B) HUVECs were treated with orexin-A (200 nM) or phosphate buffered saline (PBS) for the various times and concentrations shown. Expression of HO-1 was determined by western blot using HO-1 and α-tubulin antibodies (loading control). (C and D) Cells were incubated with orexin-A (200 nM) or PBS for the indicated times. Expression of HO-1 and HO-2 mRNA was analyzed by real-time PCR. These data represent the mean±SE of three experiments. *p<0.05 vs. control. (E) HUVECs were pretreated with SnPP (50 µM) for 15 min, followed by treatment with orexin-A for 4 hr. HO-1 enzyme activity was measured as described in the Materials and Methods. Three independent experiments were performed. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 1: Upregulation of HO-1 expression and activity by orexin-A in vascular endothelial cells. (A and B) HUVECs were treated with orexin-A (200 nM) or phosphate buffered saline (PBS) for the various times and concentrations shown. Expression of HO-1 was determined by western blot using HO-1 and α-tubulin antibodies (loading control). (C and D) Cells were incubated with orexin-A (200 nM) or PBS for the indicated times. Expression of HO-1 and HO-2 mRNA was analyzed by real-time PCR. These data represent the mean±SE of three experiments. *p<0.05 vs. control. (E) HUVECs were pretreated with SnPP (50 µM) for 15 min, followed by treatment with orexin-A for 4 hr. HO-1 enzyme activity was measured as described in the Materials and Methods. Three independent experiments were performed. *p<0.05 vs. control; #p<0.05 vs. orexin-A alone.
Mentions: To examine whether orexin-A affects HO-1 expression in vascular endothelial cells, HUVECs were treated with orexin-A and whole cell lysates were analyzed by Western blot. As shown in Fig. 1A, a time-dependent increase in HO-1 protein expression was observed in endothelial cells after orexin-A treatment. Furthermore, orexin-A induced HO-1 expression in a dose-dependent manner (Fig. 1B). Real-time PCR demonstrated that expression of HO-1 mRNA was also induced by orexin-A (Fig. 1C). By contrast, there was no effect on HO-2 mRNA levels in HUVECs incubated with orexin-A (Fig. 1D). An increase in HO-1 enzyme activity was also observed after orexin-A treatment, as determined by in vitro bilirubin production, and this increase was effectively attenuated by SnPP, a HO-1 inhibitor (Fig. 1E). These results indicate that orexin-A induces HO-1 mRNA, protein, and enzyme activity in vascular endothelial cells.

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