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

Park, Hyun-Joo; Kim, Su-Ryun; Choi, Yoon Kyung; Bae, Soo-Kyung; Bae, Moon‐Kyoung

doi:10.4196/kjpp.2015.19.4.327

Cited by 16 publications

(16 citation statements)

References 33 publications

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“…The existing literature on the role of orexin-A in the regulation of molecules implicated in the atherogenesis process is scarce. Recently, Kim et al found that incubation of HUVECs with orexin-A at concentrations of 0.1-0.3 µM and 0.2 µM, similar to those used in our study, for 2 and 8 h, respectively positively affect the expression and activity of HO-1, which is an anti-inflammatory molecule [11].…”

Section: Discussionsupporting

confidence: 76%

“…Recently, Kim et al demonstrated that orexin-A induced the expression of heme oxygenase-1 (HO-1) in endothelial cells (HUVECs) [11]. HO-1 is an anti-inflammatory, antioxidant, and cytoprotective enzyme that exerts beneficial effects on vascular damage and mediates the protection of endothelial cells by regulating endothelial cell activation and inhibiting endothelial cellular apoptosis [11].…”

Section: Introductionmentioning

confidence: 99%

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Orexin-A Exerts Equivocal Role in Atherosclerosis Process Depending on the Duration of Exposure: In Vitro Study

et al. 2019

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Orexin-A is a peptide hormone that plays a crucial role in feeding regulation and energy homeostasis. Diurnal intermittent fasting (DIF) has been found to increase orexin-A plasma levels during fasting hours, while Ramadan fasting which resembles DIF, has led to beneficial effects on endothelial function. Herein, we aimed to investigate the effects of orexin-A on the expression of molecules involved in the atherogenesis process: Monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) and tissue inhibitor of metalloproteinase-1 and 2 (TIMP-1 and TIMP-2), in human aortic endothelial cells (HAECs). HAECs were incubated with orexin-A at concentrations of 40 ng/mL, 200 ng/mL and 400 ng/mL for 6, 12 and 24 h. The mRNA levels of MCP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 and orexin-1 receptor were measured by real-time qPCR. We also evaluated the MMP-2, p38, phospho-p38, NF-κΒ/p65 as well as TIMP-1 protein levels by Western blot and ELISA, respectively. MMP-2 activity was measured by gelatin zymography. Short-term 6-h incubation of HAECs with orexin-A at a high concentration (400 ng/mL) decreased MCP-1, MMP-2 expression, MMP-2/TIMP-1 ratio (p < 0.05), and MMP-2 activity, while incubation for 24 h increased MCP-1, MMP-2 expression (p < 0.05), MMP-2/TIMP-1 and MMP-2/TIMP-2 ratio (p < 0.01 and p < 0.05, respectively) as well as MMP-2 activity. The dual effects of orexin-A are mediated, at least in part, via regulation of p38 and NF-κΒ pathway. Orexin-A may have an equivocal role in atherosclerosis process with its effects depending on the duration of exposure.

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Section: Discussionsupporting

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Orexin-A Exerts Equivocal Role in Atherosclerosis Process Depending on the Duration of Exposure: In Vitro Study

et al. 2019

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“…In patients with vascular disorders, their cerebrospinal fluids have reduced orexin A levels . In endothelial cells, orexin A induces angiogenesis and promotes the neovascularization . The vascular endothelial dysfunction is the early event of atherosclerosis.…”

Section: Introductionmentioning

confidence: 99%

Orexin A Suppresses Oxidized LDL Induced Endothelial Cell Inflammation via MAPK p38 and NF‐κB Signaling Pathway

Zhang

Liang

et al. 2018

IUBMB Life

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Orexin A is a multifaceted peptide produced in hypothalamus. We examined the effect of orexin A on vascular endothelial cells. Our study showed that orexin A had a profound inhibitory effect against endothelial inflammation by oxidized low-density lipoprotein (ox-LDL) in endothelial cells. Orexin A partially suppressed ox-LDL-induced monocytes THP-1 cells attachment to endothelial cells by limiting expression of vascular molecules including VCAM-1, ICAM-1, and E-selectin. Mechanistically, orexin A ameliorated endothelial dysfunction by reducing MAP kinase p38 and NF-κB activation via its receptor-OX1R. Orexin A suppressed phosphorylation of MAP kinase p38 and the NF-κB cascade kinases IKKα and IκBα, and prevented the shuttle of p65 protein into nuclear. Additionally, we reported that OX1R was expressed in HUVECs. Silence of OX1R completely abolished the inhibitory function of orexin in attachment of THP-1 cells. Collectively, our data suggest that orexin A ameliorated endothelial dysfunction under inflammatory stimuli. © 2018 IUBMB Life, 70(10):961-968, 2018.

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“…The capability of ECFCs to form tubule-like structures was determined using a tube formation assay with modifications of previous procedures [ 11 ]. Twenty-four-well plates were coated with ice-cold Matrigel solution (Phenol Red-Free; BD Biosciences, NJ, USA) and incubated at 37℃ for at least 30 min to allow the Matrigel to solidify.…”

Section: Methodsmentioning

confidence: 99%

Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

Lee

Kang

2018

Korean J Physiol Pharmacol

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The tube formation assay is a widely used in vitro experiment model to evaluate angiogenic properties by measuring the formation of tubular structures from vascular endothelial cells (ECs). in vitro experimental results are crucial when considered the advisability of moving forward to in vivo studies. Thus, the additional attentions to the in vitro assay is necessary to improve the quality of the pre-clinical data, leading to better decision-making for successful drug discovery. In this study, we improved the tube formation assay system in three aspects. First, we used human endothelial colony forming cells (ECFCs), which are endothelial precursors that have a robust proliferative capacity and more defined angiogenic characteristics compared to mature ECs. Second, we utilized a real-time cell recorder to track the progression of tube formation for 48 hours. Third, to minimize analysis error due to the limited observation area, we used image-stitching software to increase the microscope field of view to a 2×2 stitched area from the 4× object lens. Our advanced tube formation assay system successfully demonstrated the time-dependent dynamic progression of tube formation in the presence and absence of VEGF and FGF-2. Vatalanib, VEGF inhibitor, was tested by our assay system. Of note, IC50 values of vatalanib was different at each observation time point. Collectively, these results indicate that our advanced tube formation assay system replicates the dynamic progression of tube formation in response to angiogenic modulators. Therefore, this new system provides a sensitive and versatile assay model for evaluating pro- or anti-angiogenic drugs.

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

Cited by 16 publications

References 33 publications

Orexin-A Exerts Equivocal Role in Atherosclerosis Process Depending on the Duration of Exposure: In Vitro Study

Orexin-A Exerts Equivocal Role in Atherosclerosis Process Depending on the Duration of Exposure: In Vitro Study

Orexin A Suppresses Oxidized LDL Induced Endothelial Cell Inflammation via MAPK p38 and NF‐κB Signaling Pathway

Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

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