a811e1e926427b75e75f1d137923be4e ijtm-01-00022.pdf 7bac592ee4a13ed8f11065c7e55811c2effcc5ef ijtm-01-00022.pdf 8013362bf29a83ab4a4add7156e79715575b97ed5bab2da4495a3bcac884ee45 ijtm-01-00022.pdf Title: Hypoxia Increases Nitric Oxide-Dependent Inhibition of Angiogenic Growth Subject: Nitric oxide (NO) is a proangiogenic factor acting through the soluble guanylate cyclase (sGC) pathway. However, angiogenic growth increases energy demand, which may be hampered by NO inhibition of cytochrome c oxidase (CcO). Then, NO activity would be the balanced result of sGC activation (pro-angiogenic) and CcO inhibition (anti-angiogenic). NO activity in a rat and eNOS-/- mice aortic ring angiogenic model and in a tube formation assay (human aortic endothelial cells) were analyzed in parallel with mitochondrial O2 consumption. Studies were performed with NO donor (DETA-NO), sGC inhibitor (ODQ), and NOS or nNOS inhibitors (L-NAME or SMTC, respectively). Experiments were performed under different O2 concentrations (0–21%). Key findings were: (i) eNOS-derived NO inhibits angiogenic growth by a mechanism independent on sGC pathway and related to inhibition of mitochondrial O2 consumption; (ii) NO inhibition of the angiogenic growth is more evident in hypoxic vessels; (iii) in the absence of eNOS-derived NO, the modulation of angiogenic growth, related to hypoxia, disappears. Therefore, NO, but not lower O2 levels, decreases the angiogenic response in hypoxia through competitive inhibition of CcO. This anti-angiogenic activity could be a promising target to impair pathological angiogenesis in hypoxic conditions, as it occurs in tumors or ischemic diseases. Keywords: nitric oxide; hypoxia; angiogenesis; mitochondrial O2 consumption Author: Cristina Arce, Diana Vicente, Fermí Monto, Laura González, Cristina Nuñez, Víctor M. Victor, Francesc Jiménez-Altayó and Pilar D’Ocon Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Dec 8 11:02:10 2021 CET ModDate: Wed Dec 8 11:12:09 2021 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3040936 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ARIQJE+VnURWPalladioL Type 1 Custom yes yes no 10 0 MSBFSK+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 QBUKVH+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 HIDLLH+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 UWJUYN+CMSY10 Type 1 Builtin yes yes no 28 0 JYPLJJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 59 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 63 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 67 0 ZXDKIP+EURM10 Type 1 Builtin yes yes no 82 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 92 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 101 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 107 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 113 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 129 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 135 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 141 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 157 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 163 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 169 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 181 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 187 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 193 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 205 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 211 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 217 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 226 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 232 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 238 0 FNTSBS+ArialMT CID TrueType Identity-H yes yes yes 244 0 FNTSBS+Arial-BoldMT CID TrueType Identity-H yes yes yes 250 0 FNTSBS+MalgunGothic CID TrueType Identity-H yes yes yes 256 0 FNTSBS+Calibri CID TrueType Identity-H yes yes yes 262 0 CRUZTT+VnURWPalladioL-Italic Type 1 Custom yes yes no 288 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-02-26 02:49:47 CET RepresentationInformation: ijtm-01-00022.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-02-25 16:45:54 CET Size: 3040936 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 462 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Animals and Tissues Destination: subsection.2.1 Item: Title: Arterial Ring Model of Angiogenesis Destination: subsection.2.2 Item: Title: Tube Formation Assay Destination: subsection.2.3 Item: Title: Functional Experiments Destination: subsection.2.4 Item: Title: Oxygen Consumption Measurements Destination: subsection.2.5 Item: Title: qRT-PCR Destination: subsection.2.6 Item: Title: Statistical Analysis Destination: subsection.2.7 Item: Title: Results Destination: section.3 Children: Item: Title: The NO Donor DETA-NO Inhibits Angiogenic Growth by a Mechanism Independent of sGC Activation Destination: subsection.3.1 Item: Title: NO Endogenously Released by eNOS Inhibits Angiogenic Growth and O2 Consumption Destination: subsection.3.2 Item: Title: eNOS Expression Decreases in Sprouting Microvessels vs. the Aortic Ring Destination: subsection.3.3 Item: Title: NO Inhibition of O2 Consumption and Angiogenic Growth Is More Pronounced in Hypoxic Vessels Destination: subsection.3.4 Item: Title: Discussion Destination: section.4 Children: Item: Title: NO Inhibits Angiogenic Growth by a Mechanism Related to Inhibition of CcO and Independent of the sGC Pathway Destination: subsection.4.1 Item: Title: NO-Induced Inhibition of Angiogenic Growth and O2 Consumption Depends on the O2 Concentration and Is More Pronounced in Hypoxic Vessels Destination: subsection.4.2 Item: Title: References Destination: subsection.4.2 Info: Title: Hypoxia Increases Nitric Oxide-Dependent Inhibition of Angiogenic Growth Author: Cristina Arce, Diana Vicente, Fermí Monto, Laura González, Cristina Nuñez, Víctor M. Victor, Francesc Jiménez-Altayó and Pilar D’Ocon Subject: Nitric oxide (NO) is a proangiogenic factor acting through the soluble guanylate cyclase (sGC) pathway. However, angiogenic growth increases energy demand, which may be hampered by NO inhibition of cytochrome c oxidase (CcO). Then, NO activity would be the balanced result of sGC activation (pro-angiogenic) and CcO inhibition (anti-angiogenic). NO activity in a rat and eNOS-/- mice aortic ring angiogenic model and in a tube formation assay (human aortic endothelial cells) were analyzed in parallel with mitochondrial O2 consumption. Studies were performed with NO donor (DETA-NO), sGC inhibitor (ODQ), and NOS or nNOS inhibitors (L-NAME or SMTC, respectively). Experiments were performed under different O2 concentrations (0 21%). Key findings were: (i) eNOS-derived NO inhibits angiogenic growth by a mechanism independent on sGC pathway and related to inhibition of mitochondrial O2 consumption; (ii) NO inhibition of the angiogenic growth is more evident in hypoxic vessels; (iii) in the absence of eNOS-derived NO, the modulation of angiogenic growth, related to hypoxia, disappears. Therefore, NO, but not lower O2 levels, decreases the angiogenic response in hypoxia through competitive inhibition of CcO. This anti-angiogenic activity could be a promising target to impair pathological angiogenesis in hypoxic conditions, as it occurs in tumors or ischemic diseases. 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