title: Heme changes HIF-α, eNOS and nitrite production in HUVECs after simvastatin, HU, and ascorbic acid therapies.
abstract: The sickle cell disease (SCD) is a hemolytic genetic anemia characterized by free heme and hemoglobin release into intravascular spaces, with endothelial activation. Heme is a proinflammatory molecule able to directly activate vascular endothelium, thus, endothelial dysfunction and vascular disease are major chronic events described in SCD. The aim of this study was to evaluate the production of endothelial nitric oxide synthase (eNOS), nitrite and hypoxia inducible factor alpha (HIF-α) in HUVECs (human umbilical vein endothelial cells) activated by heme in response to simvastatin, hydroxyurea (HU), and ascorbic acid therapies. eNOS and HIF-α production were evaluated by ELISA and nitrite was measured by the Griess technique. The production of HIF-α increased when the cells were stimulated by heme (p<0.01), while treatment with HU and simvastatin reduced the production (p<0.01), and treatment with ascorbic acid increased HIF-1a production by the cells (p<0.01). Heme increased eNOS production, (p<0.01) but showed a heterogeneous pattern, and the lowest concentrations of all the treatments reduced the enzyme production (p<0.01). The nitrite production by HUVECs was enhanced by stimulation with heme (p<0.001) and was reduced by treatment with HU (p<0.001), ascorbic acid (p<0.001) and simvastatin (p<0.01). In summary, our results suggest that the hemolytic vascular microenvironment in SCD requires different therapeutic approaches to promote clinical improvement, and that a combination of therapies may be a viable strategy for treating patients.
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil. Electronic address: cguarda4@hotmail.com.
authors.forename: Caroline C
authors.initials: CC
authors.lastname: da Guarda
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil. Electronic address: rayrasantiago@hotmail.com.
authors.forename: Rayra P
authors.initials: RP
authors.lastname: Santiago
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil. Electronic address: pitanga.tn@hotmail.com.
authors.forename: Thassila N
authors.initials: TN
authors.lastname: Pitanga
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil. Electronic address: sanziobiomedico@hotmail.com.
authors.forename: Sanzio S
authors.initials: SS
authors.lastname: Santana
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil. Electronic address: dalila.zanette@bahia.fiocruz.br.
authors.forename: Dalila L
authors.initials: DL
authors.lastname: Zanette
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil. Electronic address: vborges@bahia.fiocruz.br.
authors.forename: Valéria M
authors.initials: VM
authors.lastname: Borges
authors.affiliation: Centro de Pesquisa Gonçalo Moniz/FIOCRUZ, Salvador, Brazil; Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Universidade Federal da Bahia, Salvador, Brazil. Electronic address: mari@bahia.fiocruz.br.
authors.forename: Marilda S
authors.initials: MS
authors.lastname: Goncalves
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood / Q000188:drug therapy* / Q000201:enzymology; D001205:Ascorbic Acid / Q000494:pharmacology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006418:Heme / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D009573:Nitrites / Q000378:metabolism*; D019821:Simvastatin / Q000494:pharmacology*
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009573:Nitrites; D006418:Heme; D019821:Simvastatin; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D001205:Ascorbic Acid; D006918:Hydroxyurea
keywords: Ascorbic acid; Endothelium; Heme; Hydroxyurea; Simvastatin