caa a22 4500 44531432X CHVBK 20180317142641.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s11095-011-0528-0 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0528-0 Mitochondrial Antioxidants Alleviate Oxidative and Nitrosative Stress in a Cellular Model of Sepsis [Elektronische Daten] [Nadezda Apostolova, Remedios Garcia-Bou, Antonio Hernandez-Mijares, Raul Herance, Milagros Rocha, Victor Victor] ABSTRACT: Purpose: Mitochondrial dysfunction plays a key role in sepsis. Methods: We used a sepsis model of human endothelial cells (HUVEC) to study mitochondrial function during normoxic (21% O2) and hypoxic (1% O2) conditions. Results: When stimulated with a LPS cocktail, HUVEC displayed an increase of nitric oxide (NO) in normoxic and hipoxic conditions, being higher at 21% O2. LPS-activation for 24h at 1% O2 increased ROS production, which was reversed with the mitochondrial antioxidant Mitoquinone (MQ) and Glutathione Ethyl Ester (GEE). Activated cells displayed diminished mitochondrial O2 consumption with specific inhibition of Complex I, accompanied by increase in tyrosine nitration and Type II NOS protein expression, effects which were recovered by antioxidants and/or with L-NAME. These parameters varied with O2 environment, namely inhibition of respiration observed in both O2 environments at 24h was very similar, whereas O2 consumption rate fell earlier in 1% O2-exposed cells. While no significant differences were detected at earlier time points, at 24h tyrosine nitration was higher in normoxic vs. hypoxic cells. Conclusions: Mitochondria are heavily implicated in sepsis. Mitochondrial antioxidants provide a mechanistic model for the development of potential therapies. Springer Science+Business Media, LLC, 2011 hypoxia nationallicence mitochondria nationallicence mitoquinone nationallicence reactive oxygen species nationallicence sepsis nationallicence DAF-FM : 4-amino-5-methylamino-2′,7′-difluorofluorescein nationallicence DCFH-DA : 2′,7′-dichlorodihydrofluorescein diacetate nationallicence DHR123 : dihydrorhodamine 123 nationallicence GEE : glutathione ethyl ester nationallicence HBSS : Hank's balanced salt solution nationallicence HUVEC : human umbilical vein endothelial cells nationallicence L-NAME : L-NG-nitroarginine methyl ester nationallicence MCB : monochlorobimane nationallicence MOF : multiple organ failure nationallicence MQ : mitoquinone nationallicence PBS : phosphate-buffered saline nationallicence ROS : reactive oxygen species nationallicence TPP : triphenylphosphonium nationallicence Type II NOS : inducible nitric oxide synthase nationallicence Apostolova Nadezda Department of Pharmacology, University of Valencia, Valencia, Spain aut Garcia-Bou Remedios University Hospital Doctor Peset Foundation, Avda Gaspar Aguilar 90, 46017, Valencia, Spain aut Hernandez-Mijares Antonio Endocrinology Service, University Hospital Doctor Peset, Valencia, Spain aut Herance Raul High Technology Institute-PRBB, Barcelona, Spain aut Rocha Milagros Department of Pharmacology, University of Valencia, Valencia, Spain aut Victor Victor Department of Pharmacology, University of Valencia, Valencia, Spain aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/11(2011-11-01), 2910-2919 0724-8741 28:11<2910 2011 28 11095 https://doi.org/10.1007/s11095-011-0528-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0528-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Apostolova Nadezda Department of Pharmacology, University of Valencia, Valencia, Spain aut NATIONALLICENCE 700 1- Garcia-Bou Remedios University Hospital Doctor Peset Foundation, Avda Gaspar Aguilar 90, 46017, Valencia, Spain aut NATIONALLICENCE 700 1- Hernandez-Mijares Antonio Endocrinology Service, University Hospital Doctor Peset, Valencia, Spain aut NATIONALLICENCE 700 1- Herance Raul High Technology Institute-PRBB, Barcelona, Spain aut NATIONALLICENCE 700 1- Rocha Milagros Department of Pharmacology, University of Valencia, Valencia, Spain aut NATIONALLICENCE 700 1- Victor Victor Department of Pharmacology, University of Valencia, Valencia, Spain aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/11(2011-11-01), 2910-2919 0724-8741 28:11<2910 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer