scholarly journals Measurement of cellular oxygen consumption and extracellular acidification v2 (protocols.io.v92e98e)

protocols.io ◽  
2018 ◽  
Author(s):  
Kiichi Hirota ◽  
Yoshiyuki Matsuo
Keyword(s):  
Oxygen Consumption ◽  
Extracellular Acidification ◽  
Cellular Oxygen

scholarly journals Cytotoxicity and Mitochondrial Effects of Phenolic and Quinone-Based Mitochondria-Targeted and Untargeted Antioxidants on Human Neuronal and Hepatic Cell Lines: A Comparative Analysis

Biomolecules ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1605
Author(s):  
Carlos Fernandes ◽  
Afonso J. C. Videira ◽  
Caroline D. Veloso ◽  
Sofia Benfeito ◽  
Pedro Soares ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Lines ◽  
Mitochondrial Membrane ◽  
Hepatic Cell ◽  
Therapeutic Application ◽  
Extracellular Acidification ◽  
Mitochondrial Oxidative Stress ◽  
Drug Candidates ◽  
Cellular Oxygen ◽  
Hepatic Cell Lines

Mitochondriotropic antioxidants (MC3, MC6.2, MC4 and MC7.2) based on dietary antioxidants and analogs (caffeic, hydrocaffeic, trihydroxyphenylpropanoic and trihydroxycinnamic acids) were developed. In this study, we evaluate and compare the cytotoxicity profile of novel mitochondria-targeted molecules (generally known as MitoCINs) on human HepG2 and differentiated SH-SY5Y cells with the quinone-based mitochondria-targeted antioxidants MitoQ and SkQ1 and with two non-targeted antioxidants, resveratrol and coenzyme Q10 (CoQ10). We further evaluate their effects on mitochondrial membrane potential, cellular oxygen consumption and extracellular acidification rates. Overall, MitoCINs derivatives reduced cell viability at concentrations about six times higher than those observed with MitoQ and SkQ1. A toxicity ranking for both cell lines was produced: MC4 < MC7.2 < MC3 < MC6.2. These results suggest that C-6 carbon linker and the presence of a pyrogallol group result in lower cytotoxicity. MC3 and MC6.2 affected the mitochondrial function more significantly relative to MitoQ, SkQ1, resveratrol and CoQ10, while MC4 and MC7.2 displayed around 100–1000× less cytotoxicity than SkQ1 and MitoQ. Based on the mitochondrial and cytotoxicity cellular data, MC4 and MC7.2 are proposed as leads that can be optimized to develop safe drug candidates with therapeutic application in mitochondrial oxidative stress-related diseases.

Nitric oxide alters metabolism in isolated alveolar type II cells

1996 ◽  
Vol 271 (1) ◽  
pp. L23-L30 ◽  
Author(s):  
P. R. Miles ◽  
L. Bowman ◽  
L. Huffman
Keyword(s):  
Nitric Oxide ◽  
Oxygen Consumption ◽  
Atp Synthesis ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Atp Content ◽  
Alveolar Type Ii Cells ◽  
Type Ii Cell ◽  
Cellular Oxygen

Alveolar type II cells may be exposed to nitric oxide (.NO) from external sources, and these cells can also generate .NO. Therefore we studied the effects of altering .NO levels on various type II cell metabolic processes. Incubation of cells with the .NO generator, S-nitroso-N-acetylpenicillamine (SNAP; 1 mM), leads to reductions of 60-70% in the synthesis of disaturated phosphatidylcholines (DSPC) and cell ATP levels. Cellular oxygen consumption, an indirect measure of cell ATP synthesis, is also reduced by SNAP. There is no direct effect of SNAP on lung mitochondrial ATP synthesis, suggesting that .NO does not directly inhibit this process. On the other hand, incubation of cells with NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase (NOS), the enzyme responsible for .NO synthesis, results in increases in DSPC synthesis, cell ATP content, and cellular oxygen consumption. The L-NAME effects are reversed by addition of L-arginine, the substrate for NOS. Production of .NO by type II cells is inhibited by L-NAME, a better inhibitor of constitutive NOS (cNOS) than inducible NOS (iNOS), and is reduced in the absence of external calcium. Aminoguanidine, a specific inhibitor of iNOS, has no effect on cell ATP content or on .NO production. These results indicate that alveolar type II cell lipid and energy metabolism can be affected by .NO and suggest that there may be cNOS activity in these cells.

scholarly journals Evidence for a Physiological Mitochondrial Angiotensin II System in the Kidney Proximal Tubules

Hypertension ◽  
2020 ◽  
Vol 76 (1) ◽  
pp. 121-132
Author(s):  
Xiao Chun Li ◽  
Xinchun Zhou ◽  
Jia Long Zhuo
Keyword(s):  
Blood Pressure ◽  
Oxygen Consumption ◽  
Angiotensin Ii ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Physiological Role ◽  
Ang Ii ◽  
Extracellular Acidification ◽  
Extracellular Acidification Rate ◽  
Mitochondria Targeting

The present study tested the hypotheses that overexpression of an intracellular Ang II (angiotensin II) fusion protein, mito-ECFP/Ang II, selectively in the mitochondria of mouse proximal tubule cells induces mitochondrial oxidative and glycolytic responses and elevates blood pressure via the Ang II/AT 1a receptor/superoxide/NHE3 (the Na + /H + exchanger 3)-dependent mechanisms. A PT-selective, mitochondria-targeting adenoviral construct encoding Ad-sglt2-mito-ECFP/Ang II was used to test the hypotheses. The expression of mito-ECFP/Ang II was colocalized primarily with Mito-Tracker Red FM in mouse PT cells or with TMRM in kidney PTs. Mito-ECFP/Ang II markedly increased oxygen consumption rate as an index of mitochondrial oxidative response (69.5%; P <0.01) and extracellular acidification rate as an index of mitochondrial glycolytic response (34%; P <0.01). The mito-ECFP/Ang II–induced oxygen consumption rate and extracellular acidification rate responses were blocked by AT 1 blocker losartan ( P <0.01) and a mitochondria-targeting superoxide scavenger mito-TEMPO ( P <0.01). By contrast, the nonselective NO inhibitor L-NAME alone increased, whereas the mitochondria-targeting expression of AT 2 receptors (mito-AT 2 /GFP) attenuated the effects of mito-ECFP/Ang II ( P <0.01). In the kidney, overexpression of mito-ECFP/Ang II in the mitochondria of the PTs increased systolic blood pressure 12±3 mm Hg ( P <0.01), and the response was attenuated in PT-specific PT- Agtr1a −/− and PT- Nhe3 −/− mice ( P <0.01). Conversely, overexpression of AT 2 receptors selectively in the mitochondria of the PTs induced natriuretic responses in PT- Agtr1a −/− and PT- Nhe3 −/− mice ( P <0.01). Taken together, these results provide new evidence for a physiological role of PT mitochondrial Ang II/AT 1a /superoxide/NHE3 and Ang II/AT 2 /NO/NHE3 signaling pathways in maintaining blood pressure homeostasis.

Effects of Bupivacaine on Cellular Oxygen Consumption and Adenine Nucleotide Metabolism

1994 ◽  
Vol 78 (2) ◽  
pp. 335-339 ◽  
Author(s):  
François Sztark ◽  
Olivier Tueux ◽  
Philippe Erny ◽  
Philippe Dabadie ◽  
Jean-Pierre Mazat
Keyword(s):  
Oxygen Consumption ◽  
Adenine Nucleotide ◽  
Nucleotide Metabolism ◽  
Adenine Nucleotide Metabolism ◽  
Cellular Oxygen

Modification of the Cytosensor™ microphysiometer to simultaneously measure extracellular acidification and oxygen consumption rates

2003 ◽  
Vol 496 (1-2) ◽  
pp. 93-101 ◽  
Author(s):  
Sven E. Eklund ◽  
David E. Cliffel ◽  
Eugene Kozlov ◽  
Ales Prokop ◽  
John Wikswo ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Extracellular Acidification ◽  
Consumption Rates

scholarly journals Macrophages from the upper and lower human respiratory tract are metabolically distinct

2018 ◽  
Vol 315 (5) ◽  
pp. L752-L764 ◽  
Author(s):  
Katelyn S. Lavrich ◽  
Adam M. Speen ◽  
Andrew J. Ghio ◽  
Philip A. Bromberg ◽  
James M. Samet ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Respiratory Tract ◽  
Mitochondrial Respiration ◽  
Ex Vivo ◽  
Air Pollutant ◽  
Induced Sputum ◽  
Blood Monocytes ◽  
Extracellular Acidification ◽  
Healthy Human ◽  
Lung Macrophages

The function and cell surface phenotype of lung macrophages vary within the respiratory tract. Alterations in the bioenergetic profile of macrophages may also be influenced by their location within the respiratory tract. This study sought to characterize the bioenergetic profile of macrophages sampled from different locations within the respiratory tract at baseline and in response to ex vivo xenobiotic challenge. Surface macrophages recovered from healthy volunteers by induced sputum and by bronchial and bronchoalveolar lavage were profiled using extracellular flux analyses. Oxygen consumption and extracellular acidification rates were measured at rest and after stimulation with lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA), or 1,2-naphthoquinone (1,2-NQ). Oxygen consumption and extracellular acidification rates were highly correlated for all macrophage samples. Induced sputum macrophages had relatively higher oxygen consumption and extracellular acidification rates and were largely reliant on glycolysis. In contrast, bronchial fraction and bronchoalveolar macrophages depended more heavily on mitochondrial respiration. Bronchoalveolar macrophages showed elevated LPS-induced cytokine responses. Unlike their autologous peripheral blood monocytes, lung macrophages from any source did not display bioenergetic changes following LPS stimulation. The protein kinase C activator PMA did not affect mitochondrial respiration, whereas the air pollutant 1,2-NQ induced marked mitochondrial dysfunction in bronchoalveolar and bronchial fraction macrophages. The bioenergetic characteristics of macrophages from healthy individuals are dependent on their location within the respiratory tract. These findings establish a regional bioenergetic profile for macrophages from healthy human airways that serves as a reference for changes that occur in disease.

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