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Lipopolysaccharide-induced alterations in oxygen consumption and radical generation in endothelial cells

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Abstract

Oxygen consumption rate (OCR) and generation of superoxide and nitric oxide (NO) in mouse aortic endothelial cells (MAECs) treated with lipopolysaccharide (LPS) were studied. The OCR was determined in cell suspensions at 37 °C by electron paramagnetic resonance (EPR) spectroscopy. LPS significantly altered the OCR in a dose and time-dependent fashion. The OCR was significantly elevated immediately following the treatment of MAECs with LPS (5 and 10 μg/ml) and NADPH (100 μM) whereas the same was depressed 1 h after exposure to similar conditions of incubation. Under similar experimental conditions, superoxide generation was also determined by EPR spectroscopy and cytochrome c reduction assays. A marginal increase in the superoxide production was observed when the cells were treated with LPS and NADPH alone whereas the same was further enhanced significantly when the cells were treated with LPS and NADPH together. The increase in oxygen consumption and superoxide production caused by LPS was inhibited by diphenyleneiodonium (DPI), suggesting the involvement of NAD(P)H oxidase. A significant increase in the NO production by MAECs was noticed 1 h after treatment with LPS and was inhibited by L-NAME, further suggesting the involvement of nitric oxide synthase (NOS). Thus, on a temporal scale, LPS-induced alterations in oxygen consumption by MAECs may be under the control of dual regulation by NAD(P)H oxidase and NOS. (Mol Cell Biochem 278: 119–127, 2005)

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Abbreviations

CHO:

Chinese hamster ovary

DMPO:

5,5-dimethyl-1-pyrroline-N-oxide

DPI:

diphenyleneiodonium

DTPA:

diethylenetriaminepentaacetic acid

EPR:

electron paramagnetic resonance

LiNc-BuO:

lithium octa-N-butoxy-naphthalocyanine

L-NAME:

NG-nitro-L-arginine methyl ester

LPS:

lipopolysaccharide

MAECs:

mouse aortic endothelial cells

NO:

nitric oxide

NOS:

nitric oxide synthase

OCR:

oxygen consumption rate

ROS:

reactive oxygen species

SNAP:

-nitroso-N-acetylpenicillamine

SOD:

superoxide dismutase

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Authors and Affiliations

  1. Center for Biomedical EPR Spectroscopy and Imaging, Davis Heart and Lung Research Institute, Department of Internal Medicine, The Ohio State University, Columbus, OH, 43210, USA

    Ramasamy P. Pandian, Vijay Kumar Kutala, Alex Liaugminas, Narasimham L. Parinandi & Periannan Kuppusamy

  2. Davis Heart and Lung Research Institute, The Ohio State University, 420 West 12th Ave, Room 114, Columbus, OH, 43210, USA

    Periannan Kuppusamy

Authors
  1. Ramasamy P. Pandian
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  2. Vijay Kumar Kutala
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  3. Alex Liaugminas
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  4. Narasimham L. Parinandi
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  5. Periannan Kuppusamy
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Correspondence to Periannan Kuppusamy.

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Pandian, R.P., Kutala, V.K., Liaugminas, A. et al. Lipopolysaccharide-induced alterations in oxygen consumption and radical generation in endothelial cells. Mol Cell Biochem 278, 119–127 (2005). https://doi.org/10.1007/s11010-005-6936-x

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  • DOI: https://doi.org/10.1007/s11010-005-6936-x

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