Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro

Introduction. Results on mitochondrial dysfunction in sepsis are controversial. We aimed to assess effects of LPS at wide dose and time ranges on hepatocytes and isolated skeletal muscle mitochondria. Methods. Human hepatocellular carcinoma cells (HepG2) were exposed to placebo or LPS (0.1, 1, and 1...

Full description

Bibliographic Details
Main Authors: Victor Jeger, Sebastian Brandt, Francesca Porta, Stephan M. Jakob, Jukka Takala, Siamak Djafarzadeh
Format: Article
Language:English
Published: Hindawi Limited 2015-01-01
Series:BioMed Research International
Online Access:http://dx.doi.org/10.1155/2015/353074
id doaj-f888f4e137ab49059e0b0ea4b1a389c4
record_format Article
spelling doaj-f888f4e137ab49059e0b0ea4b1a389c42020-11-24T21:05:35ZengHindawi LimitedBioMed Research International2314-61332314-61412015-01-01201510.1155/2015/353074353074Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In VitroVictor Jeger0Sebastian Brandt1Francesca Porta2Stephan M. Jakob3Jukka Takala4Siamak Djafarzadeh5Department of Intensive Care Medicine, Inselspital, Bern University Hospital and University of Bern, Freiburgstraße 10, 3010 Bern, SwitzerlandDepartment of Anesthesiology and Pain Therapy, Inselspital, Bern University Hospital and University of Bern, 3010 Bern, SwitzerlandDepartment of Intensive Care Medicine, Inselspital, Bern University Hospital and University of Bern, Freiburgstraße 10, 3010 Bern, SwitzerlandDepartment of Intensive Care Medicine, Inselspital, Bern University Hospital and University of Bern, Freiburgstraße 10, 3010 Bern, SwitzerlandDepartment of Intensive Care Medicine, Inselspital, Bern University Hospital and University of Bern, Freiburgstraße 10, 3010 Bern, SwitzerlandDepartment of Intensive Care Medicine, Inselspital, Bern University Hospital and University of Bern, Freiburgstraße 10, 3010 Bern, SwitzerlandIntroduction. Results on mitochondrial dysfunction in sepsis are controversial. We aimed to assess effects of LPS at wide dose and time ranges on hepatocytes and isolated skeletal muscle mitochondria. Methods. Human hepatocellular carcinoma cells (HepG2) were exposed to placebo or LPS (0.1, 1, and 10 μg/mL) for 4, 8, 16, and 24 hours and primary human hepatocytes to 1 μg/mL LPS or placebo (4, 8, and 16 hours). Mitochondria from porcine skeletal muscle samples were exposed to increasing doses of LPS (0.1–100 μg/mg) for 2 and 4 hours. Respiration rates of intact and permeabilized cells and isolated mitochondria were measured by high-resolution respirometry. Results. In HepG2 cells, LPS reduced mitochondrial membrane potential and cellular ATP content but did not modify basal respiration. Stimulated complex II respiration was reduced time-dependently using 1 μg/mL LPS. In primary human hepatocytes, stimulated mitochondrial complex II respiration was reduced time-dependently using 1 μg/mL LPS. In isolated porcine skeletal muscle mitochondria, stimulated respiration decreased at high doses (50 and 100 μg/mL LPS). Conclusion. LPS reduced cellular ATP content of HepG2 cells, most likely as a result of the induced decrease in membrane potential. LPS decreased cellular and isolated mitochondrial respiration in a time-dependent, dose-dependent and complex-dependent manner.http://dx.doi.org/10.1155/2015/353074
collection DOAJ
language English
format Article
sources DOAJ
author Victor Jeger
Sebastian Brandt
Francesca Porta
Stephan M. Jakob
Jukka Takala
Siamak Djafarzadeh
spellingShingle Victor Jeger
Sebastian Brandt
Francesca Porta
Stephan M. Jakob
Jukka Takala
Siamak Djafarzadeh
Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro
BioMed Research International
author_facet Victor Jeger
Sebastian Brandt
Francesca Porta
Stephan M. Jakob
Jukka Takala
Siamak Djafarzadeh
author_sort Victor Jeger
title Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro
title_short Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro
title_full Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro
title_fullStr Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro
title_full_unstemmed Dose Response of Endotoxin on Hepatocyte and Muscle Mitochondrial Respiration In Vitro
title_sort dose response of endotoxin on hepatocyte and muscle mitochondrial respiration in vitro
publisher Hindawi Limited
series BioMed Research International
issn 2314-6133
2314-6141
publishDate 2015-01-01
description Introduction. Results on mitochondrial dysfunction in sepsis are controversial. We aimed to assess effects of LPS at wide dose and time ranges on hepatocytes and isolated skeletal muscle mitochondria. Methods. Human hepatocellular carcinoma cells (HepG2) were exposed to placebo or LPS (0.1, 1, and 10 μg/mL) for 4, 8, 16, and 24 hours and primary human hepatocytes to 1 μg/mL LPS or placebo (4, 8, and 16 hours). Mitochondria from porcine skeletal muscle samples were exposed to increasing doses of LPS (0.1–100 μg/mg) for 2 and 4 hours. Respiration rates of intact and permeabilized cells and isolated mitochondria were measured by high-resolution respirometry. Results. In HepG2 cells, LPS reduced mitochondrial membrane potential and cellular ATP content but did not modify basal respiration. Stimulated complex II respiration was reduced time-dependently using 1 μg/mL LPS. In primary human hepatocytes, stimulated mitochondrial complex II respiration was reduced time-dependently using 1 μg/mL LPS. In isolated porcine skeletal muscle mitochondria, stimulated respiration decreased at high doses (50 and 100 μg/mL LPS). Conclusion. LPS reduced cellular ATP content of HepG2 cells, most likely as a result of the induced decrease in membrane potential. LPS decreased cellular and isolated mitochondrial respiration in a time-dependent, dose-dependent and complex-dependent manner.
url http://dx.doi.org/10.1155/2015/353074
work_keys_str_mv AT victorjeger doseresponseofendotoxinonhepatocyteandmusclemitochondrialrespirationinvitro
AT sebastianbrandt doseresponseofendotoxinonhepatocyteandmusclemitochondrialrespirationinvitro
AT francescaporta doseresponseofendotoxinonhepatocyteandmusclemitochondrialrespirationinvitro
AT stephanmjakob doseresponseofendotoxinonhepatocyteandmusclemitochondrialrespirationinvitro
AT jukkatakala doseresponseofendotoxinonhepatocyteandmusclemitochondrialrespirationinvitro
AT siamakdjafarzadeh doseresponseofendotoxinonhepatocyteandmusclemitochondrialrespirationinvitro
_version_ 1716768241283497984