Cell culture models for oxidative stress: superoxide and hydrogen peroxide versus normobaric hyperoxia

doi:10.1016/0921-8734(92)90043-O

Mutation Research/DNAging

Volume 275, Issues 3–6, September 1992, Pages 405-414

https://doi.org/10.1016/0921-8734(92)90043-O Get rights and content

Abstract

According to the free radical theory of aging, loss of cellular function during aging is a consequence of accumulating subcellular damage inflicted by activated oxygen species. In cells, the deleterious effects of activated oxygen species may become manifest when the balance between radical formation and destruction (removal) is disturbed creating a situation denoted as ‘oxidative stress’. Cell culture systems are especially useful to study the effects of oxidative stress, in terms of both toxicity and cellular adaptive responses. A better understanding of such processes may be pertinent to fully comprehend the cellular aging process.

This article reviews three model systems for oxidative stress: extracellular sources of O₂⁻ and H₂O₂, and normobaric hyperoxia (elevated ambient oxygen). Methodological and practical aspects of these exposure models are discussed, as well as their prominent effects as observed in cultures of Chinese hamster cell lines. Since chronic exposure models are to be preferred, it is argued that normobaric hyperoxia is a particularly relevant oxidative stress model for in vitro cellular aging studies.

References (80)

B.N. Ames
Endogenous DNA damage as related to cancer and aging
Mutation Res.
(1989)
U. Andrae et al.
Pyruvate and related alpha-ketoacids protect mammalian cells in culture against hydrogen-peroxide induced cytotoxicity
Toxicol. Lett.
(1985)
K.C. Bhuyan et al.
Regulation of hydrogen peroxide in eye tumors. Effects of 3-amino-1-H-1,2,4-triazole on catalase and glutathione peroxidase of rabbit eye
Biochim. Biophys. Acta
(1977)
M.O. Bradley et al.
Comparison of the effects of H₂O₂ and X-ray irradiation on toxicity, mutation and DNA damage/repair in mammalian cells (V79)
Biochim. Biophys. Acta
(1981)
O. Cantoni et al.
Hydrogen peroxide insult in cultured mammalian cells: relationships between DNA single-strand breakage, poly(ADP-ribose) metabolism and cell killing
Biochim. Biophys. Acta
(1989)
M.L. Cunningham et al.
Superoxide anion generated by potassium superoxide is cytotoxic and mutagenic to Chinese hamster ovary cells
Mutation Res.
(1983)
M.L. Cunningham et al.
Single-strand DNA breaks in rodent and human cells produced by superoxide anion or its reduction products
Mutation Res.
(1987)
M. Fiorani et al.
Inhibition of Chinese hamster ovary cell DNA synthesis by hydrogen peroxide
Chem.-Biol. Interact.
(1990)
H.J. Forman et al.
Superoxide radical and hydrogen peroxide in mitochondria
H.J. Forman et al.
Role of superoxide radical in mitochondrial dehydrogenase reactions
Biochem. Biophys. Res. Commun.
(1974)

S.M. Galloway et al.

Vitamin C is positive in the DNA synthesis inhibition and sister-chromatid exchanges tests

Mutation Res.

(1979)

J.J.P. Gille et al.

Effects of lethal exposure to hyperoxia and to hydrogen peroxide on NAD(H) and ATP pools in Chinese hamster ovary cells

Mutation Res.

(1989)

J.J.P. Gille et al.

Chromosomal instability in an oxygen-tolerant variant of Chinese hamster ovary cells

Mutation Res.

(1989)

J.J.P. Gille et al.

Effect of iron chelators on the cytotoxic and genotoxic action of hyperoxia in Chinese hamster ovary cells

Mutation Res.

(1992)

A.H. Hall et al.

Acute effects of a superoxide radical-generating system on DNA double-strand stability in Chinese hamster ovary cells. Determinatin by a modified fluorometric procedure

Mutation Res.

(1988)

L. Hayflick et al.

The serial cultivation of human diploid cell strains

Exp. Cell Res.

(1961)

B. Hennig et al.

Lipid peroxidation and endothelial cell injury: implications in atherosclerosis

Free Radical Biol. Med.

(1988)

M.E. Hoffmann et al.

Correlation between cytotoxic effects of hydrogen peroxide and the yield of DNA strand breaks in cells of different species

Biochim. Biophys. Acta

(1984)

P.A. Hyslop et al.

Mechanisms of oxidant-mediated cell injury. The glycolytic and mitochondrial pathways of ADP phosphorylation are major intracellular targets inactivated by hydrogen peroxide

J. Biol. Chem.

(1988)

D. Jamieson

Oxygen toxicity and reactive oxygen metabolites in mammals

Free Radical Biol. Med.

(1989)

H. Kappus

Lipid peroxidation: mechanisms, analysis, enzymology and biological relevance

J. Krall et al.

Superoxide mediates the toxicity of paraquat for cultured mammalian cells

J. Biol. Chem.

(1988)

M. Larramendy et al.

Iron-mediated induction of sister-chromatid exchanges by hydrogen peroxide and superoxide anion

Mutation Res.

(1987)

W.D. MacRae et al.

Induction of sister chromatid exchanges in Chinese hamster cells by reducing agents bisulfite and ascorbic acid

Toxicology

(1979)

R.J. Mehlhorn et al.

The free radical theory of aging: a critical review

Adv. Free Radical Biol. Med.

(1985)

A.C. Mello-Filho et al.

In vivo formation of single-strand breaks in DNA by hydrogen peroxide is mediated by the Haber-Weiss reaction

Biochim. Biophys. Acta

(1984)

H.P. Misra

Reaction of copper-zinc superoxide dismutase with diethyldithiocarbamate

J. Biol. Chem.

(1979)

H. Morrison et al.

Induction of DNA damage by menadione (2-methyl-1,4-naphthoquinone) in primary cultures of rat hepatocytes

Biochem. Pharmacol.

(1984)

Y. Oya et al.

The biological activity of hydrogen peroxide. IV. Enhancement of its clastogenic action by coadministration of histidine

Mutation Res.

(1988)

B.J. Phillips et al.

Genetic damage in CHO cells exposed to enzymically generated active oxygen species

Mutation Res.

(1984)

E. Pick et al.

A simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture

J. Immunol. Methods

(1980)

M. Sawada et al.

Induction of chromosomal aberrations in active oxygen-generating systems. II. A study with hydrogen peroxide-resistant cells in culture

Mutation Res.

(1988)

W.G.E.J. Schoonen et al.

Respiratory failure and stimulation of glycolysis in Chinese hamster ovary cells exposed to normobaric hyperoxia

J. Biol. Chem.

(1990)

H. Sies

Oxidative stress: introductory remarks

R.S. Sohal et al.

Effect of ambient oxygen concentration on lipofuscin accumulation in cultured rat myocytes — A novel in vitro model for lipofuscinogenesis

Free Radical Biol. Med.

(1989)

G. Speit

The relationship between the induction of SCEs and mutations in Chinese hamster cells. I. Experiments with hydrogen peroxide and caffeine

Mutation Res.

(1986)

D.R. Spitz et al.

Oxygen toxicity in control and H₂O₂-resistant Chinese hamster fibroblast cell lines

Arch. Biochem. Biophys.

(1990)

P. Tachon et al.

Histidine modulates the clastogenic effect of oxidative stress

Mutation Res.

(1989)

J.F. Turrens et al.

Ubisemiquinone is the electron donor for superoxide formation by complex III of heart mitochondria

Arch. Biochem. Biophys.

(1985)

K. Ziegler-Skylakakis et al.

Mutagenicity of hydrogen peroxide in V79 Chinese hamster cells

Mutation Res.

(1987)

Cited by (181)

Properties of an amorphous crystalline nanopowder Si–SiO<inf>2</inf> produced by pulsed electron beam evaporation
2024, Materials Chemistry and Physics
A mesoporous amorphous crystalline nanopowder (NP) Si–SiO₂ with a specific surface area (SSA) of 35 m²/g was produced by pulsed electron beam evaporation (PEBE) of a compact of micron commercial SiO₂ powder (0.9 m²/g). Si semimetal nanoparticles (NPles) reduced in for vacuum (≈4 Pa) had a marked effect on the thermal and luminescent properties of the Si–SiO₂ nanoparticles compared to the properties of amorphous SiO₂ NPles produced earlier by PEBE evaporation of a target compound made from NP Aerosil 90 (Degussa). Photocatalytic capabilities of Si–SiO₂ nanoparticles were evaluated. NPles Si–SiO₂ showed high antioxidant activity on HELA cancer cells.
Type 1 diabetes risk genes mediate pancreatic beta cell survival in response to proinflammatory cytokines
2022, Cell Genomics
We combined functional genomics and human genetics to investigate processes that affect type 1 diabetes (T1D) risk by mediating beta cell survival in response to proinflammatory cytokines. We mapped 38,931 cytokine-responsive candidate cis-regulatory elements (cCREs) in beta cells using ATAC-seq and snATAC-seq and linked them to target genes using co-accessibility and HiChIP. Using a genome-wide CRISPR screen in EndoC-βH1 cells, we identified 867 genes affecting cytokine-induced survival, and genes promoting survival and up-regulated in cytokines were enriched at T1D risk loci. Using SNP-SELEX, we identified 2,229 variants in cytokine-responsive cCREs altering transcription factor (TF) binding, and variants altering binding of TFs regulating stress, inflammation, and apoptosis were enriched for T1D risk. At the 16p13 locus, a fine-mapped T1D variant altering TF binding in a cytokine-induced cCRE interacted with SOCS1, which promoted survival in cytokine exposure. Our findings reveal processes and genes acting in beta cells during inflammation that modulate T1D risk.
Dependence of mitochondrial function on the filamentous actin cytoskeleton in cultured mesenchymal stem cells treated with cytochalasin B
2021, Journal of Bioscience and Bioengineering
Owing to their self-renewal and multi-lineage differentiation capability, mesenchymal stem cells (MSCs) hold enormous potential in regenerative medicine. A prerequisite for a successful MSC therapy is the rigorous investigation of their function after in vitro cultivation. Damages introduced to mitochondria during cultivation adversely affect MSCs function and can determine their fate. While it has been shown that microtubules and vimentin intermediate filaments are important for mitochondrial dynamics and active mitochondrial transport within the cytoplasm of MSCs, the role of filamentous actin in this process has not been fully understood yet. To gain a deeper understanding of the interdependence between mitochondrial function and the cytoskeleton, we applied cytochalasin B to disturb the filamentous actin-based cytoskeleton of MSCs. In this study we combined conventional functional assays with a state-of-the-art oxygen sensor-integrated microfluidic device to investigate mitochondrial function. We demonstrated that cytochalasin B treatment at a dose of 16 μM led to a decrease in cell viability with high mitochondrial membrane potential, increased oxygen consumption rate, disturbed fusion and fission balance, nuclear extrusion and perinuclear accumulation of mitochondria. Treatment of MSCs for 48 h ultimately led to nuclear fragmentation, and activation of the intrinsic pathway of apoptotic cell death. Importantly, we could show that mitochondrial function of MSCs can efficiently recover from the damage to the filamentous actin-based cytoskeleton over a period of 24 h. As a result of our study, a causative connection between the filamentous actin-based cytoskeleton and mitochondrial dynamics was demonstrated.
The modified Q-cycle: A look back at its development and forward to a functional model
2021, Biochimica et Biophysica Acta - Bioenergetics
Citation Excerpt :
The mechanistic basis for sensitivity to hyperoxia in isp-1(qm150) animals was unclear. Cell culture studies had shown that exposure to 100% O2 can mimic physiological oxidative stress and reactive oxygen species (ROS) generation [227,228], and isp-1(qm150) animals have been reported to produce a higher level of superoxide (SO) than wild type animals [229]. Thus, one possibility is that both hyperoxia and mutation of isp-1(qm150) increase ROS levels independently so as to exceed a threshold consistent with continued development.
On looking back at a lifetime of research, it is interesting to see, in the light of current progress, how things came to be, and to speculate on how things might be. I am delighted in the context of the Mitchell prize to have that excuse to present this necessarily personal view of developments in areas of my interests. I have focused on the Q-cycle and a few examples showing wider ramifications, since that had been the main interest of the lab in the 20 years since structures became available, - a watershed event in determining our molecular perspective. I have reviewed the evidence for our model for the mechanism of the first electron transfer of the bifurcated reaction at the Q_o-site, which I think is compelling. In reviewing progress in understanding the second electron transfer, I have revisited some controversies to justify important conclusions which appear, from the literature, not to have been taken seriously. I hope this does not come over as nitpicking. The conclusions are important to the final section in which I develop an internally consistent mechanism for turnovers of the complex leading to a state similar to that observed in recent rapid-mix/freeze-quench experiments, reported three years ago. The final model is necessarily speculative but is open to test.
Mitochondrial protective effects of PARP-inhibition in hypertension-induced myocardial remodeling and in stressed cardiomyocytes
2021, Life Sciences
During oxidative stress mitochondria become the main source of endogenous reactive oxygen species (ROS) production. In the present study, we aimed to clarify the effects of pharmacological PARP-1 inhibition on mitochondrial function and quality control processes.
L-2286, a quinazoline-derivative PARP inhibitor, protects against cardiovascular remodeling and heart failure by favorable modulation of signaling routes. We examined the effects of PARP-1 inhibition on mitochondrial quality control processes and function in vivo and in vitro. Spontaneously hypertensive rats (SHRs) were treated with L-2286 or placebo. In the in vitro model, 150 μM H₂O₂ stress was applied on neonatal rat cardiomyocytes (NRCM).
PARP-inhibition prevented the development of left ventricular hypertrophy in SHRs. The interfibrillar mitochondrial network were less fragmented, the average mitochondrial size was bigger and showed higher cristae density compared to untreated SHRs. Dynamin related protein 1 (Drp1) translocation and therefore the fission of mitochondria was inhibited by L-2286 treatment. Moreover, L-2286 treatment increased the amount of fusion proteins (Opa1, Mfn2), thus preserving structural stability. PARP-inhibition also preserved the mitochondrial genome integrity. In addition, the mitochondrial biogenesis was also enhanced due to L-2286 treatment, leading to an overall increase in the ATP production and improvement in survival of stressed cells.
Our results suggest that the modulation of mitochondrial dynamics and biogenesis can be a promising therapeutical target in hypertension-induced myocardial remodeling and heart failure.
HS-GC–MS analysis of volatile organic compounds after hyperoxia-induced oxidative stress: a validation study
2024, Intensive Care Medicine Experimental

View all citing articles on Scopus

View full text

Cell culture models for oxidative stress: superoxide and hydrogen peroxide versus normobaric hyperoxia

Abstract

Mutation Res.

Toxicol. Lett.

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Mutation Res.

Mutation Res.

Chem.-Biol. Interact.

Biochem. Biophys. Res. Commun.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Exp. Cell Res.

Free Radical Biol. Med.

Biochim. Biophys. Acta

J. Biol. Chem.

Free Radical Biol. Med.

J. Biol. Chem.

Mutation Res.

Toxicology

Adv. Free Radical Biol. Med.

Biochim. Biophys. Acta

J. Biol. Chem.

Biochem. Pharmacol.

Mutation Res.

Mutation Res.

J. Immunol. Methods

Mutation Res.

J. Biol. Chem.

Free Radical Biol. Med.

Mutation Res.

Arch. Biochem. Biophys.

Mutation Res.

Arch. Biochem. Biophys.

Mutation Res.