ISSN 0096-3925, Moscow University Biological Sciences Bulletin, 2018, Vol. 73, No. 4, pp. 199–202.

© Allerton Press, Inc., 2018.

Original Russian Text © M.A. Chelombitko, 2018, published in Vestnik Moskovskogo Universiteta, Seriya 16: Biologiya, 2018, Vol. 73, No. 4, pp. 242–246.

REVIEWS

Role of Reactive Oxygen Species in Inflammation:

A Minireview
M. A. Chelombitko
Department of Biology, Moscow State University, Moscow, 119234 Russia
e-mail: chelombitko@mail.bio.msu.ru
Received May 29, 2018; in final form, September 5, 2018

Abstract—Inflammation is a protective response of a multicellular organism to injury in order to localize,

eliminate, and remove harmful stimuli as well as to recover (or replace) damaged tissues. There recently has
been increasing evidence that reactive oxygen species (ROS) are involved in the initiation, progression, and
resolution of the inflammatory response. Furthermore, ROS act as microbicidal agents and second messen-
gers in the intracellular signaling. The latter function is performed via posttranslational modification of pro-
tein-associated redox-sensitive cysteine residues that can undergo oxidation. At the same time, there is clear
evidence that overproduction of ROS may result in cell and tissue injury and contribute to chronic inflam-
mation underlying many neurodegenerative, cardiovascular, and metabolic diseases. This review has focused
on the role of ROS in the key inflammatory events (increased vascular permeability and leukocyte extravasa-
tion, respiratory burst and phagocytosis, and angiogenesis) and some events leading to the resolution of
inflammation. In addition, the pathological function of ROS in oxidative stress is discussed.

Keywords: reactive oxygen species, inflammation, oxidative stress, NADPH-oxidase, respiratory burst, mito-
chondria.
DOI: 10.3103/S009639251804003X

Reactive oxygen species (ROS) are free radicals
(molecules or molecular fragments with one or more
unpaired electrons in atomic or molecular orbitals)
derived from molecular oxygen. ROS include super-
oxide anion (O2− ), hydroxyl (HO•), and hydroperoxyl
(HOi2 ) radicals; hydrogen peroxide (H2O2), nitric
oxide (NO), hypochlorous acid (HOCl), and singlet
oxygen (1O2) [1].
The main source of ROS in many cell types is mito-
chondria where ROS are generated by the electron
transport chain (from complexes of the mitochondrial
respiratory chain: complex I—NADH dehydrogenase
complex; complex III—cytochrome bc1complex) as
well as dehydrogenases in the mitochondrial matrix,
p66shc protein in the intermembrane space, and
monoamine oxidase in the outer membrane. ROS can
be also produced by other enzymes in the cell, such as
xanthine oxidase, cyclooxygenases, myeloperoxidase,
NADPH oxidase, cytochrome P450 enzymes, lipoxy-
genases, and oxidoreductase 1α [2].
High levels of ROS incur damage to DNA, lipids,
and proteins. However, at low concentrations, ROS
act as important mediators involved in the regulation
of cell growth, cell adhesion, differentiation, cell
death, etc. ROS are involved in intracellular signaling
through posttranslational modification of redox-sen-
sitive proteins, including various receptors and chan-
nels, kinases and phosphatases, caspases, and tran-
scriptional factors. Redox-sensitive proteins comprise
proteins that contain functionally significant residues
of cysteine that can undergo oxidation. Thus, H2O2
can oxidize sulfhydryl groups of cysteine (Cys–SH)
with the formation of sulfenic (Cys–SOH) and then
sulfinic (Cys–SO2H) and sulfonic (Cys–SO3H) acids.
These modifications alter the protein activity and
affect the protein functioning in the pathway of signal
transduction [3].
Considerable data have now been accumulated that
indicate that ROS participate in the initiation, pro-
gression, and resolution of inflammation. We further
discuss the involvement of ROS in the critical events
of this process.
Vascular Permeability and Leukocyte Extravasation
In the initial stage of an inflammatory process, the
cells of innate immunity (mainly macrophages and
mast cells) activated by injurious agents produce vari-
ous inflammatory mediators leading to increased vas-
cular permeability and leukocyte migration towards
the site of injury [4].
Numerous data confirm the important role of ROS
in the regulation of this stage of inflammation. Thus,

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ROS regulate expression of the intercellular adhesion
molecules (such as ICAM-1, VCAM-1, P- and
E-selectin) on the endothelial surface that ensures its
interaction with blood leukocytes and further transen-
dothelial migration of leukocytes [5]. Overexpression
of the antioxidant enzyme superoxide dismutase
decreases leukocyte binding to endothelium at the site
of inflammation by reducing the expression of adhe-
sion molecules [6].
A gradient of H2O2 generated at the site of injury is
one of the early factors of leukocyte recruitment [7].
Such cytokines as tumor necrosis factor (TNF),
angiopoietin-1, platelet-derived growth factor
(PDGF), vascular endothelial growth factor (VEGF),
and a number of components of the extracellular
matrix stimulate cell migration and adhesion, which is
accompanied by activation of NADPH oxidase [8].
Neutrophils with a deficient NADPH oxidase activ-
ity exhibit impaired migration to the site of injury in
mice [9].
Furthermore, ROS act upon the endothelial per-
meability by affecting cell–cell contacts. Thus, occlu-
dins, proteins of tight junctions, are redox sensitive,
and oxidative stress reduces the occludin expression.
We also know that occludin oligomerization is affected
by the antioxidant function of glutathione in the cell.
Under conditions of inflammation, the balance
between reduced and oxidized glutathione shifts in
favor of the oxidized form, which results in progres-
sion of oxidative stress and the impaired assembly of
occludins that increases the permeability of tight junc-
tions [10].
Moreover, ROS affect another type of inter-endo-
thelial junctions—adherens junctions. This is mainly
mediated by phosphorylation of key serine and tyro-
sine residues of the relevant proteins (VE-cadherin,
β-catenin, and p120 catenin), which induces junc-
tional disassembly [5]. An important role in the dis-
ruption of adherens junctions is played by a small
GTPase Rac1 from the RhoGTPase family. Thus, it
has been shown that treatment of human pulmonary
microvascular endothelial cells (HMVECs) with
cytokine VEGF induces the Rac1-mediatiated gen-
eration of ROS that leads to phosphorylation of
VE-cadherin and β-catenin and loss of adherens
junction integrity [11].
Respiratory Burst and Phagocytosis
The most well-known role of ROS in inflamma-
tion is their participation in the phenomenon of
″respiratory burst.″ Once neutrophils and macro-
phages have arrived at the site of inflammation, they
phagocytize pathogens and cellular debris. This pro-
cess induces the NADPH oxidase assembly and acti-
vation at the phagosome membrane. This enzyme
reduces O2 to O2− that triggers other ROS. Respiratory
burst-derived ROS are released not only into phago-
MOSCOW UNIVERSITY BIOLOGICAL SCIENCES BULLETIN
somes but also into the surrounding tissues. They
mainly act as bactericidal agents [12]. It should be
noted that, depending on the pathogen size, NADPH
oxidase is assembled either at the plasma membrane or
at the membrane of neutrophil phagosomes. Thus,
microbes of small size trigger ROS intracellularly, sup-
pressing the interleukin IL-1β expression and limiting
the recruitment of new neutrophils. Large pathogens
trigger ROS extracellularly, amplifying the IL-1β
expression that leads to the formation of neutrophil
clusters around pathogens [13]. NADPH oxidase plays
an important role in the progression of the inflamma-
tory response not only because of its involvement in
the respiratory burst. This enzyme is also required for
the formation of neutrophil extracellular traps [14].
Angiogenesis
Angiogenesis is a process of the growth of new ves-
sels and is an important stage in the proliferative phase
of inflammation. There are evidences of the key role of
NADPH oxidase-derived ROS in this phase of the
inflammatory response. It has been shown in human
umbilical vein endothelial cells (HUVECs) that
NADPH oxidase is localized at the leading edge of
migrating cells and is linked to actin and IQGAP1 pro-
tein (Ras GTPase-activating-like protein). The dis-
ruption of this binding leads to the NADPH oxidase
translocation from the leading edge and the impaired
migration of endothelial cells [15]. The effect of ROS
on cell migration is apparently mainly mediated
through the redox sensitivity of small Rho GTPases
involved in cytoskeleton rearrangements. Thus, in the
HUVECs, VEGF induced migration of endothelial
cells, accompanied by increased production of ROS in
mitochondria. The targeted mitochondria delivery of
the vitamin E-based antioxidant (Mito-Vit-E), as well
as overexpression of mitochondrial catalase, inhibited
VEGF-mediated production of mitochondrial ROS,
activation of GTPase Rac1, and cell migration. The
use of Mito-Vit-E did not affect cell migration in the
experiments with constitutively active GTPase Rac1.
These results provide strong evidence that mitochon-
dria-derived ROS regulate cell migration via Rac1
[16].
Resolution of Inflammation
There is increasing evidence of ROS involvement
in a number of processes that contribute to the natural
resolution of inflammation. One of the key events pre-
ceding inflammation resolution is massive death of
neutrophils. Signaling pathways regulating the apop-
tosis of granulocytes, and other cells are ROS-depen-
dent. Thus, in a model of antigen-induced arthritis in
mice, H2O2 has been demonstrated to induce the neu-
trophil apoptosis and resolution of the inflammatory
response [17].
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 ROLE OF REACTIVE OXYGEN SPECIES IN INFLAMMATION
Another important event contributing to transition
to the resolution phase of inflammation is phagocyto-
sis of apoptotic cells by M2-polarized macrophages
exhibiting anti-inflammatory activity. There is evi-
dence that the interaction of macrophages with apop-
totic cells leads to destabilization of the NADPH oxi-
dase mRNA by the RNA-binding protein SYNCRIP
(Synaptotagmin binding, cytoplasmic RNA interact-
ing protein). This consequently decreases ROS pro-
duction and activates macrophages toward an M2-like
phenotype [18]. At the same time, the formation of
ROS plays an important role in the differentiation of M2
macrophages. Thus, inhibition of the NADPH-
dependent production of O2− by butylated hydroxy-
anisole specifically blocks the differentiation of M2
macrophages without affecting the differentiation of
proinflammatory M1-macrophages [19]. It is worth
noting that the microbicidal function of macrophages
M1 requires continuous production of ROS by the
inducible NO synthase, NADPH oxidase, and by
mitochondria. ROS play an important role in main-
taining the proinflammatory phenotype of macro-
phages also due to the activation of the NF-κB signal-
ing pathway (Nuclear factor kappa-light-chain-
enhancer of activated B cells) involved in the expres-
sion of proinflammatory cytokines [20].
It is known that ROS have both activating and
inhibiting roles in NF-κB signaling. ROS activate the
transcription factor NF-κB mainly through inactivat-
ing phosphorylation of its inhibitory protein IkBα.
Thus, H2O2 affects the phosphorylation of IkBα on
tyrosine residues, which leads to IkBα degradation
and NF-κB activation. At the same time, the direct
action of ROS on the NF-κB heterodimer inhibits its
DNA binding ability. In turn, the NF-κB signaling
pathway may affect intracellular levels of ROS via
increased expression of various antioxidant proteins
(superoxide dismutase, catalase, thioredoxin-1 and
thioredoxin-2, etc.). Furthermore, NF-κB also acti-
vates the transcription of a number of enzymes
involved in ROS production, such as NADPH oxi-
dase, inducible NO synthase, cyclooxygenase-2, and
5- and 12-lipoxygenases [21].
Oxidative Stress
Although ROS are important regulatory mole-
cules in almost all stages of the inflammatory pro-
cess, ROS overproduction by mitochondria and
NADPH oxidase of leukocytes and endothelial cells,
which is not compensated by the antioxidant sys-
tems, may result in serious cellular and tissue damage
and promote chronic inf lammation underlying many
neurodegenerative, cardiovascular, and metabolic
diseases [22].
Progression of oxidative stress in the cell results in
the disturbance of cellular redox status, which shifts
towards oxidation processes. At the same time, ROS
MOSCOW UNIVERSITY BIOLOGICAL SCIENCES BULLETIN
201
start to adversely affect different components of the
cell. Lipid peroxidation of ω-3 and ω-6 polyunsatu-
rated fatty acids of membrane phospholipids causes
damage to cell membranes. Furthermore, lipid perox-
idation generates such relatively stable end-products as
4-hydroxy-2-nonenal and malondialdehyde, which
may themselves exhibit cytotoxic effects. Oxidation of
tyrosine, serine, and cysteine residues in proteins
results in disturbance of protein functions. Progres-
sion of oxidative stress results in single and/or double-
stranded breaks in DNA. Mitochondrial damage
caused by oxidative stress is followed by a decrease in
the transmembrane potential, changes in the mem-
brane permeability, and accelerated release of apop-
totic factors. All of the above factors impair cell func-
tionality, which leads to cell death [3, 23].
Therefore, the currently available information
about the ROS-induced effects on inflammation indi-
cate their multifaceted but still understudied role in
this process. At the same time, ROS are known not
only as microbicidal agents but they are also involved
in intracellular signaling transduction, and their over-
production may contribute to chronic inflammation.
Considerable data have now been accumulated
demonstrating the success of antioxidant therapy in
the complex treatment of inflammatory diseases [24].
Moreover, in recent years, the number of research and
development studies related to the anti-inflammatory
properties of mitochondria-targeted antioxidants has
increased. The use of such compounds in the treat-
ment of inflammatory diseases is highly promising,
since mitochondria are one of the main sources of
ROS generation contributing to increased oxidative
stress and playing a key role in the pathogenesis of
inflammatory diseases [1, 25]. Studying the role of
ROS that originated from different sources in the
inflammatory response regulation is important for
gaining a better understanding of the ROS involve-
ment in biological processes as well as for providing
insights into the mechanisms of protective action of
therapeutic agents with antioxidant properties, which
would eventually form the basis for the development of
more effective anti-inflammatory drugs.
ACKNOWLEDGMENTS
The work was financially supported by the Russian
Science Foundation (project no. 14-50-00029).
COMPLIANCE
WITH ETHICAL STANDARDS
The authors declare that they have no conflict of
interest. This article does not contain any studies
involving animals or human participants performed by
any of the authors.
Vol. 73 No. 4 2018
202 CHELOMBITKO
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Translated by M. Romanova
Vol. 73 No. 4 2018