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Journal of Sport and Health Science 2 (2013) 67e74

www.jshs.org.cn

Review

Mitochondrial redox metabolism in aging: Effect of exercise interventions

Hai Bo a,b, Ning Jiang a, Li Li Ji a,c, Yong Zhang a,*
a

Tianjin Key Laboratory of Exercise Physiology and Sports Medicine, Department of Health & Exercise Science, Tianjin University of Sport,
Tianjin 300381, China
b
Department of Military Training Medicines, Logistics University of Chinese Peoples Armed Police Force, Tianjin 300162, China
c
Laboratory of Physiological Hygiene and Exercise Science, School of Kinesiology, University of Minnesota, Minneapolis, MN 55455, USA
Received 12 December 2012; revised 20 January 2013; accepted 22 February 2013

Abstract
Mitochondrial redox metabolism has long been recognized as being central to the effects of aging and the development of age-related
pathologies in the major oxidative organs. Consistent evidence has shown that exercise is able to retard the onset and impede the progression of
aging by modifying mitochondrial oxidanteantioxidant homeostasis. Here we provide a broad overview of the research evidence showing the
relationship between mitochondrial redox metabolism, aging and exercise. We address part aspects of mitochondrial reactive oxygen species
(ROS) metabolism, from superoxide production to ROS detoxification, especially antioxidant enzymes and uncoupling protein. Furthermore, we
describe mitochondrial remodeling response to aging and exercise, which is accompanied by bioenergetics and redox regulation. In addition,
potential mechanisms for redox signaling involved in mitochondrial remodeling and redox metabolism regulation are also reviewed.
Copyright 2013, Shanghai University of Sport. Production and hosting by Elsevier B.V. All rights reserved.
Keywords: Aging; Exercise; Mitochondrial remodeling; PGC-1a; Reactive oxygen species

1. Introduction
The aging process results in a gradual and progressive
structural and functional deterioration of biomolecules that is
associated with many pathological conditions, including
cancer, neurodegenerative diseases, sarcopenia, and liver
dysfunction. In the 1950s, Harman1 first proposed the free
radical theory of aging. This theory contended that free
radicals, produced from normal metabolism, lead to

* Corresponding author.
E-mail address: yzhang@tjus.edu.cn (Y. Zhang)
Peer review under responsibility of Shanghai University of Sport

Production and hosting by Elsevier

irreversible cell damage and an overall functional decline

during aging. In the 1970s, he extended the theory to implicate
mitochondrial production of reactive oxygen species (ROS,
such as O2  and H2O2) as the main cause for age-related
macromolecular damage.2
Epidemiological studies clearly demonstrate that endurance exercise reduces the risk of chronic diseases and extends
life expectancy.3 Consistent evidence has shown that exercise
is able to retard the onset and impede the progression of
aging by modifying mitochondrial oxidanteantioxidant
homeostasis.4 However, there are still unanswered basic
scientific questions in the implication of exercise in modulating aging. Especially, how increased ROS production
during physical exercise could be involved in exerciseinduced health-promoting benefits. In this review, we
consider the evidence that specific aspects of mitochondrial
redox metabolism affect aging. We then discuss redoxsensitive signal transduction pathways associated with exercise in modulating aging.

2095-2546/$ - see front matter Copyright 2013, Shanghai University of Sport. Production and hosting by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.jshs.2013.03.006

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H. Bo et al.

2. Mitochondrial ROS production and aging

In the mitochondrial respiratory chain, under conditions in
which complexes I and III are highly reduced, a significant
proportion of electrons are diverted from them directly to
molecular oxygen, thus forming the radical superoxide (O2 ).
In addition, it recently has been shown that p66shc, a protein
present in the intermembrane space, can also catalyze O2 
production via electron transfer from reduced cytochrome c to
molecular oxygen.5 Although O2  is not membrane permeable, it can be spontaneously or catalytically dismutated to
hydrogen peroxide (H2O2), which can efficiently pass through
biomembranes and therefore acts not only at its site of
formation but also elsewhere.
At increased abundance, ROS are able to damage cellular
components such as proteins, nucleic acids, and lipids by the
abstraction of electrons. The damaging potential of mitochondrial ROS is the kernel of the mitochondrial free radical
theory of aging, which suggests that mitochondrial ROS lead
to an accumulation of molecular damage over time and, after
passing certain thresholds, to cellular degeneration and death.
In general, ROS production is found to be increased in cells
and is associated with aging. Furthermore, genetic studies in
worm, fly, and mouse have linked enhanced stress resistance
or reduced free radical production with increased lifespan.
Because mitochondria are the major producer of ROS in
mammalian cells, the close proximity to ROS places mitochondrial prone to oxidative damage. During aging, the
increase in ROS production is associated with increased levels
of 8-oxoguanine (8-oxoG) from mitochondrial DNA (mtDNA)
oxidation, increased nitration of complex II, and altered
carbonylation of complex I, complex V, and isocitrate dehydrogenase. Oxidative damage induced impaired function of
respiratory chain will in turn trigger further accumulation of
ROS, which results in a vicious cycle leading to energy
depletion and ultimately cell death.

3. Mitochondrial antioxidant enzymes in aging

To avoid the potentially damaging effects of ROS, cells use
a variety of systems to keep ROS within tolerable limits. This
is especially important in mitochondria because the enzymes
that drive ATP production are rich in thiol residues that can be
irreversibly oxidized. Superoxide generated by mitochondria
is quickly dismutated to H2O2 by either manganese (Mn) or
copper/zinc (Cu/Zn)-dependent superoxide dismutase (SOD),
which are located in the matrix and intermembrane space,
respectively. Although H2O2 is the main ROS used in
signaling, it is degraded by a variety of enzymes and low
molecular weight molecules to prevent oxidative stress. The
chief H2O2-quenching systems in mitochondria include the
GSH/glutathione peroxidase (GPx) system, peroxiredoxins,
thioredoxins, and glutaredoxins. Catalase (CAT) also degrades
H2O2 but only when H2O2 reaches high concentrations.
MnSOD is an intramitochondrial free radical scavenging
enzyme that is the first line of defense against O2  produced.
Over-expression of MnSOD in Drosophila melanogaster was

shown to have beneficial effects, extending lifespan.6

Furthermore, the tissue-specific over-expression of MnSOD
in motor neurons was sufficient to mediate the same effect.
Conversely, complete ablation of this enzyme in mice causes
severe oxidative damage to mitochondria and early postnatal
death associated with dilated cardiomyopathy and neurodegeneration.7 MnSOD activity, both in rats and men, are
reported to increase significantly with aging and to respond
to age-related oxidative stress in the mitochondria by upregulation. However, MnSOD protein and mRNA levels was
unchanged or decreased with age in most tissues. In addition,
binding of transcription factor nuclear factor-kB (NF-kB) and
transcription factor activator protein-1 (AP-1) to corresponding oligonucleotide sequences present in the MnSOD gene
were decreased in aged cells.8 Both NF-kB and AP-1 binding
sites are present in the promoter of the mammalian MnSOD
gene, and oxidative stress has been shown to activate their
binding. A decrease in the binding of these nuclear factors
despite increased ROS generation observed in aged cells
suggests that molecular signaling of MnSOD gene expression
is retarded due to aging. The observed increase in MnSOD
activity in some tissues appears to be due to a posttranslational
modification (activation) of the enzyme molecules during
aging.
CAT is widely distributed in the cell, with the majority of
activity occurring in the mitochondria and peroxisomes, where
it functions to catalyze the decomposition of H2O2 to water
and oxygen. Transgenic mice that overexpress mitochondrial
CAT have increased median and maximum lifespans, attenuated age-related changes including decline of diastolic function, myocardial performance as well as ventricular fibrosis.9
CAT largely determines the functional antioxidant capacity
of mitochondria and is the enzyme that is most affected in
aging.10 In rats and mice cellular CAT levels drop with age,
which is accompanied by an increase in oxidative stress.
Chronically reducing CAT activity causes cells to display
a cascade of accelerated aging reactions.11 Sirtuin1 (Sirt1) has
been shown to affect CAT expression and be a determinant of
cell apoptosis by regulating cellular ROS levels. NADdependent deacetylase Sirt1 maintains cell survival by regulating CAT expression and by preventing the depletion of ROS
required for cell survival. In contrast, excess ROS upregulates
Sirt1, which activates CAT leading to rescuing apoptosis.12
4. Exercise interventions affecting mitochondrial
antioxidant enzymes
There is an abundance of literature reporting muscle antioxidant adaptation to chronic exercise training and several
reviews have also addressed this topic thoroughly. Among
antioxidant enzymes in skeletal muscle, MnSOD activity has
consistently been shown to increase with exercise training in
an intensity-dependent manner. GPx activity has also shown to
increase after endurance training by most authors. GSH plays
a critical role in muscle antioxidant defense during exercise.
Several studies have demonstrated that GSH content and the
rate-limiting enzyme for GSH synthesis, a-glutamylcysteine

Mitochondrial redox metabolism in aging and exercise

synthetase (GCS), can be induced by endurance training in rat

skeletal muscle. However, training effect on CAT activity has
been inconsistent and controversial.13
As mentioned previously several oxidative stress-sensitive
signaling pathways are operational in mammalian systems and
play an important role in maintaining cellular oxidanteantioxidant balance. One of the most important involves
the NF-kB. Several antioxidant enzymes contain NF-kB
binding sites in their gene promoter region, such as MnSOD,
inducible nitric oxide synthetase, and GCS. Therefore, they
can be potential targets for exercise-activated upregulation via
the NF-kB signaling pathway. Ji et al.14 reported that an acute
bout of treadmill running activated NF-kB binding in rat
skeletal muscle 2 h after exercise. Furthermore, the NF-kB
signaling pathway was activated in a redox-sensitive manner
during muscular contraction.14 Consistently, Gomez-Cabrera
et al.15 have demonstrated that marathon running induces
activation of the p50 subunit of the NF-kB complex in
lymphocytes. This effect was prevented by treatment with
allopurinol.
It is well documented that a single episode of muscular
contraction can activate the MAPK pathway in human skeletal
muscle. As members of MAPK family, extracellular signalregulated kinase (ERK) and p38 MAPK activity can lead to
the sequential phosphorylation of a series of proteins, resulting
in increased expression AP-1, which is an important DNAbinding site on many genes responsive to oxidative stress.16 It
was also found that ROS generated during exercise activate
MAPKs, which in turn activate NF-kB, resulting in an
increased expression of important enzymes associated with
cell antioxidant defense (MnSOD and GPx). Prevention of
ROS formation by inhibition of XO abolishes these effects.17
This result showed the role of cross-talk in the MAPK and
NF-kB pathway in exercise-activated antioxidant signaling.
5. Uncoupling proteins, mitochondrial ROS production,
and aging
It is well established that there is a strong positive correlation between mitochondrial membrane potential (Dj) and
ROS production. It has been reported that, at high Dj, even
a small increase in membrane potential gives rise to a large
stimulation of H2O2 production. Similarly, only a small
decrease in Dj (10 mV), which is termed mild uncoupling,
is able to inhibit H2O2 production by 70%. Hence, the mild
uncoupling of mitochondrial oxidative phosphorylation may
represent the first line of defense against oxidative stress.
Brand18 first proposed the uncoupling to survive hypothesis,
which states that increased uncoupling leads to greater oxygen
consumption and reduces the proton motive force by reducing
Dj which subsequently reduces ROS generation. Further
evidence supports this hypothesis as a number of studies show
higher mitochondrial respiration and greater mitochondrial
uncoupling increases lifespan in a diverse range of organisms.
Uncoupling proteins (UCPs), a heterogeneous family of
proteins located in the mitochondrial inner membrane, appear
to be a logical candidate to achieve mild uncoupling. UCP2

69

and UCP3 are w73% homologous to each other and w58%

homologous to the thermogenic protein, UCP1. UCP3 is
highly selectively expressed in skeletal muscle and brown fat,
whereas UCP2 is ubiquitously expressed. It has been critically
examined that UCP2/UCP3 become true uncoupling proteins
when stimulated by ROS or 4-hydroxynonenal, and thus
become capable of diminishing oxidative damage by a feedback mechanism activating mild uncoupling.19 Evidence for
this includes observations that mice deficient in UCP3 have
higher levels of membrane lipid peroxidative damage.20
Numerous studies have demonstrated that UCP2 overexpression diminishes ROS production in a large number of
tissues examined, including the heart, pancreas, adipose tissue,
muscle, immune system, spleen, and steatotic liver.21
Andrews and Horvath22 reported that UCP2 gene deletion
dramatically reduces maximum lifespan in mice, and transgenic UCP2 overexpression can partially rescue the early
neonatal lethality of MnSOD deficiency. Similar observations
have been made with UCP3. Mitochondria from UCP2
knockout mice produce more ROS and contain more oxidative
damage in comparison to wild-type.23 In addition, overexpression of UCP3 has been found to lower the increased
ROS formation associated with aging.24 However, simultaneous overexpression of UCP2 and UCP3 has minimal effect
on longevity.19 This could be related to the requirement of
these proteins for effector molecules. Maximal activity of
UCPs requires stimulation with fatty acids, including end
products of fatty acid peroxidation. UCPs activities are also
inhibited by adenine nucleotides. Thus, increasing UCP
expression without modifying levels of stimulators and
inhibitors of their activities may not affect membrane potential
under normal conditions.
It has been repeatedly demonstrated that aging is associated
with an increase in oxidative stress, and therefore an increase
in uncoupling may be expected with aging. However, there
was a dramatic reduction of UCP3 content associated with
decreased state 4 respiration of skeletal muscle mitochondria
from old rats.25 In addition, it was found that muscle with the
most active fiber type (type I) and higher resting O2 uptake had
mild uncoupling in adults but stable mitochondrial function
into old age. This protection of mitochondrial function
suggests that mild uncoupling is a mechanism that reduces the
ROS production, ameliorates mitochondrial damage, and
assists fiber survival with age. In contrast, muscle with greater
content of the least active fiber type (type II) and lower resting
O2 uptake was well coupled in adults yet showed defect in
mitochondrial coupling and depletion of ATP with age
consistent with ROS damage. Thus, mild uncoupling may
explain the paradox of greater longevity in the most active
fibers by providing a protective mechanism that impacts the
pace of cellular aging.26
6. Mitochondrial uncoupling in response to exercise
Several studies have shown that UCP3 expression was
increased in response to an acute bout of exercise or
contractile activity in mammalian skeletal muscle.27 We

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H. Bo et al.

investigated UCP2 and UCP3 expression along with mitochondrial respiratory function and ROS generation in rat heart
and skeletal muscle during and after an acute bout of prolonged exercise up to 2.5 h. The data demonstrate that UCP2/
UCP3 expression can be rapidly upregulated during prolonged
exercise, and a coordinated regulation to reduce ROS generation and to preserve mitochondrial respiratory function.28,29
Noticeably, MnSOD expression and enzyme activity was not
up-regulated until after exercise under this experimental
condition. Thus, our data suggest that during an acute bout of
demanding exercise, the primary strategy of mitochondria to
reduce oxidative stress is decreasing the production of O2  by
overexpressing UCP2/UCP3, instead of enhancing the
removal of O2  by overexpressing MnSOD.
Our study also demonstrated that endurance training
attenuated acute stress-induced UCP2/UCP3 expression and
activity in rat. Meanwhile, endurance training significantly
elevated MnSOD activity and protein level.28,30 Similarly,
Noland et al.31 reported that acute exercise increases skeletal
muscle UCP3 expression in untrained but not trained humans.
These findings raised some interesting questions regarding the
relationship among mitochondrial respiratory uncoupling,
ROS generation, antioxidant defense, and ATP synthesis. An
ameliorated exercise response of UCP2/UCP3 after training
could be viewed as a compensatory mechanism to avoid
excessive sacrifice of oxidative phosphorylation efficiency and
heat production. McLeod et al.32 reported that in mitochondria
from SOD2 knockout mice, ATP production was markedly
reduced due to a higher O2  concentration which activated
UCP2 excessively. Taken together, exercise training adaptation
of MnSOD may enhance mitochondrial tolerance to ROS
production and hence a smaller UCP2/UCP3 activation during
intensive exercise, thus protecting the efficiency of oxidative
phosphorylation.

7. Mitochondrial remodeling is impaired in aging

Mitochondria are highly mobile and remarkably plastic
organelles that continuously undergo organellar turnover and
morphology remodeling. Evidence is accumulating that
mitochondrial dynamic remodeling are sensitive to various
physiological and pathological stimuli, and mitochondrial
remodeling are no epiphenomena, but central to cell function
and survival.
Mitochondria are able to change their architecture from
individual structures to complex tubular networks through
fusion and fission. It has been established that this dynamic
structure is regulated by proteins controlling fission, such as
mitochondrial fission 1 protein (Fis1) and dynamin-related
protein 1 (Drp1), and fusion, such as mitofusin homologs
Mfn1/2 and Dynamin-like 120 kDa protein OPA1. Genetic
defects in the proteins involved in mitochondrial fusion and
fission lead to severely altered mitochondrial shape, loss of
mtDNA integrity, increased oxidative stress and apoptotic cell
death.33 In healthy cells, mitochondrial fusion provides
a synchronized internal cable for translocating metabolites and
intramitochondrial mixing during biogenesis, whereas

mitochondrial division facilitates the equal distribution of

mitochondria into daughter cells during mitosis and allows the
selective degradation of damaged mitochondria through
autophagy. However, it has become increasingly clear that
these protective mechanisms are markedly impaired in aging
and that faulty mitochondrial dynamics might be involved in
the aging process. Crane et al.34 reported that the expression of
Mfn2 and Drp1 genes is reduced in the skeletal muscle of
aging individuals. Furthermore, Using RNA interference to
reduce Fis1 in mammalian cells, it was found that mitochondria become enlarged and flattened and these morphological
changes are correlated with increased a-galactosidase activity,
a marker of cell senescence, and decreased mitochondrial
membrane potential, increased ROS generation, and DNA
damage.35
The constant renewal of mitochondria is crucial for maintaining healthy mitochondria with age. Accurate organellar
turnover requires the coordination of two key cellular
processes: mitochondrial biogenesis and selective degradation
(autophagy). The capacity for mitochondrial biogenesis
diminishes with age and this is an important parameter in the
mitochondrial dysfunction associated with aging.36 If
biogenesis is affected, it is reasonable to expect that mitochondrial turnover must be slower and the accumulation of
modified lipids, proteins and DNA must also increase, further
aggravating the situation resulting from the deficient activity
of aged mitochondria. The precise reason for the decrease in
the rate of mitochondrial biogenesis during aging is currently
unknown. However, it seems that both extra- and intracellular
regulatory factors of mitochondrial biogenesis are implicated.
In fact, the pleiotropic signaling of mitochondria, H2O2 and
NO diffusion to the cytosol is modified in aged animals and
contributes to the decreased mitochondrial biogenesis in old
animals.37
Autophagy appears to decline with age, and several key
players in the autophagic pathway (e.g., ATG5 and ATG7)
show decreased expression in the brains of aging individuals.
Stimulation of autophagy can increase the healthy lifespan in
multiple model organisms, including mice and primates.38
Autophagy can mitigate inflammatory reactions through
several mechanisms. Recently, it was found that autophagy
can inhibit NLRP3 activation by removing permeabilized or
ROS producing mitochondria.39 Because pathological aging is
accompanied by chronic inflammation, these anti-inflammatory effects of autophagy may mediate additional health
benefits. Mitochondrial dynamics are also important for proper
organellar turnover in that they affect mitochondrial degradation pathways. Fis1 expression was found to be reduced in
abnormal mitochondria, whereas overexpression of this
protein blocked the senescence-related phenotype and maintained cells in a proliferating state. The link between fission
proteins and mitochondrial turnover has also been illustrated
in neuronal cells, in which Fis1 was shown to activate autophagy and the selective degradation of depolarized
mitochondria.40
In summary, aging compromises the remodeling of mitochondria by disrupting the homeostatic regulation of

Mitochondrial redox metabolism in aging and exercise

71

mitochondrial fusion and fission pathways. The presence of

fragmented mitochondria owing to a decline in fusion and/or
an increase in fission events can compromise mtDNA integrity,
mitochondrial structural and functional complementation, and
mitochondrial biogenesis, all of which can lead to mitochondrial dysfunction. Conversely, the formation of enlarged
mitochondria as a result of decreased fission and/or increased
fusion events can diminish mitochondrial turnover by
impairing autophagy and biogenesis, leading to the accumulation of damaged mitochondria in aged cells. In both cases,
the abnormal mitochondria are unable to fulfill their lifesustaining roles. Therefore, age associated alterations in
mitochondrial fusion and fission dynamics might play a causative role in mitochondrial remodeling impairment and
increase susceptibility to cell death in response to various
types of stress during progressive aging.

old rats. It was shown that an age-associated lack of expression of PGC-1a in response to not only training but also to
cold exposure or triiodothyronine. Recently, Farhoud et al.48
reported that chronic oxidative stress suppresses the gene
expression of PGC-1a through activation of the ubiquitinproteasome system, thereby inhibiting its downstream mitochondrial biogenesis and metabolic gene expression programs.
In addition, chronic inhibition of NF-kB via the pharmacological agent pyrolidine dithiocarbamate (PDTC) was found to
increase PGC-1a level suggesting inactivity might downregulate PGC-1a due to elevated NF-kB activity.49 It could be
presumed that aging related inflammation and subsequent
activation of NF-kB could negatively influence the expression
of PGC-1a and its effect on mitochondrial biogenesis. These
results showed that loss of mitochondrial biogenesis during
aging may be due to a lack of induction of PGC-1a.

8. PGC-1a, mitochondrial remodeling, and aging

9. Mitochondrial remodeling, ROS, and exercise

Despite the complexity of the various signaling pathways

that regulate mitochondrial remodeling, they all seem to share
proliferator-activated receptor-a coactivator 1a (PGC-1a)
family of transcription factors. PGC-1a appears to act as
a master regulator of energy metabolism and mitochondrial
remodeling by coordinating the activity of multiple transcription factors. PGC-1a strongly co-activates NRF-1, which
is an intermediate transcription factor to stimulate the
synthesis of mitochondrial transcription factor A (Tfam).
Tfam can be considered the most important mammalian
transcription factor for mtDNA because it stimulates mitochondrial DNA transcription and replication. Recently, Aquilano et al.41 showed that PGC-1a and Sirt1 not only colocalize
with the nuclear, but also interact with Tfam in the mitochondria to form multiprotein complexes. In addition, PGC-1a
has been shown to be involved in mitochondrial network
remodeling through controlled fusion and fission. H2O2-stimulated Mfn1/2 expression was dramatically attenuated in
PGC-1a knockout C2C12 muscle cells,42 whereas PGC-1a
over-expression markedly enhances Mfn2 mRNA and protein
levels in cultured muscle cells.43 PGC-1a also stimulates the
transcriptional activity of the human Mitofusin 2 (Mfn2) gene
promoter, which requires the integrity of the nuclear hormone
estrogen-related receptor ERRa-binding element.44 Finally,
PGC-1a controls most of the enzymes and proteins affecting
oxidanteantioxidant balance such as UCP2/UCP3, MnSOD,
and GPx expression.45
Kang et al.46 reported that aged (24 months) rats had
significantly lower gene expression in the PGC-1a signaling
pathways, shown by decreased mRNA and protein contents for
PGC-1a, Tfam and cytochrome c compared to young
(4 months) rats. Furthermore, phosphorylation of the upstream
enzymes AMP kinase and p38 MAPK, as well as cAMP
response element nuclear binding, was down-regulated. Derbre et al.47 studied mitochondrial biogenesis and PGC-1a
expression in old rats and compared them with PGC-1a
knockout mice. They found a striking similarity between the
response to exercise training in PGC-1a knockout mice and in

The term Hormesis has been adopted to explain how a mild

oxidative stress associated with exercise can result in favorable
adaptations that protect the body against more severe stresses
and disorders derived from physical stress or other etiological
origin. The typical reaction to ROS can be described by a bellshaped curve: low concentrations have a stimulating effect
(signaling, receptor stimulation, and enzymatic stimulation),
while a massive level of ROS inhibits enzyme activity and
causes apoptosis or necrosis. Recent evidence suggests that
suppression of ROS production fails to extend lifespan in
worms and may even decrease lifespan in humans, presumably
due to the reduction of the ROS signaling, which seems to be
important for different cellular processes.50,51 It was suggested
that exercise could increase the formation of ROS to a level
that may induce significant, but tolerable, damage that can in
turn induce beneficial adaptations. Short-term ROS production
is apparently important in prevention of aging by induction of
a process named mitohormesis and redox signaling.52 This
process seems to be particularly important for the health effect
of exercise.
Recently, we studied the effect of aging and exercise on
mitochondrial remodeling in skeletal muscle because it contains
mainly post-mitotic cells and because of the importance of this
organ in aging (Fig. 1). Mfn1 and Drp1 protein were unchanged
at 15 months. Mfn1 was decreased 39% at 20 months vs.
5 months ( p < 0.05), whereas Drp1 showed w70% ( p < 0.01)
increase at 20 months vs. 5 months. Compared with control
group, 12-week treadmill training resulted in an increase in
Mfn1 in 15 months ( p < 0.01), whereas in a small decrease in
20 months ( p < 0.05) (Fig. 1A). In contrast to Mfn1, compared
with control group, training resulted significantly increased in
Drp1 protein both at 15 and 20 months vs. 5 months ( p < 0.01
and p < 0.05, respectively) (Fig. 1B). Furthermore, cytochrome
c oxidase subunit IV (COXIV), the marker of mitochondrial
biogenesis, showed a 30% increase at 20 months vs. 5 months
( p < 0.05). Compared with control group, training increased
COXIV content by w30% at 5 and 15 months old rats
( p < 0.05), and by w20% at 20 months old rats ( p < 0.05)

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H. Bo et al.

Fig. 1. Western blot analysis of Mfn1 (A), Drp1 (B), COXIV (C), and mitochondrial ATP synthase activity (D) in rat skeletal muscle of 12-week treadmill trained
5-, 15-, and 20-month-old rats and controls. yp < 0.05, yyp < 0.01: trained group compared with control group at various ages; *p < 0.05, **p < 0.01: in control
group, compared with 5 months; #p < 0.05, ##p < 0.01: in trained group, compared with 5 months.

(Fig. 1C). We also measured ATP synthase activity as indication

of mitochondrial energy production. ATP synthase activity was
unchanged at 15 and 20 months vs. 5 months, whereas training
increased its activity at various ages, 5-, 15-, and 20-month-old,
compared with control group ( p < 0.01, p < 0.05, and p < 0.05,
respectively) (Fig. 1D). Taken together, our data suggest that
regular exercise training stimulates mitochondrial biogenesis,
a rejuvenation of the mitochondrial network via fission and
fusion, and an improved efficient of mitochondrial energy
transfer. All of these properties, working in conjunction with one
another, would improve the overall functionality of mitochondria in aged cells (Zhang et al., unpublished data).
Gomez-Cabrera et al.53 demonstrated a reduction in mitochondrial biogenesis and protein quality control factors
compared to placebo control, following supplementation of
vitamin C during a 6-week endurance protocol in rats. It is
clearly shown that exercise induced mitochondrial mass
remodeling is a redox-sensitive process. Recently, we found
that exercise induced mitochondrial ROS may contribute to
the rapidly alteration in mitochondrial fusion/fission protein
expression.54 Koopman et al.55 reported that vitamin E
supplementation reduced ROS production and restored aberrant mitochondrial morphology in fibroblasts with mitochondrial complex I deficiency, suggesting that ROS is involved in
controlling mitochondrial shape in these cells. Using a nonapoptotic concentration of H2O2 in long-term treatment,

originally long and interconnected mitochondrial tubules were

transiently shortened and weakly fragmented.56 In addition,
low doses H2O2 in a transient exposure induce mitochondria
hyperfuse and form a highly interconnected network in cells.57
Based on the above evidence it can be concluded that ROS
induce remodeling of the mitochondrial network depending on
the concentration of ROS and duration of the treatment. ROS
is also an important signal for induction of autophagy. Starvation-induced autophagy can be suppressed by antioxidants
suppressing the well-known prosurvival function of starvationinduced autophagy.58 Recently, it is demonstrated that exercise
is a potent inducer of autophagy, and that acute and chronic
exercise enhances glucose metabolism in mice capable of
inducing autophagy but not in autophagy-deficient mice.59
Several studies have shown that an acute bout of endurance
exercise and stimulated muscle contraction can upregulate PGC1a and activate mitochondrial protein synthesis and proliferation.60 Furthermore, repeated exercise bouts (exercise training)
could result in accumulation of PGC-1a, NRF-1, and Tfam
protein levels.61 These observations were thought that PGC-1a
plays an important role in mediating mitochondrial adaptation to
exercise, such as elevated respiratory activity, increased
expression of Krebs cycle enzymes, promoted endogenous
antioxidant defense capacity, enhanced fatty acid oxidation, and
mitochondrial morphological changes. Garnier et al.62 demonstrated in healthy humans that VO2peak was dependent on

Mitochondrial redox metabolism in aging and exercise

coordinated expression of PGC-1a, which was lineally correlated to Mfn2 and Drp1 levels in human leg muscle. In C2C12
skeletal muscle cells, increasing the production of ROS by
incubation with 300 mM H2O2, results in increased PGC-1a
promoter activity and PGC-1a mRNA expression requiring
the activation of AMPK.63 Ristow et al.64 demonstrated
exercise-induced oxidative stress elevated PGC-1a and
PGC-1b expression and ameliorated insulin resistance, which
were precluded by antioxidants supplementation.
10. Conclusion
A vast body of data has accumulated linking mitochondrial
redox metabolism to the aging process. Therefore, the accumulation of oxidative damage can be decreased either by
lowering the generation of ROS (caloric restriction) or by
regular exposure to a small amount of ROS (such as mild
exercise) that could result in slight oxidative damage, which
then leads to up-regulation of antioxidant systems and
amelioration of mitochondrial remodeling. Signal transduction
by coordinated action of PGC-1, NF-kB, and MAPK (p38)
may be potential regulators in these events. These products
include (1) antioxidant enzymes (e.g., MnSOD, GPx, GCS);
(2) molecules controlling metabolic status and thus ROS
production, (e.g., UCPs, enzymes in fatty acid and glucose
metabolism); (3) transcription factors for mitochondrial
biogenesis (e.g., PGC-1a, NRF-1, Tfam), and (4) a wide range
of proteins that could influence mitochondrial dynamic
remodeling, such as mitochondrial fusion/fission and autophagy proteins. Exercise training is a useful and inexpensive
intervention for preventing aging and degenerative disease, in
which mitochondrial redox metabolism plays a key role.
Acknowledgments
This work was supported by research grants from the National
Natural Science Foundation of China (No. 31110103919,
31200894, 31000523, 30771048, 30470837, 31071040, and
30270638), Tianjin Municipal Sci-tech-innovation Base Project
(No. 10SYSYJC28400), Tianjin Science and Technology
Planning Project (No. 12JCQNJC07900), and General Administration of Sport of China Basic Project (No. 10B058).
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