Sports Medicine and Health Science 1 (2019) 24–32

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Sports Medicine and Health Science

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High intensity interval training and molecular adaptive response of

skeletal muscle
Ferenc Torma a, Zoltan Gombos a, Matyas Jokai a, Masaki Takeda b, Tatsuya Mimura c,
Zsolt Radak a, *
a
Research Center of Molecular Exercise Science, University of Physical Education, Budapest, Hungary
b
Faculty of Health and Sports Science, Doshisha University, Kyotanabe, Japan
c
Faculty of Sport and Health Sciences, Osaka Sangyo University, Osaka, Japan

A R T I C L E I N F O A B S T R A C T

Keywords: Increased cardiovascular fitness, VO_ 2max, is associated with enhanced endurance capacity and a decreased rate of
High intensity interval training _ 2max and endurance
mortality. High intensity interval training (HIIT) is one of the best methods to increase VO
Cellular adaptation
Molecular pathways
capacity for top athletes and for the general public as well. Because of the high intensity of this type of training,
Redox signaling the adaptive response is not restricted to Type I fibers, as found for moderate intensity exercise of long duration.
Maximal oxygen uptake Even with a short exercise duration, HIIT can induce activation of AMPK, PGC-1α, SIRT1 and ROS pathway as well
Mitochondrial biogenesis as by the modulation of Ca2þ homeostasis, leading to enhanced mitochondrial biogenesis, and angiogenesis. The
present review summarizes the current knowledge of the adaptive response of HIIT.

Introduction oxygen uptake (VO _ 2max), introduced by the Nobel laureate Archibald
Vivian Hill, become widespread only after the Second World War.
High intensity interval training (HIIT) emerged several decades ago in Depending on the applied protocol, the exercise resembled endurance
response to the need for new training techniques for athletic events of moderate intensity continuous training (MICT) or a strength-speed
high intensity and often also of long duration. In the 1920s the legendary training adaptation. For an in depth historical review the reader is
runner “Flying Finn” Paavo Nurmi started to introduce “interval training” directed to the review of Veronique Billat.2
sessions to his annual training cycle.1 In the Nordic athletic community It became clear that high intensity exercise is a fundamental tool for
“natural” interval training gained popularity, was named Fartlek and was improving peak athletic performance. It is impossible to reach the
formalized by G€ osta Holmer in 1937. The Fartlek method originally was required physiological limit of athletic performance during competitions
developed for cross country, multi-terrain runners and consisted of a without challenging the musculoskeletal and cardiovascular systems by
continuous distance run in which very high (sprints, or uphill runs) and vigorous exercise sessions. As early as 1986 Cox and colleagues showed
low velocity running periods were integrated. In the 1950s interval that after seven weeks working at 90% or higher intensity exercise
training methods started to infiltrate several European athletic training induced left ventricular morphology changes in previously sedentary
programs. It became prominent probably because the four-time Olympic individuals.6 HIIT is shown to increase aerobic capacity even more effi-
gold medalist Emil Zatopek (Czeckoslovakian) and other eminent run- ciently than volume-based endurance training protocols.7 The time spent
ners like Vladimir Kuts (Russian), Gordon Pirie (British) or Sigfried with doing physical exercise is also an important factor, not only for
Hermann (German) used this training method effectively during their nonprofessional athletes but for professionals, as well. Recently
preparation.2 Some of the first scientific papers which described HIIT in time-efficiency became a pinnacle aspect for athletes to improve diverse
detail were published in the late 50s-early 60s by Roskamm and Reindell3 skill sets. HIIT induces comparable chronic physiological adaptation with
as well as Per Oløf Astrand.4 However, HIIT research gained full attention less expenditure of time than classical endurance programs,8 and the
in the 1970s.5 Early protocols used the average velocity or the velocity recovery time after HIIT appears to be shorter than after moderate in-
corresponding to the personal best time of the distance of interest as tensity exercise of long duration.9 Moreover, some authors claim that
reference instead of the maximal aerobic capacity. The use of maximal HIIT is a superior method compared to aerobic training, in order to

* Corresponding author. Alkotas u. 44, Budapest, H-1123, Hungary.

E-mail address: [email protected] (Z. Radak).

https://doi.org/10.1016/j.smhs.2019.08.003

Available online 11 September 2019

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F. Torma et al. Sports Medicine and Health Science 1 (2019) 24–32

improve diverse physiological functions.10–12 The question that needs to same time creatine phosphate, glucose 6-phosphate and ATP gradually
be answered is “what are the differences in the adaptive response to HIIT decreased in the quadriceps muscle.
and MICT”. When one attempts to evaluate the actual metabolic features of SIT or
In this review we focus on investigations that applied 85–90% [taking HIIT, in respect to MICT, a major problem emerges, because the volume
peak power output (PPO) or PVO _ 2max vVO_ 2max] or higher loads during and the energy usage are not always matched. It is obvious that higher
HIIT. This range of intensity is suggested by Wisløff et al.13 who reviewed power output transiently increases the energy demand but low volume or
the literature on interval training and its cardiac benefits. The duration of the total work performed during HIIT/SIT exercise sessions makes it
the high intensity load is also a matter of debate. The protocols in the difficult to access the effects of prior exercise intensity and modality on
following sections will be referred to as HIIT if one session does not metabolic substrate turnover immediately after exercise or with recov-
exceed 4–5 min during the total exercise time. Sprint interval training ery. Lactic acid production and post exercise levels after exercise are
(SIT) will be referred to as “all out” maximal effort (greater than generally higher after HIIT20 compared to MICT. In a comparative study,
_ 2max) if the duration is less than 1 min per each sprint. Our nomen-
PVO where long and short interval HIIT-s and MICT were matched for the total
clature will follow the terminology used by MacInnis and Gibala14 in duration, mean power showed significantly higher levels of lactic acid in
their review. The present paper aims to discuss the molecular adaptive peripheral blood after long duration, but were less pronounced after a
response of HIIT and emphasizes the differences from moderate intensity short interval HIIT program.21 It is not easy to estimate how metaboli-
exercise with long duration. cally challenging a given HIT protocol is, since, as mentioned above,
What is high intensity interval training? besides the intensity level, the work to release ratio and the release in-
Interval training can be defined simply as discontinuous, periodic tensity (is it a passive or an active rest period?) can affect the metabolic
(demanding) exercise loads separated by periods of recovery. However, burden. A suggested method to estimate the SIT or HIT training meta-
the intensity level which defines an exercise as “high intensity” varies in bolic demand is to consider the five-minute lactic acid increment from
different research publications. There is no accurate consensus threshold the start of the exercise. Peripheral lactic acid may not be the perfect
from which point the training load is categorized as ‘close to the maximal indicator of the exercise intensity since it can be affected by dietary
effort’. In the literature the terms “high intensity” or “vigorous” are used status22 and it is not perfectly, but reflects the overall anaerobic envi-
in a fairly diverse manner ranging from an effort as low as 65% of the ronment in the exercising skeletal muscle23 and it provides sufficient
peak power output15 up to 170% of the measured power at VO _ 2max.16 information about the peripheral metabolic environment. This finding
Even the dimension of reference intensity can be different in publications highlights the main physiological divergence between HIIT and MICT.
_ 2max%) or ve- During MICT the exercise is conducted in a physiological steady state, i.e.
since peak power output percentage (PPO%), power (PVO
metabolic processes can fulfill the energy demand of skeletal muscle with
_ _
locity (vVO2max%) percentage of the VO2max, maximal heart rate per-
no significant temporal metabolite accumulation. However, as intensity
centage (HRmax%) and critical/maximal velocity percentage (vVmax%)
increases and the maximal lactate steady state is approached, the
are all commonly used, but, vary greatly. Using different reference di-
dominance of oxidative processes decreases, lactate accumulation takes
mensions can be problematic because the same numerical values may
place and, if it is too rapid, it results in quick exhaustion. In HIIT, energy
indicate a different metabolic demand with a different dimension.15 Most
need is only transiently increased in the tissue and the oxygen deficit can
frequently, intensity levels are referred to the work load corresponding to
be countered during the resting phase, but the “metabolic perturbation”
the 90% of maximal aerobic capacity (or 90–95% of the peak HR) or
is a function of the intensity level and the recovery time. Because of the
maximal effort. Buchheit and Laursen noted nine characteristics which
relatively short, high intensity bursts, the local metabolic capacities (CP,
can precisely describe HIT exercise17 including interval intensity and
ATP, and mitochondrial enzymes) are able to cope with the increased
duration, relief interval intensity and duration, exercise modality, num-
demand, and the loss can be countered during recovery. Therefore, the
ber of reps and series, and finally between series recovery duration and
accumulation of lactic acid is slower and the onset of fatigue can be
intensity.
prolonged, even at high levels of intensity.
According to their publications on HIIT, protocols can be divided into
In general, HIIT/SIT changes in substrate utilization or mobilization
two basic groups:
are believed to be steeper than during MICT. Lactate24 and plasma
glucose levels25,26 increased, PCr/(Cr þ PCr) ratio25 decreased during
(i) High intensity with relatively higher volume, 3–5 min with “near
high intensity exercise, but post exercise glycogen content can be
maximal” effort (~80–90% intensity), separated by resting pe-
similar25,27 when compared to MICT. However, the chronic HIIT/SIT
riods (but one can find protocols using more than five-minute
interventions seem to increase the resting intramuscular glycogen pool.28
sessions).
The discrepancies, in the case of acute exercise, probably can be
(ii) The other form is “the all-out” or “until exhaustion” type of ex-
explained by the different energy expenditures between HIIT and MICT.
ercises often referred to as SIT. In the case of SIT, one sprint rarely
Intensive exercise has a major impact on the systemic sugar homeostasis
exceeds a 30 s time interval and training intensity is termed to be
since as high as 80% of the glucose can be transposed into the tissue when
“supramaximal” while exercise peak power output (PPO) levels
insulin is present.29 The ATP turnover rate can be 100- fold higher during
exceed the power value recorded during VO _ 2max measurements.
exercise than at rest.30 After a training period of six weeks at 90% VO _ 2
peak, 3 d/wk, glycogenolysis and lactate accumulation were decreased31
HIIT and SIT are usually associated with continuous cyclic move-
during the test phase and the protein content of MCT (monocarboxylate
ments like running, cycling, and rowing, which can induce morpholog-
transporter) 1 and MCT 4 lactate transporters, fatty acid transporters
ical changes such as a conversion of fiber type morphology, increased
FAT/CD36, FABPpm, GLUT4 glucose transporter, maximal activities of
fiber type area in fast twitch fibers etc.18
mitochondrial aspartate aminotransferase, β-hydroxyacyl CoA dehydro-
genase (β-HAD) and total pyruvate dehydrogenase increased signifi-
Molecular effects of high intensity interval training cantly, as a result of training.
Due to the different metabolic rates of HIIT and MICT there is also a
The pioneering work of Karlsson and Saltin19 examined the molecular difference in metabolite production. The lactate levels in blood increase
effects of HIIT in skeletal muscle. Subjects were exposed to five cycling during the 5 min during exhaustive exercise32 but when longer duration
sessions with 1 min supramaximal load ~120% of VO _ 2max followed by applied, lactate accumulation plateaued.33 Needless to say, the accu-
5 min of passive rest. Oxygen uptake and lactate levels increased signif- mulation of lactate is dependent on the exercise intensity.34
icantly during the exercise, but, interestingly enough, after the first burst In one elegant study, ten well-trained males completed a single bout
of exercise, lactate values did not increase as the load progressed. At the

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F. Torma et al. Sports Medicine and Health Science 1 (2019) 24–32

of isoenergetic HIIT or MICT in a randomized and counterbalanced arteriovenous difference, HIIT positively affects all the major cardiac
order.24 The carbohydrate oxidation was higher, whereas fat oxidation markers (cardiac output, maximal HR, stroke volume).45 It is important
was lower during HIIT vs. MICT. From the 49 compounds measured by to establish some common reference point where the differential adap-
targeted metabolic analysis after exercise from plasma samples, 13 tation pattern of HIIT/SIT and MICT can be studied, because the energy
changes were characteristic of HIIT and only five of MCIT. Tricarboxylic demand, the distance covered, or the work being done may be incom-
acid intermediates (citric, aconitic and succinic acid) increased to a parable. The most widely used methods are the isocaloric/isoenergetic
greater extent after HIIT compared to MICT. Amino acid alanine levels (work-matched) or volume/duration matched study designs. When these
elevated only during HIIT, but the two BCAA compounds, valine and corrections are applied, the universal superiority of HIIT becomes less
leucine, were significantly lower after MICT in plasma samples compared unequivocal46,47 (Fig. 1.). But generally, it seems that higher intensity
to the pre-exercise values. Despite the valuable information about exercise has the capacity to induce more pronounced beneficial changes
HIIT-induced metabolic perturbation, the biological role of plasma TCA in physiological and molecular markers.48
and other intermediaries is rather confusing since these metabolites play The large energy demand of HIIT results in significant increases in
primal role in the working skeletal muscle. cellular ADP/ATP and AMP/ATP ratios which can activate AMPK, one of
the most important energy sensors of the cell.49 AMPK is made up from α,
β, and γ subunits. If ATP concentration decreases and AMP levels increase
HIIT and MICT training impact on skeletal muscle oxidative capacity as a result of muscle contraction or the lack of energy influx, then AMP
bounds to Bateman domain (CBS domain) of γ-subunits as the first event
The different adaptations to exercise stimuli of skeletal muscle fiber in the activation of mammalian AMPK.50 Several studies showed that
types may help to answer the question: why is HIIT exercise capable of AMPK phosphorylation in skeletal muscle positively correlates with the
increasing aerobic capacity in a relative short time? Performing physical exercise intensity25,41 and duration.51 AMPK phosphorylates several
exercise at high intensity activates type II fiber contraction, obeying the crucial enzymes involved in regulation of lipid and protein metabolism
size principle.35 Data suggest that mitochondrial content of Type I fibers and glucose transport.52 The heterotrimer combination may provide an
can be increased by low intensity exercise of long duration, while in case important way of adaptation to different exercise stimuli. Hence the
of Type II fibers, mitochondrial biogenesis is induced by high intensity different association of α, β and γ isoforms (α1, α2, β1, β2, γ1, γ2, γ3) can
_ 2max is dependent on arterio-venous oxygen differ-
exercise.36 Since VO differ in fiber types.53 The activity of α subunits and increased α2 acti-
ences, which is influenced by mitochondrial content, it is easy to un- vation (as assessed by isoform specific immunoprecipitation) levels were
derstand why HIIT-induced mitochondrial biogenesis is associated with found after 20 min of exercise only if the intensity was higher (i.e. 20
enhanced VO _ 2max. In addition, the induction of HIF137 and VEGF37,38 min. cycling at 50% or 70% PVO _ 2max),54 nevertheless the α1 activity
have been reported, and this adaptive response may be also important to levels were unaffected (the glycogen and PCr level dropped after the 70%
create greater arterio-venous oxygen differences and VO _ 2max. In contrast, bout). In a similar study isoform-specific AMPK activity measured by
after eight weeks of MICT39 a superior increase in capillary density was sequential immunoprecipitation after a single but of HIIT or MICT. The
detected when compared to HIIT with similar total energy expenditure. increase in activity of the α2β2γ3 structure/construction was 27- fold in
Data with similar vascular adaptations27 is also known. the HIIT vastus lateralis samples compared to the 12- fold in MICT.25 On
Tan and his colleagues showed that when sedentary young woman the other hand, independently of exercise trial, α1β2γ1 assembly activity
conducted a six-week low volume HIIT training (~90% HRmax) the decreased with exercise and α2β2γ1 AMPK activity remained unchanged.
intervention elevated the mitochondrial COX IV in both type I and type II Both exercise interventions elevated AMPK (Thr172), ACC (Ser221),
fibers.40 Indeed, there are numerous studies about HIIT or SIT that have TBC1D1 (Ser231) and TBC1D4 (Ser704) phosphorylation similarly in
demonstrated increased mitochondrial biogenesis41,42 or elevated total muscle lysates. However, phosphorylation of AMPK (Thr172)
oxidative capacity.43,44 It is also known that if HIIT and MICT are applied increased by 184% in type II fiber and only in the HIIT group. Similar
in an isocaloric manner, besides the improving effect on the observations have been made by others,55,56 underlining the role of the

Fig. 1. The energy demand of exercise with different intensities. Thesuggested energy cost of MICT, HIIT and SIT. The discrepancies of the results in various
studies are due to the different energy costs of exercise training. Matched energy or power cost are required to valid evaluation.

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F. Torma et al. Sports Medicine and Health Science 1 (2019) 24–32

α2 isoform in exercise dependent AMPK regulation. The available in- time) with 4 min rest between bouts in recreationally active subjects
formation indicates that AMPK isoform-specific activity may play an caused the fragmentation of ryanodine receptor type 1 (RyR1) 24 h after
important role in HIIT-induced metabolic adaptation, however the ada- the exercise stimuli.58 After SIT, only 15% of the RyR1 remained intact
tive response to HIIT is very complex (Table 1). and ~375, 80, and 60 kDa fragments were detected. However, this
response was not present in elite well trained athletes nor in athletes
Calcium mediated regulation preformed marathon race. The authors suggest the involvement of
reactive oxygen species (ROS), since superoxide dismutase 2 (SOD2) and
Another important event in skeletal muscle functional activity is the catalase (CAT) expression are at least two-fold higher in vastus lateralis
muscle contraction-initiated calcium release from the sarcoplasmic re- muscle of elite athletes at baseline, indicating better antioxidant capac-
ticulum by a ryanodine receptor-mediated mechanism.57 Inside skeletal ity. Endogenous antioxidants59,60 and Ca2þ handling61 are associated
muscle cells, calcium sensitive proteins evolve to mediate metabolic and with enhanced mitochondrial function and recurrent high Ca2þ, AMP
electrophysiological pathways, during physical exercise. Place and col- and ROS levels during high intensity exercise may contribute to mito-
leagues showed that a 30s all-out cycling exercise (3 min total exercise chondrial biogenesis by activating peroxisome proliferator-activated

Table 1
HIIT induced adaptive response on humans.
Reference Population characteristics (n, M/F, Training period Exercise details Main physiological and molecular changes
_ 2max)
age, VO
47
6, 6/0, 25  2.9 yr, 55.5  1.3 mL/ Single bout, HIIT: ~40–45 min 2 min at 90% and 2 min at Postprandial fat oxidation was increased similarly in
kg/min cycling 25% VO_ 2peak.MICT: ~60 min of at 50% both exercise groups with a higher increase in HIIT.
_ 2peak.
VO
116
10, 10/0, 20 1yr., 52  7 mL/kg/ Single bout, _ 2maxand 3-min at
HIIT: 3-min bouts at 90% VO Heart rate, RPE, and blood lactate was significantly
min running 50% VO _ 2max.
_ 2max. MICT: 50 min at 70% VO higher in HIIT compared with MICT. Phosphorylation
of AMPK Thr172 and p38MAPK Thr180/Tyr182
increased after exercise with no difference between
exercise protocols. Muscle (vastus lateralis) PGC-1α
mRNA content increased after hrs. of exercise with no
difference between protocols. PGC-1α protein content
was not changed at any time during the HIIT or MICT
trials.
24
10, 10/0, 33.2  6.7yr., 4.8  0.3 L/ Single bout, _ 2max
HIIT: 10  4 min cycling at 81.6  3.7% VO Plasma lactate, adrenocorticotrophic hormone,
min (~61 mL/kg/min) cycling and 2 min with at 50 W (11.4 0.9% peak power cortisol, and growth hormone were all higher
_ 2max for a time
output). MICT: cycling at 65% VO immediately after HIIE vs. MICT. Plasma
corresponds to total HIIT work. norepinephrine and interleukin-6 increased similarly.
Plasma insulin decreased during recovery in both
HIIE and MICE.
117
44,0/44, ~25yr.,~29 mL/kg/min 3 times a week for _ 2max and
HIIT: 40 min with 1 min at 80–90% VO HIIT was superior in improving norepinephrine,
16 weeks, 2 min at 50–60% VO_ 2max. MICT: 40 min at endothelin-1 (ET-1) and (nitrite/nitrate) NOx
running/walking 60–70% VO _ 2max. response to exercise than MICT. HIIT and MICT were
similarly effective in improving ABP and insulin
sensitivity. HIIT was superior in improving
cardiorespiratory fitness.
118
16, 0/16, HIIT: 20  1 yr. MICT: 5 weeks 3 days HIIT: 120% (week 1), 130% (weeks 2 and 3) and In HIIT group there was a significantly greater
19  1 yr, HIT: 43.7  6.8 mL/min/ per week, cycling 140% (weeks 4 and 5) of the lactate threshold, improvement in vastus lateralis muscle buffer
kg, MICT: 42.1  7.2 mL/min/kg 2 min duration, with 1 min recovery. MICT: 80% capacity (βm in vitro) than the MICT group. VO_ 2peak
(week 1), 90% (weeks 2 and 3) and 95% (weeks 4 increased in both group similarly.
and 5) of the lactate threshold. Work matched to
HIIT.
119
40, 40/0, 24.6  3.8 yr, 8 weeks 3 days HIIT: 4  4 min at 90–95% HRmax with 3min _ 2max and stroke volume
After HIIT and SIT VO
~55–60 mL/min/kg per week, active recovery at 70% HRmax. SIT: 47  15-s at increased significantly
running 90–95% HRmax with 15 s active recovery at 70%
HRmax. MICT: at 70% HRmax for 45 min.
120
43, 17/26, 55–79 yr, 8 weeks 4 days HIIT: 4  4 min at 85–95% HRpeak with 3 min _ 2peak, ejection fraction and insulin resistance
VO
~23,1–25,9 mL/min/kg per week, all- active recovery at 65–75% HRpeak. MICT: (HOMA-IR) improved in HIIT.
extremity 32 min at 65–75% HRpeak.
ergometer
46
10, 10/0, 23  1 yr, 46  2 mL/kg/ 2 week 6 session, HIIT: 4  5 min at 65% Wmax and 2.5 min CS maximal activity and mass specific O2 flux
min One-leg cycling recovery with 20% Wmax. MICT: 30 min with oxidative phosphorilation capacities in HIIT vs. MICT.
50% Wmax In whole muscle, the COXIV, NDUFA9 and mitofusin
2 (MFN2) increased similarly in both groups.
121
9,n.d.,20–28 yr, ~25.7–61.3 mL/ 7–8 week 3 days/ _ 2max
HIIT: 5  4 min 2 min recovery, 101% of VO _ 2max elevated only in the MICT group. SDH activity
VO
kg/min week, cycling matched by W. MICT: average 27 min, 79% of increased in both group, but no difference found in
_ 2max.
VO PFK levels.
122
26, 10/16, obese,41  9 yr, 18 session 3/ 4HIIT: 4  4 min at 85%–95%HRmax with 3 min _ 2max increased in the 4HIIT group compared to the
VO
~31–36.2 mL/kg/min week, running recovery at 70% HRmax. 1HIIT: 10  1 min 90% other two groups. Calculated stroke volume increased
HRmax with active recovery time n.d. MICT: only in the 4HIIT group. Muscle CS activity and TTE
45 min at 70% HRmax. (time to exhaustion) improved in all exercise group
with a difference between 4HIIT and MICT in TTE.
123
17, 17/0, ~24.6  3 yr, 8 week 3 day/ HIIT: 10  2 min at 105% with 2min recovery. _ 2max and maximal exercise ventilation (VEmax)
VO
3046–3757 mL/min week, cycling 1MICT: 55 min of continuous exercise at 50%. increased in all protocol with a higher increase of
_ 2max; 2MICT: 35 min at 70% VO
VO _ 2max; VEmax in HIIT to other MICT groups.
124
13, 13/0, 19–25yr.,37–54 mL/kg/ 4 week 5 One leg SIT: 20–30 bouts of 40–50 s all out efforts Muscle SDH activity increased in both exercise
min workout/leg/ with 60–90 s recovery. One leg MICT: groups.
week, cycling 30–50 min at 75% of VO_ 2max

27
F. Torma et al. Sports Medicine and Health Science 1 (2019) 24–32

receptor-gamma coactivator (PGC)-α and the downstream nuclear res- COXIV protein levels more in rat soleus and rectus femoris muscle than
piratory factor 1–2 (NRF1- NRF2) systems.62 However, it is not clear why MICT.78 It is interesting, that PGC-1a can directly be involved in the
individuals with various levels of fitness respond differently to HIIT-like transcriptional control of lactic acid production79 by reducing the
exercise stimuli. One possible explanation for this phenomenon is that expression of LDH A and one of its regulators, the transcription factor
training causes optimal biological responses at a distinct range of in- myelocytomatosis oncogene. Fiorenza et al., in order to “induce distinct
tensity for each person and, presumably, it follows a hormetic trajectory metabolic perturbation” in the myocellular environment, conducted
in groups with different training backgrounds.9,63 repeated sprints (18  5 sec all-out effort, mean power output 902W, 30s
Additional interesting finding is AMPK can be regulated by the Ca2þ passive recovery), speed endurance (6  20 s all-out effort, mean power
sensitive protein calcium/calmodulin-dependent kinase kinase β output 669W, 120s passive recovery) and continuous, modest (50 min at
(CaMKKβ).64,65 Beside AMPK, PGC-1α gene expression is also subjected 70% VO _ 2max, mean power output 218W) exercise protocol with the
to Ca2þ dependent signaling.61,66 participation of 12 young trained men.80 All exercise protocol-induced
AMPK(Thr172), ACC(Ser79) and p38 MAPK(Thr180) phosphorylation
Mitochondrial biogenesis showed no significant differences between groups. NRF2, MFN2 and
TFAM mRNA levels increased only in the speed endurance and contin-
It has been shown that AMPK can activate PGC-1α nuclear coac- uous group.
tivator, which is referred to as the master regulator protein of mito- Another crucial molecule in bioenergetics and metabolism is NADþ
chondrial biogenesis.67 PGC-1α plays an integrative role in governing and also its reduced form NADH. NADH is produced predominantly from
mitochondrial biogenesis. In fact, it has a pivotal role in establishing the NADþ in the citric acid cycle as a product of oxidation. The carbon source
connection between cell energy sensing mechanisms68 and physiological (acetyl-CoA) derived from sugars, fatty- and amino acids, fuels the re-
signal conduction to nuclear transcription factors. The pinnacle aspect of action cycle and provides NADH as a proton donor for the electron
PGC-1α related molecular pathways is its sensitivity to many intracellular transport chain, to run the terminal oxidation. The NADþ/NADH ratio
stimuli and by co-activating NRF1, NRF2, PPARγ, ERR, YY1,69 it regu- and its individual subcellular levels reflect the overall energy and redox
lates the adaptation of the cellular oxidative metabolism. However, in a status of the cell. Sirtuins are NAD þ dependent enzymes81 and SIRT1 has
rather provocative review Islame et al. questioned the dominance of been shown to deacetylate PGC-1α82 and control glucose homeostasis.83
PGC-1α in coordination with mitochondrial biogenesis.70 The authors During high intensity exercise NADH is produced in glycolysis and
main argument is that the sequential regulation of PGC-1α, NRF1/2, transported to the mitochondria for oxidation or, with pyruvate, it is
TFAM, mitochondrial proteins does not always follow this temporal processed by lactate dehydrogenase (LDH) to NADþ and lactate. There is
pattern. They propose that in humans, it is unlikely to have only one a difference of opinion and research rationale, in studies about which
“master regulator” and they point to the possible regulatory role of direction cellular NADþ/NADH changes with exercise volume and in-
PPAR-β, p53 and LRPPRC or LRP130. On the other hand, PGC-1α mRNA tensity.84 The calculated cytosolic NADþ/NADH ratio is reduced85 after
transcription increases with exercise intensity and the PGC-1α mRNA 60 min of cycling exercise at ~62%VO _ 2max or after The Wingate Test.86
abundance is associated with the activation of upstream protein ki- But, according to the data of Phillips and colleagues,87 the calculated
nases.71 Only after high intensity exercise was induced did the down- ratio was reduced after 15 min, then subsequently, returned to resting
stream activating transcription factor-2 (ATF-2) phosphorylation levels by 90 min of exercise at 59%VO _ 2max. Others found NAD þ
increase. _
reduction after 70% and 100% of the VO2max88 with no differences be-
Another level of complexity – which may contribute to the above- _ 2max values. Conversely, others have found
tween 70% and 100% VO
mentioned issue - comes from the promoter structure of the PGC-1α
different trajectories.89 To analyze the exercise-hypoxia- related meta-
(PPARGC1A) gene. Alternative promoter usage coupled with alternative
bolic shift in its complexity in silico methods90 were applied but results
splicing gives rise to eight PGC-1α isoforms ranging from 257 to 797
are still elusive due to the dynamic nature of redox homeostasis and
amino acids in human samples.72 One prominent transcript from the
experimental verifications have yet to be published. Since cytosolic
PGC1α gene alternative promoter is marked as PGC1α4, also known as
NADþ is estimated to be ~540 times higher in skeletal muscle then
NT-PGC1α-b (Martinez-Redondo, Pettersson, & Ruas, 2015). This iso-
NADH it is more likely that NADH is the major player in setting the
form, together with other NT (lacking the N-terminal RS and RR do-
cellular NAD þ /NADH ratio. For a thorough overview the reader is
mains) transcripts, has been shown to be an inducible form. The recently
directed to the paper of White and Schenk.84
emerged question is how these promoters are utilized and what cellular
A remarkable aspect of the redox equivalent conundrum is the high
or physiological events can trigger promoter shifting.73 PGC1α4 expres-
NAD þ level in the mitochondrial compartment.91 Early studies reported
sion has been associated with resistance training74 and it seems that
a positive relationship between resting skeletal muscle NAD þ content
during endurance exercise the transcription of PGC1α4 and other NT
and type I percentage in skeletal muscle.92 Although transition between
isoforms may be intensity-dependent.73,75 To our knowledge, there is no
cytoplasmic, nuclear and mitochondrial metabolite pools are not
comparative study between isoenergetic HIIT and MICT investigating the
barrier-free, the proposed HIIT-induced oxidative shift of type II muscle
differential promoter usage of the PPARGC1A gene.
fibers and its ability to induce mitochondrial biogenesis may give some
It has been reported that 3 h after a single bout of SIT exercise, con-
clue as to why HIIT, with a huge energy demand, activates SIRT142,93
sisting of four all-out cycling sessions, AMPK α1 and α2 were phos-
(Fig. 2).
phorylated in vastus lateralis muscle.76 Moreover, phosphorylation of
ACC and p38 MAPK was also increased compared to the resting state. At
the same time PGC-1α mRNA expression was increased approximately Oxidative stress and the antioxidant system
twofold with no measurable change in the protein level. Similarly, Little
et al. found significant increases in PGC-1α, citrate synthase, COX II and Exercise-derived oxidative challenge makes a major contribution to
COX IV mRNA after 3 3 h of HIIT exercise.77 Nuclear abundance of exercise adaptation and reactive oxygen and nitrogen species are
PGC-1α protein was unchanged immediately after exercise but did in- important signaling molecules in skeletal muscle.94,95 The most promi-
crease 3 h later. PGC-1α protein content in whole muscle lysates nent reactive oxygen sources (and nitrogen) in skeletal muscle are
remained steady immediately after and 3 h after exercise but increased nicotinamide adenine dinucleotide phosphate oxidases (NOX),96
after 24 h of recovery. In a recent study it has been shown that 18 sessions uncoupled nitric oxide synthases (NOS),97 mitochondrial respiratory
of HIIT (10  60-sec cycling intervals at ~90% HRmax, interspersed by enzymes98 and xanthine oxidase (XO).99 Increased exercise intensity
60-sec active recovery at 50 W) increased cytochrome oxidase IV levels in results in enhanced metabolic demand and generation of ROS.9 HIIT
both type I and II muscle fibers.40 HIIT training elevated SIRT3 and training of fourteen bouts of 20-sec swimming exercise induced H2O2

28
F. Torma et al. Sports Medicine and Health Science 1 (2019) 24–32

Fig. 2. Molecular adaptive response of skeletal muscle to HIIT. The suggested molecular pathways by which HIIT results in complex adaptive response to
skeletal muscle.

production in a rat model in permeabilized fibers of tibialis anterior and Conclusion

gastrocnemius but not in soleus muscle.100 We have reported a liner
relationship between blood lactate levels and activity of XO after a single HIIT-associated increases in cellular metabolism of skeletal muscle
bout of exhausted running on the treadmill in a murine model.101 Indeed, results in fiber specific responses, since Type II fibers with higher
it was also reported that high intensity exercise is associated with excess recruital thresholds are fully active during HIIT. The HIIT-induced
production of ROS.102 Conversely, in diseases where inactive lifestyle enhanced mitochondrial biogenesis is mediated by the AMPK, PGC-1α,
plays an important role, tissue specific and systemic ROS elevation is SIRT1 and ROS pathway as well as by the modulation of Ca2þ homeo-
commonly detected.103,104 However, ROS production seems to be stasis. The fact that the time saving HIIT associated adaptive response at
important in insulin regulated glucose uptake105,106 and GLUT4 trans- least comparable or superior to MICT associated adaptation, guarantees
location.107,108 In vitro studies found that H2O2 treatment increased further increases of HIIT in the preparation of elite sport and health
PGC-1α promoter activity and mRNA expression in C2C12 cells.109 promotion physical activities.
Interestingly, HIIT is found to positively regulate GLUT4 protein con-
tent.42,110 ROS are important players in exercise-associated adapta- Conflict of interest
tion,111 since they modulate the Ca channels and force generation,112
mitochondrial biogenesis,102 and housekeeping processes.113 Interest- The authors have no conflict of interest to report.
ingly, the excess of ROS generation during exercise can be partly
controlled by PGC-1α, since activation of this co-activator activates the
Submission statement
expression of antioxidant genes such as SOD2 and GPX.114 Indeed, HIIT
(5  4-min 75% of Wmax), SIT (4  30 s all-out), MICT (30 min at 50% of
The manuscript has not been published and is not under consider-
Wmax) both high intensity exercise, increased peak plasma hydrogen
ation for publication elsewhere.
peroxide values during exercise.115 But, significant elevation of peak
catalase activity was measured only in the SIT group (with a main effect
of time in catalase and SOD activity). It is important to note, that Each Author's contributions
exercise-associated increases in ROS generation very rarely, if ever, can
reach the level which could have a negative effect on health. Mounting FT, ZG, MJ, MT, TM and ZR contributed by searching and discussions
data support the health promoting effects of regular exercise, regardless on the relevant literature. ZR drafted the final version of the paper, but all
of the intensity of exercise. authors were involved in the correction of the original paper. FT and ZG
designed and drew the figures.

29
F. Torma et al. Sports Medicine and Health Science 1 (2019) 24–32

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