medicina

Review
Irisin as a Multifunctional Protein: Implications for
Health and Certain Diseases
Paulina Korta 1 , Ewa Pocheć 1, * and Agnieszka Mazur-Biały 2
1 Department of Glycoconjugate Biochemistry, Institute of Zoology and Biomedical Research,
Faculty of Biology, Jagiellonian University, Gronostajowa 9, 30-387 Krakow, Poland
2 Department of Ergonomics and Exercise Physiology, Faculty of Health Sciences, Jagiellonian University,
Medical College, Grzegorzecka 20, 31-531 Krakow, Poland
* Correspondence: [email protected]; Tel.: +48-12-664-64-67




Received: 29 June 2019; Accepted: 12 August 2019; Published: 15 August 2019


Abstract: Sedentary life style is considered to be an independent risk factor for many disorders,
including development of type 2 diabetes, obesity, immune dysfunction, asthma, and neurological or
coronary heart disease. Irisin is released from myocytes during physical activity, and acts as a link
between muscles and other tissues and organs. This myokine is produced as a result of proteolytic
cleavage of FNDC5 protein present in the membrane of myocytes. Secretion of irisin is regulated
by N-linked oligosaccharides attached to the protein molecule. The two N-glycan molecules, which
constitute a significant part of the irisin glycoprotein, regulate the browning of adipocytes, which is the
most important function of irisin. A receptor specific for irisin has still not been discovered. In some
tissues irisin probably acts via integrins, which are widely expressed transmembrane receptors. Many
studies have confirmed the multifunctional role of irisin and the beneficial effects of this molecule on
body homeostasis. Irisin reduces systemic inflammation, maintains the balance between resorption
and bone formation, and modulates metabolic processes and the functioning of the nervous system.
It suppresses the expression and release of pro-inflammatory cytokines in obese individuals and
attenuates inflammation in adipose tissue. The impact of irisin on cancer cell proliferation, migration,
and invasion has also been demonstrated in numerous studies, which proves its role in carcinogenesis.
Owing to these pleiotropic and beneficial properties, irisin may be a potential option to prevent and
treat civilization-related diseases which are, nowadays, considered to be the major health problems in
Western societies.

Keywords: irisin; FNDC5; N-glycosylation; physical activity; obesity; inflammation; cancers

1. Introduction
Skeletal muscle is the largest organ in the human body [1]. During or immediately after physical
exercise myocytes secrete molecules called myokines, mainly chemokines and cytokines. Myokines
regulate a variety of metabolic processes in various tissues and organs, such as liver, bones, brain, or fat
tissue through endocrine, paracrine, or autocrine signaling pathways. The major myokines include
interleukin 6 (IL-6), monocyte chemotactic protein 1 (MCP1), insulin-like growth factor-1 (IGF-1), and
myostatin [2,3]. In 2012, Boström et al. reported the discovery of a new molecule that is secreted by
myocytes. That molecule was able to induce changes in adipose tissue and activate thermogenesis [4].
Moreover, that molecule was proposed to act as a link between the muscles and other tissues of the
body; the newly discovered protein has been called “irisin,” derived from the name of the Greek
goddess Iris [5]. Since then, irisin has been the subject of extensive research, which enabled the gaining
of insight into its pleiotropic properties.

Medicina 2019, 55, 485; doi:10.3390/medicina55080485 www.mdpi.com/journal/medicina

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2. Structure and N-Glycosylation of Irisin

2. Structure and N-Glycosylation of Irisin
Irisin is a fragment of a cell membrane protein called fibronectin type III domain-containing
proteinIrisin is a fragment of a cell
5 (FNDC5/FRCP2/PeP) [4].membrane
The FNDC5 protein
proteincalled fibronectin
is composed type
of 209 III domain-containing
amino acid (aa) residues.
protein 5 (FNDC5/FRCP2/PeP) [4]. The FNDC5 protein is composed
It contains an N-terminal signal sequence built of 29 aa, a fibronectin type III domain of 209 amino acidwith
(aa) 94
residues.
aa, an
It contains an N-terminal signal sequence built of 29 aa, a fibronectin type
unidentified region consisting of 28 aa, a transmembrane domain having 19 aa, and a C-terminal III domain with 94 aa, an
unidentified region consisting of 28 aa, a transmembrane domain having
part with 39 aa (Figure 1). The C-terminal fragment of FNDC5 is located in the cytoplasm, while the19 aa, and a C-terminal
part with 39 aa
extracellular (Figure 1). potion
N-terminal The C-terminal fragment of
is proteolytically FNDC5
cleaved toisproduce
located in the cytoplasm,
irisin while the
which is ultimately
extracellular
released intoN-terminal potion
the circulation is proteolytically
[6,7]. Both biochemicalcleaved
andtoX-ray
produce irisin which isstudies
crystallography ultimately
showedreleased
that
into the circulation [6,7]. Both biochemical and X-ray crystallography studies showed
irisin occurs in the form of homodimers, where a β-sheet is created between the units. This structure that irisin occurs
in the form of
is stabilized nothomodimers, where bonds,
only by hydrogen a β-sheet
butisalso
created between the
by interactions units. This
between structure
the side chainsisofstabilized
adjacent
not only by
subunits, inhydrogen
particular,bonds, but also
between by interactions
Arg-75 and Glu-79,between
which intheturn sideprotect
chains theof adjacent
dimer endssubunits,
and
in particular, between
Trp-90/Trp-90 Arg-75
[8]. It was and Glu-79,
previously which in that
established turn protect
irisin isthe dimer ends
composed of and
112 Trp-90/Trp-90
aa. The mass [8]. of
It was previously established that irisin is composed of 112
FNDC5 proteins range from 20 to 32 kDa depending on the number and structure ofaa. The mass of FNDC5 proteins range
from 20 to 32 kDa(glycans)
oligosaccharides depending on the to
attached number and structure
the protein moleculeofduring
oligosaccharides (glycans) attached
the post-translational process toof
the protein molecule
N-glycosylation [4,9]. during the post-translational process of N-glycosylation [4,9].

Figure 1.
Figure 1. FNDC5
FNDC5 structure
structure and
and formation
formation of
of irisin.
irisin. The
The potential N-glycosylation sites
potential N-glycosylation sites are
are marked
marked as
as
black dots.
black dots.Asn,
Asn, asparagine;
asparagine; GlcNAc,
GlcNAc, N-acetylglucosamine;
N-acetylglucosamine; Man, mannose;
Man, mannose; Ser,Thr,
Ser, serine; serine; Thr,
threonine;
threonine;
X, any aminoX, any
acidamino
exceptacid except proline.
proline.

Glycosylation is one of the the most

most common
common post-translational
post-translational modifications of proteins which
occurs in in the lumen of endoplasmic reticulum and
the lumen of endoplasmic reticulum and the Golgi
the Golgi apparatus.
apparatus. Morehalf
More than than half
of all of all
proteins
proteins
are are glycosylated,
glycosylated, mainly cell
mainly cell membrane andmembrane and secreted
secreted proteins. proteins.
The attachment The attachment
of carbohydrates, whichof
carbohydrates,
is which isregulated
a multi-stage process a multi-stage processofregulated
by hundreds enzymes,by hundreds
leads of heterogeneity
to a great enzymes, leads to aglycan
in the great
heterogeneity
structures. in the glycan affect
Oligosaccharides structures. Oligosaccharides
the physicochemical affect the
properties physicochemical
of proteins, properties
are necessary of
to obtain
proteins,
the accurateareconformation
necessary to obtain the accurate
of proteins, provideconformation of proteins,
protection against provide
proteolysis, andprotection
are importantagainst
for
proteolysis,
their and
biological are important
functions for metabolic
in different their biological functions
processes in different
[10]. FNDC5 metabolic processes
is an N-glycosylated protein[10].
and
FNDC5 is
contains an N-glycosylated
oligosaccharides protein
attached to theand containsresidue
asparagine oligosaccharides attached tosequence
in the Asn–X–Ser/Thr the asparagine
(where
residue
X in the acid
is any amino Asn–X–Ser/Thr sequence
except proline), (where X is any amino
via a N-acetylglucosamine residueacid except[11].
(GlcNAc) proline),
Three via
maina
N-acetylglucosamine
groups residue (GlcNAc)
of N-glycan structures are linked[11]. Three main groups
via N-glycosidic bonds to ofAsn:
N-glycan structures are linked via
High-mannose/oligomannose
N-glycosidic
type, complexbonds to Asn:
type and High-mannose/oligomannose
hybrid type. All N-glycans containtype, complex
the same type and hybrid
pentasaccharide type.
core but All
differ
N-glycans
in contain the
the composition same
of the sidepentasaccharide core but
chains. The external differofinhigh-mannose
portions the composition of theconsist
glycans side chains.
of 5–9
The external
mannose (Man) portions of high-mannose
residues. Complex-type glycans consist
oligosaccharides of 5–9 mannose
are characterized (Man) structures
by a more diverse residues.
Complex-type
composed oligosaccharides
of GlcNAc, galactoseare characterized
(Gal), by aand
fucose (Fuc), more diverse
sialic acid structures composed
(SA) residues. of GlcNAc,
The hybrid-type
galactose (Gal), fucose (Fuc), and sialic acid (SA) residues. The hybrid-type constitutes an
Medicina 2019, 55, 485 3 of 14
Medicina 2019, 55, x FOR PEER REVIEW 3 of 14

constitutes
intermediate anclass
intermediate
of N-glycans of N-glycans
class where one arm where one arm
is built is built of
of mannose mannose
residues and residues
the otherandarm
the
other arm resembles the structures of complex-type
resembles the structures of complex-type glycans [12]. glycans [12].
The
The glycosylation
glycosylation processprocess of of FNDC5/irisin
FNDC5/irisin is is still
still poorly
poorly characterized.
characterized. The The sequence
sequence of of FNDC5
FNDC5
contains three potential N-glycosylation sites and two of them, Asn-36
contains three potential N-glycosylation sites and two of them, Asn-36 and Asn-81, are occupied by and Asn-81, are occupied
by N-glycans
N-glycans (Figure
(Figure 1). 1). However,
However, theirtheir structure
structure hashasnotnotbeen been determined
determined so sofar.far.
TheThe absence
absence of
of oligosaccharides has a significant effect on the stability of the molecule.
oligosaccharides has a significant effect on the stability of the molecule. It was found that the It was found that the
de-N-glycosylated
de-N-glycosylated FNDC5 FNDC5isismore moresensitive
sensitivetotothe theaction
actionofofprotein
protein synthesis
synthesis inhibitors
inhibitors compared
compared to
to the glycosylated molecule. The removal of one N-glycosylation
the glycosylated molecule. The removal of one N-glycosylation site by site-directed mutagenesissite by site-directed mutagenesis
resulted
resulted in in aa significant
significantreduction
reductionin inthe
thestability
stabilityofofFNDC5,
FNDC5,with witha ahalf-life
half-lifeofof about
about 7 h7 compared
h compared to
to the 12 h characteristic of the fully glycosylated protein. The de-N-glycosylated
the 12 h characteristic of the fully glycosylated protein. The de-N-glycosylated FNDC5 does FNDC5 does not
not
achieve
achieve aa normal
normal spatial
spatialconformation
conformationand andisisnot
notincorporated
incorporatedinto intothe cell
the membrane.
cell membrane. That results
That in
results
ainsignificant decrease in irisin secretion into the blood
a significant decrease in irisin secretion into the blood [11,13]. [11,13].
Irisin
Irisin alsoalsohashas N-glycosylation
two two N-glycosylationsites located
sites atlocated
Asn-7 and at Asn-52
Asn-7 positions
and Asn-52 [14]. Deglycosylation
positions [14].
of
Deglycosylation of irisin lowers its molecular weight to 12 kDa [15] or 15 kDa [14]. Theoraddition
irisin lowers its molecular weight to 12 kDa [15] or 15 kDa [14]. The addition of one two sugarof
chains increases its mass to 22 kDa or 25 kDa, respectively. Both N-glycans
one or two sugar chains increases its mass to 22 kDa or 25 kDa, respectively. Both N-glycans are are probably important to
the primary
probably function to
important of irisin in the browning
the primary function of of adipocytes,
irisin in thewhichbrowningwas evident by an up-regulation
of adipocytes, which was
of mitochondria uncoupling protein-1 (UCP-1) expression
evident by an up-regulation of mitochondria uncoupling protein-1 (UCP-1) expressionand its transcriptional factor peroxisome
and its
proliferator-activated
transcriptional factor receptor
peroxisome γ (PPARγ) coactivator-1α (PGC-1α)
proliferator-activated receptor γ in(PPARγ)
the presence of irisin [14].(PGC-1α)
coactivator-1α Glycans
do notpresence
in the affect theofformation
irisin [14].ofGlycans
irisin dimers
do not(Figure
affect the2) [8].
formation of irisin dimers (Figure 2) [8].

Figure 2.2. The

Figure Therole
roleof of N-glycans
N-glycans attached
attached to FNDC5
to FNDC5 andproteins.
and irisin irisin proteins. Schematic
Schematic structurestructure of
of N-glycan:
N-glycan: Red squares, N-acetylglucosamine; blue circles, mannose; yellow circles, galactose;
Red squares, N-acetylglucosamine; blue circles, mannose; yellow circles, galactose; green triangles, green
triangles,
sialic acid;sialic
brown acid; brownfucose.
hexagons, hexagons, fucose.
Scheme of anScheme of an artery
artery comes comes
from the from
website the website
SMART Sevier SMART
Medical
Sevier
Art Medical Art (https://smart.servier.com)
(https://smart.servier.com) published by Les published by Les
Laboratoires Laboratoires Servier.
Servier.

3. Occurrence, Serum
3. Occurrence, Serum Concentration,
Concentration, and
and Possible
Possible receptors
Receptorsof
ofIrisin
Irisin
Irisin
Irisin isisananadipomyokine
adipomyokine secreted mainly
secreted by skeletal
mainly muscles
by skeletal as well
muscles asaswell
subcutaneous and visceral
as subcutaneous and
adipose
visceral adipose tissues. Immunohistochemical studies showed that smaller amounts of produced
tissues. Immunohistochemical studies showed that smaller amounts of irisin are also irisin are
by
alsotestes, liver, pancreas,
produced by testes, brain,
liver, spleen,
pancreas,heart, andspleen,
brain, stomach [16,17].
heart, andBoström
stomachet[16,17].
al. demonstrated
Boström etthat al.
the level of irisin increases in the blood after physical exercise. They observed a 65%
demonstrated that the level of irisin increases in the blood after physical exercise. They observed increase in irisin
a
concentration
65% increase ininirisinmiceconcentration
running regularly
in miceforrunning
3 weeks. An increase
regularly for 3 in the level
weeks. of this adipomyokine
An increase in the level of
by
thistwo times was also
adipomyokine by found in thewas
two times blood
alsooffound
healthy
inpeople afterof10healthy
the blood weeks of supervised
people training
after 10 weeks[4].of
supervised training [4]. Irisin concentration was found to be higher in active rather than in sedentary
subjects (p = 0.006), and its level also depends on the activities performed at residential place,
Medicina 2019, 55, 485 4 of 14

Irisin concentration was found to be higher in active rather than in sedentary subjects (p = 0.006),
and its level also depends on the activities performed at residential place, because its levels were
found to be significantly higher in the serum of rural individuals compared to urban inhabitants (p
< 0.0001) [18]. Furthermore, physical exercise is known to increase the level of irisin in people with
metabolic disorders [19]. However, the impact of exercise on irisin concentration in the blood is not
clear. People who train regularly usually show a lower irisin serum level [20]. It is suspected that
the type of physical activity plays a role, because irisin upregulation was noted after high-intensity
exercise [21] and after resistance training, but not after endurance exercise [22]. Whole-body vibration
training also contributes to the elevation of irisin concentration [23]. In addition to physical exercise,
diet, and hormonal regulation also affect the irisin levels [24]. The pathological conditions associated
with different diseases significantly influence the release of irisin into the blood circulation. Lower irisin
concentrations were observed in obese individuals and patients suffering from type 2 diabetes [25],
chronic renal failure [26], and prolonged hypothyroidism [27]. The concentration of irisin was found
to be about 3.6 ng/mL in the blood of sedentary people and rises to 4.3 ng/mL in active subjects [15].
Till date, no specific receptor for irisin has been identified. The results of recent studies have shown
that in some tissues irisin exerts its action via binding to integrins, especially the members of αv integrin
family [28]. Integrins are widely expressed transmembrane receptors that bind extracellular matrix
ligands (cell–matrix interactions), membrane proteins of neighboring cells (intercellular interactions),
and recognize soluble ligands [29]. They are responsible for the adhesion, migration, and aggregation
of cells [30]. Stable noncovalent interactions between 18 α-subunits and eight β-subunits produce
24 functionally different integrin heterodimers [29]. Kim et al. demonstrated the binding of irisin to
several integrins present in the fat cells and osteocytes, including α1β1, and with the highest affinity,
to αV β5 integrin. The treatment of osteocytes with irisin significantly increased the phosphorylation
level of focal adhesion kinase (FAK), the major intracellular signal molecule responsible for integrin
signaling. The use of RGD peptide which binds to αvβ5 and acts as an integrin inhibitor, suppressed
any signaling response triggered by irisin. The authors suggest that the heterodimers belonging to the
αv integrin family are probably the main receptors for irisin in all the tissues [28].

4. Pleiotropic Activity of Irisin

4.1. Irisin and Adipose Tissue

One of the most important functions of irisin is the capability to induce changes in adipose tissue.
Based on the structure and function of fat cells, white/yellow (WAT) and brown (BAT) adipose tissues
were distinguished. WAT mainly consists of mature white adipocytes with a peripherally located
nucleus and a single big lipid drop. WAT functions by accumulating excess energy in the form of
triglycerides, protects organs against mechanical damage, and releases adipokines which regulate
various biological processes, including inflammatory reactions [31–33]. Thermogenesis takes place in
BAT and hence is important to maintain the body temperature. BAT is morphologically different from
WAT because it contains many small lipid drops, a centrally located nucleus and a large number of
mitochondria. The lipids present in BAT are primarily used for oxidative phosphorylation and heat
generation. UPC-1, also named thermogenin, is expressed in the mitochondrial membrane of BAT, and
plays a crucial role in these processes [33–35]. During physical exercise, PGC-1α is activated in the
skeletal muscles and regulates the transcription of FNDC5 protein. An increase in PGC-1α level is
accompanied by an elevation of mitochondrial biogenesis. PGC-1α regulates gluconeogenesis and
affects the biosynthesis of heme. In BAT, PGC-1α together with irisin, modulates the expression of
UCP-1 and thermogenesis. As a result, the energy consumption increases and the metabolism of lipids
and glucose are driven [36–38]. Zhang et al. showed that irisin also affects the functioning of WAT,
and the effects of its actions depend on the degree of cell differentiation. The effects of irisin were
evaluated on mature adipocytes and undifferentiated preadipocytes in in vitro studies. Irisin increased
the expression of UCP-1 in mature fat cells leading to the conversion of the phenotype from WAT to
Medicina 2019, 55, 485 5 of 14

BAT by a process named browning. That results in the formation of a third type of adipose tissue called
beige adipocyte tissue. The process of browning was regulated by irisin-induced phosphorylation
of mitogen-activated protein kinases (MAPKs), such as ERK and p38 protein. The treatment of WAT
preadipocytes with irisin did not result in their browning, but inhibited adipogenesis and effectively
reduced the formation of new adipocytes [14,39]. The study of Pérez-Sotelo et al. on the C3H10T1/2
mesenchymal stem cell line confirmed that the lack of FNDC5 expression and secretion of irisin
intensifies adipogenesis and reduces thermogenesis [40]. In addition, irisin enhances sensitivity to
insulin by increasing glycogenesis and reducing gluconeogenesis [37]. Due to these properties, irisin
may be an option to prevent and treat obesity and diabetes.

4.2. Irisin and Nervous System

Physical exercise has a beneficial effect on the functioning of the nervous system, especially on the
hippocampus (the crucial structure for memory and learning), and results in better performance of
cognitive functions [41]. Regular and moderate exercise increased the proliferation and differentiation
of mouse neurons, increased their survival period, and stimulated migration [42]. Irisin plays an
important role in the central nervous system. FNDC5 is expressed in Purkinje cells of the cerebellum [43]
and in rodent hippocampus [44]. Moreover, the increase in FNDC5 expression correlates positively
with the expression of brain-derived neurotrophic factor (BDNF), one of the most important signaling
molecules for synaptic plasticity and neurogenesis in the hippocampus [44,45]. A lack of FNDC5
expression suppressed the differentiation of mouse embryonic stem cells into neurons and interfered
with the maturation of astrocytes [46]. Moon et al. has also shown that irisin may regulate hippocampal
neurogenesis in mice. Pharmacological doses of irisin (50–100 nM) increased the proliferation of mouse
H19-7 hippocampal neuronal cells [47]. Irisin reduced oxidative stress-induced neuronal damage by
inhibiting the secretion of proinflammatory cytokines, such as tumor necrosis factor α (TNFα) and IL-6,
by the Akt/ERK1/2 signaling pathway in the mouse model of cerebral ischemia (MCAO) [48]. A positive
correlation between serum irisin and BDNF levels, and cognition and episodic memory was observed
after 10 weeks of physical training in adult volunteers at risk of dementia [49]. Exercises improve the
mood, and more recently they were also considered to act as antidepressants. In addition to serotonin,
irisin may also contribute to this effect, because decreased levels of this myokine is associated with
mood swings. This effect of irisin is most likely associated with the activation of the PGC-1α/BDNF
pathway [44,50]. Recent research showed that the levels of FNDC5/irisin in cerebrospinal fluid and
hippocampi were reduced in patients with Alzheimer’s disease and the mouse model of this chronic
neurodegenerative disease. The knockdown of FNDC5/irisin in mice brain cells impaired long-term
potentiation and memory in the hippocampal region, while the expression of FNDC5/irisin restored
hippocampal synaptic plasticity and memory in that animal model. Those recent findings revealed
that irisin activity is important to synapse function and memory in a mouse model of Alzheimer’s
disease and confirmed irisin-mediated beneficial effects of exercises on proper functioning of nervous
system [51].

4.3. Irisin and Bones

Physical exercise is quite a strong incentive for stimulating bone formation. It has a beneficial effect
on bone mineral density (BMD), increases the content of minerals, and reduces the risk of fractures, and
hence regular exercise can prevent bone loss associated with the aging processes [52,53]. Colaianni et al.
proved that irisin acts as a link between skeletal muscles and bones. This adipomyokine was found to
exert a beneficial effect on cortical bone development in young mice treated with recombinant irisin for
four weeks at a dose of 100 µg/kg/week. Irisin significantly increased the mass and strength of the
cortical bones and positively modified their geometry by reducing the secretion of osteoblast inhibitors
and causing an activation of activating transcription factor 4 (Atf4), and runt-related transcription
factor 2 (Runx2), which consequently resulted in the expression of bone-specific genes; such as Osx
(encoded an osterix) and Col1a1 (encoded a collagen type I α1), and an increased activity of osteogenic
Medicina 2019, 55, 485 6 of 14

cells [54]. It has also been shown in an in vitro study that irisin enhances the osteoblast differentiation
process [55] through the activation of the aerobic glycolysis pathway [56]. It was suggested that the
MAPK signaling pathway plays a key role in irisin-induced osteogenesis [54,55].
The results of clinical trials also indicate the positive effect of irisin on bone formation. Serbest et al.
measured serum irisin concentration in patients with lower limb bone fractures who underwent an
operation of bone fixation with closed intramedullary nailing. Irisin levels were determined on the day
before surgery, also 1, 15, and 60 days after operation. A two-fold increase in serum concentration was
observed 2 months after the operation compared to the concentration at other time points. The enhanced
secretion of this myokine during bone healing process indicates its anabolic effects [57]. This result
seems to be consistent with that reported by Anastasilakis et al. who showed that osteoporotic fractures
in postmenopausal women have been associated with lower concentrations of irisin [58]. Irisin levels
were lower in female athletes without menstruation in comparison to eumenorrheic athletes and
non-athletes. In all athletes a positive correlation between irisin concentration and volumetric BMD and
bone stiffness was observed [59]. The study by Palermo et al. confirmed the reduction of serum irisin
concentration in donors with previous osteoporotic fractures. However, they reported no relationship
between irisin levels and BMD [60].
Osteocytes in mature bone are found to be sensitive for irisin regulation. As mentioned in
Section 3, this myokine acts on bone via integrin receptors, especially the members of αv integrin
family. The physiological concentration of irisin (3–5 ng/mL) reduced the percentage of MLO-Y4
osteocyte-like cells undergoing apoptosis induced by hydrogen peroxide (H2 O2 ). Besides showing
the protective properties, irisin increased the expression of sclerostin (a specific glycoprotein, which
acts as an inhibitor of bone formation and is responsible for bone remodeling) in MLO-Y4 osteocytes
cultured in vitro and in an in vivo mouse model [28]. The stimulating effect of irisin on sclerostin
expression observed by Kim et al. (2018), was opposite to the previous analysis of human serum
samples using enzyme-linked immunosorbent assays (ELISA) which showed an inverse correlation
between sclerostin and irisin levels [61].
The extensive research on the role of irisin in bone physiology underlines its important function
in maintaining homeostasis between resorption and bone formation. The effects of irisin on osteocytes
need further clarification.

4.4. Irisin in Inflammation

A lack of physical activity is an independent risk factor for the development of many chronic
diseases, such as type 2 diabetes, obesity, immune dysfunction, asthma, and neurological or coronary
heart disease. Most of those pathologies are associated with persistent, chronic inflammation. Regular
and moderate physical activity positively affects the functioning of the immune system and reduces
systemic inflammation [62]. Skeletal muscles modulate the inflammatory response mainly by the
secretion of myokines [63], including irisin.
Our research demonstrated that regular physical training may alleviate the symptoms
of inflammatory bowel diseases in obese mice, among others, by inhibiting the secretion of
proinflammatory cytokines and increasing the amount of irisin released [64]. In vitro studies on
RAW 264.7 macrophages showed that irisin regulates the activation of immunocompetent cells.
It enhances the activity and proliferation of macrophages, improves their ability of phagocytosis, and
reduces the production of reactive oxygen species (ROS) without affecting cell viability [65]. Irisin
significantly reduces the extensive production of harmful H2 O2 by macrophages. This antioxidative
activity of irisin is a consequence of increased expression of key antioxidative enzymes, including
superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase 9 (Cat-9) [66]. The high
concentrations of irisin (50 nM, 100 nM) reduced the number of both early and late apoptotic cells,
attenuated the surface expression of Toll-like receptors 4 (TLR4), and diminished the amount of
MyD88 adapter protein in lipopolysaccharide (LPS)-stimulated macrophages. The suppression of
MAPK cascade signaling pathways and a consequential reduction in the levels of NF-κB nuclear
Medicina 2019, 55, 485 7 of 14

transcription factor, c-Jun N-terminal kinases (JNK), and extracellular signal-regulated kinase (ERK)
phosphorylation were also noticed. That led to a decrease in the release of key pro-inflammatory
cytokines, such as IL-1β, IL-6, TNF-α, keratinocyte chemoattractant (KC), macrophage chemotactic
protein (MCP)-1, and high mobility group box 1 (HMGB1) protein [67]. The results of these studies
contribute to the understanding of the molecular mechanism underlying the anti-inflammatory effects
caused by physical activity, and confirm the protective effect of irisin in the development of diseases
associated with chronic inflammation.

4.5. Irisin in Obesity

Obesity is a state of excessive fat accumulation and is usually accompanied by insulin resistance,
type 2 diabetes, cardiovascular diseases, and cancers [68]. The changes in the irisin levels were
observed in the serum of obese people. Earlier studies reported the reduced concentration of irisin
in obese individuals [25], while some of the recent studies showed the opposite results wherein the
irisin levels were found to be increased in obese conditions [69,70]. Sahin-Efe et al. suggested a state of
irisin resistance may be noticed during the course of obesity development (similar to leptin resistance)
which could explain the elevated levels of irisin in these subjects [69]. According to Pardo et al.,
an increase of fat mass by 1 kg can lead to a twofold elevation of irisin level [71], while two other studies
demonstrated that weight loss in obese subjects leads to a reduction of serum irisin levels [72,73].
Obesity is often related to the development of middle inflammation [68,74,75], also referred to as
meta-inflammation, which is the state of chronic inflammation induced in non-immunocompetent
tissues, such as muscles, intestines, adipose tissue, or liver, as a consequence of, e.g., the activation of
resident macrophages [76]. Meta-inflammation may result in various metabolic abnormalities [77],
and during the course of obesity, a massive influx of macrophages into fat tissue is observed [74,78].
Our recent study has shown that adipocytes are sensitive to irisin [79]. LPS-activated mature
adipocytes mimic the state of obesity-related inflammation in vitro and when cultured in the presence
of irisin, they produced and released significantly less amounts of pro-inflammatory cytokines, such
as TNF-α, and IL-6. Irisin also decreased MCP-1 expression, thereby reducing chemotactic influx
of macrophages. Furthermore, irisin impaired the expression and release of leptin, an adipokine
associated with pro-inflammatory activation, and upregulated the level of anti-inflammatory cytokine
adiponectin [79]. The elevated concentration of leptin in the serum of obese subjects contributes
to excess food intake and reduced energy expenditure [80], and can result in the development of
leptin resistance [81]. Leptin activity is closely associated with insulin resistance and the development
of metabolic syndrome [82]. Gutierrez-Repiso et al. demonstrated that leptin plays a role in the
expression of FNDC5 in adipose tissue [83]. Adiponectin, an anti-inflammatory adipokine, increases
the sensitivity of the cell to insulin, whose level is found to be decreased in obesity, and may be
associated with the development of insulin resistance [80]. Huo et al. demonstrated that irisin via
heme oxygenase 1 (HO-1)/adiponectin axis improves perivascular adipose tissue (PVAT) function
in diet-induced obese mice and attenuates the anti-contraction effect of PVAT [84]. Irisin improved
endothelial function in obese subjects via activation of AMPK-eNOS pathway [85]. The protective
effect of irisin was manifested by a reduction of TNF-α, an augmentation of adiponectin level, and
an upregulation of lipid and glucose metabolism in mice fed on a high-fat diet [84]. Considering the
positive anti-inflammatory effects of irisin on both adipocytes and macrophages, it seems reasonable to
look for factors that increase sensitivity to irisin.

4.6. Irisin in Carcinogenesis

Irisin is also involved in carcinogenesis, although its role in cancer progression is currently
ambiguous. It was found that higher concentrations of this adipomyokine in women are associated
with a lower risk of breast cancer [86]. Women with primary breast cancer showed higher concentration
of irisin in comparison to patients with spinal metastasis (p = 0.022) [87]. On the other hand, patients
with renal cell cancer have higher levels of serum irisin (p = 0.0001) compared to the controls [88].
Medicina 2019, 55, 485 8 of 14

In vitro studies showed that irisin suppresses cell proliferation, migration, and viability of MCF-7 and
MDA-MB-231 breast cancer malignant cell lines by stimulating caspase activity and inducing apoptosis,
but do not affect MCF-10A normal breast epithelial cells. Moreover, the sensitivity of MDA-MB-231
malignant cells to the cytotoxic antineoplastic antibiotic doxorubicin (Dox) was increased in the
presence of irisin, and it resulted in the higher breast cancer cytotoxicity [89]. Irisin also inhibited the
growth, migration, and invasion of MIA PaCa-2 and Panc03.27 pancreatic cancer cells cultured in vitro
via the AMPK-mTOR pathway signaling, by increased phosphorylation of AMP-activated protein
kinase (AMPKα) and reduction of mammalian target of rapamycin (mTOR) phosphorylation [90].
A similar effect on cell proliferation and motility was also found in the case of U2OS and MG-63
osteosarcoma cell lines. An inversion of IL-6-induced epithelial–mesenchymal transition (EMT) and an
inhibition of STAT3/Snail pathway signaling were observed in the presence of irisin [91]. The protective
effect of irisin was also demonstrated for A549 and NCI-H446 lung cancer cells which exhibited
suppressed EMT and invasion by decreasing the expression of Snail in the PI3K/Akt/Snail pathway [92].
The anti-cancer activity of irisin was not confirmed for human nor mouse colon, esophageal, thyroid
and endometrial cell lines, which exhibited no change in the proliferation and adhesion properties
in the presence of this myokine [93]. While cell proliferation, migration, and invasion of HepG2 and
SMCC7721 hepatocellular cell lines were promoted by irisin, their sensitivity to Dox was reduced
due to the activation of the Akt/PI3K pathway [94]. The above-mentioned discrepancy suggests that
the action and role of irisin in the initiation, promotion, and progression of cancer may be tissue and
cell specific.

5. Conclusions
The health-promoting effects of physical activity have always been known. The discovery of irisin
in 2012 by Boström et al. revealed for the first time the role of this protein in causing the beneficial effects
of exercise at a macromolecular level. Shortly after the discovery of irisin, numerous studies proved
that this myokine is released into the circulation during physical exercise and that it is a multifunctional
protein that regulates the biological functioning of various cells and tissues (Table 1). The activity and
serum concentration of this myokine depends on the physiological and/or pathological state.

Table 1. Irisin activity and functions in selected physiological and pathological conditions.

Physiological/Pathological
Irisin Activity References
Conditions
Adipose (obesity)
—Modulation of UCP-1 expression in mitochondria and
[36–38]
BAT enhancement of thermogenesis
—Increase of energy consumption and metabolism of lipids and
glucose
—Increase of UCP-1 expression, leading to a conversion of the
WAT [14,39]
phenotype to BAT (browning)
—Induction of hippocampal neurogenesis by regulation of
[44,47]
Nervous system BDNF expression
—Reduction of neuronal injury [48]
—Increase of cortical bone mass and strength, enhancement of
[54,55]
osteoblast differentiation process
Bones
—Induction or inhibition of sclerostin expression [28,61]
—Increase in volumetric BMD or no relationship [59,60]
—Enhancement of macrophage activity and proliferation,
[65]
improvement of phagocytosis, and reduction of ROS production
Inflammation
—Increase of the expression of antioxidative enzymes [66]
—Reduction of the release of proinflammatory cytokines [67]
—Suppression of proliferation, migration, and viability of breast,
[89–92]
pancreatic, osteosarcoma and lung cancer cells
Carcinogenesis
—No effect on colon, esophageal, thyroid, and endometrial
[93]
cancer cell progression
—Promotion of proliferation, migration, and invasion of
[94]
hepatocellular cancer cells
Medicina 2019, 55, 485 9 of 14

Most of the functions predicted for irisin require further studies to confirm or verify the previous
results. First of all, the precise mechanism of irisin action has to be determined, from an interaction with
the receptor, through a cascade of the intracellular signal transmission, to the direct effect on target cell
metabolism, or with the release of biologically active factors. A recombinant non-glycosylated form of
irisin has been used in most of the functional studies. As it was mentioned in Section 2, N-glycosylation
of irisin is crucial to its secretion and the browning of adipocytes (Figure 2), and it seems possible that
N-glycans also affect other activities of irisin. Future studies should be aimed at determining other
glycosylation-dependent irisin functions and potential glycosylation changes that occur in pathological
conditions, because numerous studies have demonstrated that glycosylation of different proteins is
subject to change, mainly in carcinogenesis [95,96] and inflammation [97]. The detailed analysis of
irisin glycosylation patterns and the effects of this post-translational modification on irisin actions are
also crucial in the context of prophylactic and therapeutic uses of irisin.

Author Contributions: P.K. prepared the original draft of manuscript and the Figure 1 and Table 1; E.P. reviewed
and edited the manuscript, and drew Figure 2; A.M.-B. wrote Section 4.5 and reviewed the manuscript.
Funding: This review paper was supported by the research projects No. N43/DBS/000047 (Department of
Ergonomics and Exercise Physiology, Faculty of Health Sciences, Jagiellonian University, Medical College) and
K/ZDS/008062 (Department of Glycoconjugate Biochemistry, Faculty of Biology, Jagiellonian University).
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Amengual, J.; García-Carrizo, F.J.; Arreguín, A.; Mušinović, H.; Granados, N.; Palou, A.; Bonet, M.L.; Ribot, J.
Retinoic Acid Increases Fatty Acid Oxidation and Irisin Expression in Skeletal Muscle Cells and Impacts
Irisin In Vivo. Cell. Physiol. Biochem. 2018, 46, 187–202. [CrossRef] [PubMed]
2. Johnson, R.W.; White, J.D.; Walker, E.C.; Martin, T.J.; Sims, N.A. Myokines (muscle-derived cytokines and
chemokines) including ciliary neurotrophic factor (CNTF) inhibit osteoblast differentiation. Bone 2014, 64,
47–56. [CrossRef] [PubMed]
3. Di Raimondo, D.; Miceli, G.; Musiari, G.; Tuttolomondo, A.; Pinto, A. New insights about the putative role of
myokines in the context of cardiac rehabilitation and secondary cardiovascular prevention. Ann. Transl. Med.
2017, 5, 300. [CrossRef] [PubMed]
4. Boström, P.; Wu, J.; Jedrychowski, M.P.; Korde, A.; Ye, L.; Lo, J.C.; Rasbach, K.A.; Boström, E.A.; Choi, J.H.;
Long, J.Z.; et al. A PGC1-α-dependent myokine that drives brown-fat-like development of white fat and
thermogenesis. Nature 2012, 481, 463–468. [CrossRef] [PubMed]
5. Pukajło, K.; Kolackov, K.; Łaczmański, Ł.; Daroszewski, J. Irisin – a new mediator of energy homeostasis.
Postepy Hig. Med. Dosw. 2015, 69, 233–242. [CrossRef] [PubMed]
6. Panati, K.; Narala, V.R.; Narasimha, V.R.; Derangula, M.; Arva Tatireddigari, V.R.R.; Yeguvapalli, S. Expression,
purification and biological characterisation of recombinant human irisin (12.5 kDa). J. Genet. Eng. Biotechnol.
2018, 16, 459–466. [CrossRef]
7. Mahgoub, M.O.; D’Souza, C.; Al Darmaki, R.S.M.H.; Baniyas, M.M.Y.H.; Adeghate, E. An update on the role
of irisin in the regulation of endocrine and metabolic functions. Peptides 2018, 104, 15–23. [CrossRef]
8. Schumacher, M.A.; Chinnam, N.; Ohashi, T.; Shah, R.S.; Erickson, H.P. The structure of irisin reveals a novel
intersubunit β-sheet fibronectin type III (FNIII) dimer: Implications for receptor activation. Biol. Chem. 2013,
288, 33738–33744. [CrossRef]
9. Roca-Rivada, A.; Castelao, C.; Senin, L.L.; Landrove, M.O.; Baltar, J.; Crujeiras, A.B.; Seoane, L.M.;
Casanueva, F.F.; Pardo, M. FNDC5/irisin is not only a myokine but also an adipokine. PLoS ONE 2013, 8,
e60563. [CrossRef]
10. Korta, P.; Pocheć, E. Glycosylation of thyroid-stimulating hormone receptor. Endokrynol. Pol. 2019, 70, 86–100.
[CrossRef]
11. Nie, Y.; Liu, D. N-Glycosylation is required for FDNC5 stabilization and irisin secretion. Biochem. J. 2017,
474, 3167–3177. [CrossRef] [PubMed]
12. Polak, K.; Pocheć, E. Glycoproteins of immune system: Oligosaccharide structure and function of the selected
t cell membrane receptors–Part I. Post. Biol. Kom. 2017, 44, 185–200.
Medicina 2019, 55, 485 10 of 14

13. Tan, N.Y.; Bailey, U.M.; Jamaluddin, M.F.; Mahmud, S.H.; Raman, S.C.; Schulz, B.L. Sequence-based protein
stabilization in the absence of glycosylation. Nat. Commun. 2014, 5, 3099. [CrossRef] [PubMed]
14. Zhang, Y.; Li, R.; Meng, Y.; Li, S.; Donelan, W.; Zhao, Y.; Qi, L.; Zhang, M.; Wang, X.; Cui, T.; et al. Irisin
stimulates browning of white adipocytes through mitogen-activated protein kinase p38 MAP kinase and
ERK MAP kinase signaling. Diabetes 2014, 63, 514–525. [CrossRef] [PubMed]
15. Jedrychowski, M.P.; Wrann, C.D.; Paulo, J.A.; Gerber, K.K.; Szpyt, J.; Robinson, M.M.; Nair, K.S.; Gygi, S.P.;
Spiegelman, B.M. Detection and Quantitation of Circulating Human Irisin by Tandem Mass Spectrometry.
Cell Metab. 2015, 22, 734–740. [CrossRef] [PubMed]
16. Aydin, S.; Kuloglu, T.; Aydin, S.; Kalayci, M.; Yilmaz, M.; Cakmak, T.; Albayrak, S.; Gungor, S.; Colakoglu, N.;
Ozercan, I.H. A comprehensive immunohistochemical examination of the distribution of the fat-burning
protein irisin in biological tissues. Peptides 2014, 61, 130–136. [CrossRef]
17. Martinez Munoz, I.Y.; Camarillo Romero, E.D.S.; Garduno Garcia, J.J. Irisin a Novel Metabolic Biomarker:
Present Knowledge and Future Directions. Int. J. Endocrinol. 2018, 2018, 7816806. [CrossRef]
18. Moreno, M.; Moreno-Navarrete, J.M.; Serrano, M.; Ortega, F.; Delgado, E.; Sanchez-Ragnarsson, C.; Valdés, S.;
Botas, P.; Ricart, W.; Fernández-Real, J.M. Circulating irisin levels are positively associated with metabolic
risk factors in sedentary subjects. PLoS ONE 2015, 10, e0124100. [CrossRef]
19. Huh, J.Y.; Siopi, A.; Mougios, V.; Park, K.H.; Mantzoros, C.S. Irisin in response to exercise in humans with
and without metabolic syndrome. J. Clin. Endocrinol. Metab. 2015, 100, E453–E457. [CrossRef]
20. Qiu, S.; Cai, X.; Sun, Z.; Schumann, U.; Zügel, M.; Steinacker, J.M. Chronic Exercise Training and Circulating
Irisin in Adults: A Meta-Analysis. Sports Med. 2015, 45, 1577–1588. [CrossRef]
21. Tsuchiya, Y.; Ando, D.; Goto, K.; Kiuchi, M.; Yamakita, M.; Koyama, K. High-intensity exercise causes greater
irisin response compared with low-intensity exercise under similar energy consumption. Tohoku J. Exp. Med.
2014, 233, 135–140. [CrossRef] [PubMed]
22. Tsuchiya, Y.; Ando, D.; Takamatsu, K.; Goto, K. Resistance exercise induces a greater irisin response than
endurance exercise. Metabolism 2015, 64, 1042–1050. [CrossRef] [PubMed]
23. Greulich, T.; Nell, C.; Koepke, J.; Fechtel, J.; Franke, M.; Schmeck, B.; Haid, D.; Apelt, S.; Filipovic, S.;
Kenn, K.; et al. Benefits of whole body vibration training in patients hospitalised for COPD exacerbations—A
randomized clinical trial. BMC Pulm. Med. 2014, 14, 60. [CrossRef]
24. Varela-Rodríguez, B.M.; Pena-Bello, L.; Juiz-Valiña, P.; Vidal-Bretal, B.; Cordido, F.; Sangiao-Alvarellos, S.
FNDC5 expression and circulating irisin levels are modified by diet and hormonal conditions in hypothalamus,
adipose tissue and muscle. Sci. Rep. 2016, 6, 29898. [CrossRef] [PubMed]
25. Moreno-Navarrete, J.M.; Ortega, F.; Serrano, M.; Guerra, E.; Pardo, G.; Tinahones, F.; Ricart, W.;
Fernández-Real, J.M. Irisin is expressed and produced by human muscle and adipose tissue in association
with obesity and insulin resistance. J. Clin. Endocrinol. Metab. 2013, 98, E769–778. [CrossRef] [PubMed]
26. Liu, J.J.; Liu, S.; Wong, M.D.; Tan, C.S.; Tavintharan, S.; Sum, C.F.; Lim, S.C. Relationship between circulating
irisin, renal function and body composition in type 2 diabetes. J. Diabetes Complicat. 2014, 28, 208–213.
[CrossRef]
27. Zybek-Kocik, A.; Sawicka-Gutaj, N.; Wrotkowska, E.; Sowiński, J.; Ruchała, M. Time-dependent irisin
concentration changes in patients affected by overt hypothyroidism. Endokrynol. Pol. 2016, 67, 476–480.
[CrossRef]
28. Kim, H.; Wrann, C.D.; Jedrychowski, M.; Vidoni, S.; Kitase, Y.; Nagano, K.; Zhou, C.; Chou, J.; Parkman, V.A.;
Novick, S.J.; et al. Irisin Mediates Effects on Bone and Fat via αV Integrin Receptors. Cell 2018, 175, 1756–1768.
[CrossRef] [PubMed]
29. Takada, Y.; Ye, X.; Simon, S. The integrins. Genome Biol. 2007, 8, 215. [CrossRef]
30. Czyż, M. Regulacja ekspresji integryn. Acta Haematol. Pol. 2000, 31, 17–23.
31. Fenzl, A.; Kiefer, F.W. Brown adipose tissue and thermogenesis. Horm. Mol. Biol. Clin. Investig. 2014, 19,
25–37. [CrossRef] [PubMed]
32. Lee, Y.H.; Mottillo, E.P.; Granneman, J.G. Adipose tissue plasticity from WAT to BAT and in between. Biochim.
Biophys. Acta 2014, 1842, 358–369. [CrossRef] [PubMed]
33. Moonen, M.P.B.; Nascimento, E.B.; Van Marken Lichtenbelt, W.D. Human brown adipose tissue:
Underestimated target in metabolic disease? Biochim. Biophys. Acta Mol. Cell Biol. Lipids 2019, 1864,
104–112. [CrossRef] [PubMed]
Medicina 2019, 55, 485 11 of 14

34. Schulz, T.J.; Tseng, Y.H. Brown adipose tissue: Development, metabolism and beyond. Biochem. J. 2013, 453,
167–178. [CrossRef] [PubMed]
35. Lidell, M.E.; Betz, M.J.; Enerbäck, S. Brown adipose tissue and its therapeutic potential. J. Intern. Med. 2014,
276, 364–377. [CrossRef] [PubMed]
36. Handschin, C.; Spiegelman, B.M. Peroxisome proliferator-activated receptor gamma coactivator 1 coactivators,
energy homeostasis, and metabolism. Endocr. Rev. 2006, 27, 728–735. [CrossRef]
37. Gouveia, M.C.; Vella, J.P.; Cafeo, F.R. Affonso Fonseca, F.L.; Bacci, M.R. Association between irisin and major
chronic diseases: A review. Eur. Rev. Med. Pharmacol. Sci. 2016, 20, 4072–4077. [PubMed]
38. Grygiel-Górniak, B.; Puszczewicz, M. A review on irisin, a new protagonist that mediates
muscle-adipose-bone-neuron connectivity. Eur. Rev. Med. Pharmacol. Sci. 2017, 21, 4687–4693.
39. Zhang, Y.; Xie, C.; Wang, H.; Foss, R.M.; Clare, M.; George, E.V.; Li, S.; Katz, A.; Cheng, H.; Ding, Y.; et al.
Irisin exerts dual effects on browning and adipogenesis of human white adipocytes. Am. J. Physiol. Endocrinol.
Metab. 2016, 311, E530–E541. [CrossRef]
40. Pérez-Sotelo, D.; Roca-Rivada, A.; Baamonde, I.; Baltar, J.; Castro, A.I.; Domínguez, E.; Collado, M.;
Casanueva, F.F.; Pardo, M. Lack of Adipocyte-Fndc5/Irisin Expression and Secretion Reduces Thermogenesis
and Enhances Adipogenesis. Sci. Rep. 2017, 7, 16289. [CrossRef]
41. Wrann, C.D. FNDC5/irisin-Their role in the nervous system and as a mediator for beneficial effects of exercise
on the brain. Brain Plast. 2015, 1, 55–61. [CrossRef] [PubMed]
42. So, J.H.; Huang, C.; Ge, M.; Cai, G.; Zhang, L.; Lu, Y.; Mu, Y. Intense Exercise Promotes Adult Hippocampal
Neurogenesis But Not Spatial Discrimination. Front. Cell. Neurosci. 2017, 11, 13. [CrossRef] [PubMed]
43. Dun, S.L.; Lyu, R.M.; Chen, Y.H.; Chang, J.K.; Luo, J.J.; Dun, N.J. Irisin-immunoreactivity in neural and
non-neural cells of the rodent. Neuroscience 2013, 240, 155–162. [CrossRef] [PubMed]
44. Wrann, C.D.; White, J.P.; Salogiannnis, J.; Laznik-Bogoslavski, D.; Wu, J.; Ma, D.; Lin, J.D.; Greenberg, M.E.;
Spiegelman, B.M. Exercise induces hippocampal BDNF through a PGC-1α/FNDC5 pathway. Cell Metab.
2013, 18, 649–659. [CrossRef] [PubMed]
45. Hassanzadeh, S.; Jameie, S.B.; Mehdizadeh, M.; Soleimani, M.; Namjoo, Z.; Soleimani, M. FNDC5 expression in
Purkinje neurons of adult male rats with acute spinal cord injury following treatment with methylprednisolone.
Neuropeptides 2018, 70, 16–25. [CrossRef] [PubMed]
46. Hashemi, M.S.; Ghaedi, K.; Salamian, A.; Karbalaie, K.; Emadi-Baygi, M.; Tanhaei, S.; Nasr-Esfahani, M.H.;
Baharvand, H. Fndc5 knockdown significantly decreased neural differentiation rate of mouse embryonic
stem cells. Neuroscience 2013, 231, 296–304. [CrossRef] [PubMed]
47. Moon, H.S.; Dincer, F.; Mantzoros, C.S. Pharmacological concentrations of irisin increase cell proliferation
without influencing markers of neurite outgrowth and synaptogenesis in mouse H19-7 hippocampal cell
lines. Metabolism 2013, 62, 1131–1136. [CrossRef]
48. Li, D.J.; Li, Y.H.; Yuan, H.B.; Qu, L.F.; Wang, P. The novel exercise-induced hormone irisin protects
against neuronal injury via activation of the Akt and ERK1/2 signaling pathways and contributes to the
neuroprotection of physical exercise in cerebral ischemia. Metabolism 2017, 68, 31–42. [CrossRef]
49. Küster, O.C.; Laptinskaya, D.; Fissler, P.; Schnack, C.; Zügel, M.; Nold, V.; Thurm, F.; Pleiner, S.;
Karabatsiakis, A.; von Einem, B.; et al. Novel Blood-Based Biomarkers of Cognition, Stress, and Physical or
Cognitive Training in Older Adults at Risk of Dementia: Preliminary Evidence for a Role of BDNF, Irisin,
and the Kynurenine Pathway. J. Alzheimers Dis. 2017, 59, 1097–1111. [CrossRef]
50. de Oliveira Bristot, V.J.; de Bem Alves, A.C.; Cardoso, L.R.; da Luz Scheffer, D.; Aguiar, A.S., Jr. The Role of
PGC-1α/UCP2 Signaling in the Beneficial Effects of Physical Exercise on the Brain. Front. Neurosci. 2019, 13,
292. [CrossRef]
51. Lourenco, M.V.; Frozza, R.L.; de Freitas, G.B.; Zhang, H.; Kincheski, G.C.; Ribeiro, F.C.; Gonçalves, R.A.;
Clarke, J.R.; Beckman, D.; Staniszewski, A.; et al. Exercise-linked FNDC5/irisin rescues synaptic plasticity
and memory defects in Alzheimer’s models. Nat. Med. 2019, 25, 165–175. [CrossRef] [PubMed]
52. Andreoli, A.; Celi, M.; Volpe, S.L.; Sorge, R.; Tarantino, U. Long-term effect of exercise on bone mineral
density and body composition in post-menopausal ex-elite athletes: A retrospective study. Eur. J. Clin. Nutr.
2012, 66, 69–74. [CrossRef] [PubMed]
53. Polyzos, S.A.; Anastasilakis, A.D.; Efstathiadou, Z.A.; Makras, P.; Perakakis, N.; Kountouras, J.; Mantzoros, C.S.
Irisin in metabolic diseases. Endocrine 2018, 59, 260–274. [CrossRef] [PubMed]
Medicina 2019, 55, 485 12 of 14

54. Colaianni, G.; Cuscito, C.; Mongelli, T.; Pignataro, P.; Buccoliero, C.; Liu, P.; Lu, P.; Sartini, L.; Di Comite, M.;
Mori, G.; et al. The myokine irisin increases cortical bone mass. Proc. Natl. Acad. Sci. USA 2015, 112,
12157–12162. [CrossRef] [PubMed]
55. Qiao, X.; Nie, Y.; Ma, Y.; Chen, Y.; Cheng, R.; Yin, W.; Hu, Y.; Xu, W.; Xu, L. Irisin promotes osteoblast
proliferation and differentiation via activating the MAP kinase signaling pathways. Sci. Rep. 2016, 6, 18732.
[CrossRef] [PubMed]
56. Zhang, D.; Bae, C.; Lee, J.; Lee, J.; Jin, Z.; Kang, M.; Cho, Y.S.; Kim, J.H.; Lee, W.; Lim, S.K. The bone anabolic
effects of irisin are through preferential stimulation of aerobic glycolysis. Bone 2018, 114, 150–160. [CrossRef]
[PubMed]
57. Serbest, S.; Tiftikçi, U.; Tosun, H.B.; Kısa, Ü. The Irisin Hormone Profile and Expression in Human Bone
Tissue in the Bone Healing Process in Patients. Med. Sci. Monit. 2017, 23, 4278–4283. [CrossRef]
58. Anastasilakis, A.D.; Polyzos, S.A.; Makras, P.; Gkiomisi, A.; Bisbinas, I.; Katsarou, A.; Filippaios, A.;
Mantzoros, C.S. Circulating irisin is associated with osteoporotic fractures in postmenopausal women with
low bone mass but is not affected by either teriparatide or denosumab treatment for 3 months. Osteoporos.
Int. 2014, 25, 1633–1642. [CrossRef]
59. Singhal, V.; Lawson, E.A.; Ackerman, K.E.; Fazeli, P.K.; Clarke, H.; Lee, H.; Eddy, K.; Marengi, D.A.;
Derrico, N.P.; Bouxsein, M.L.; et al. Irisin levels are lower in young amenorrheic athletes compared with
eumenorrheic athletes and non-athletes and are associated with bone density and strength estimates. PLoS
ONE 2014, 9, e100218. [CrossRef]
60. Palermo, A.; Strollo, R.; Maddaloni, E.; Tuccinardi, D.; D’Onofrio, L.; Briganti, S.I.; Defeudis, G.; De
Pascalis, M.; Lazzaro, M.C.; Colleluori, G.; et al. Irisin is associated with osteoporotic fractures independently
of bone mineral density, body composition or daily physical activity. Clin. Endocrinol. 2015, 82, 615–619.
[CrossRef]
61. Klangjareonchai, T.; Nimitphong, H.; Saetung, S.; Bhirommuang, N.; Samittarucksa, R.; Chanprasertyothin, S.;
Sudatip, R.; Ongphiphadhanakul, B. Circulating sclerostin and irisin are related and interact with gender to
influence adiposity in adults with prediabetes. Int. J. Endocrinol. 2014, 2014, 261545. [CrossRef] [PubMed]
62. Handschin, C.; Spiegelman, B.M. The role of exercise and PGC1alpha in inflammation and chronic disease.
Nature 2008, 454, 463–469. [CrossRef] [PubMed]
63. Díaz, B.B.; González, D.A.; Gannar, F.; Pérez, M.C.R.; De León, A.C. Myokines, physical activity, insulin
resistance and autoimmune diseases. Immunol. Lett. 2018, 203, 1–5. [CrossRef] [PubMed]
64. Mazur-Bialy, A.I.; Bilski, J.; Wojcik, D.; Brzozowski, B.; Surmiak, M.; Hubalewska-Mazgaj, M.; Chmura, A.;
Magierowski, M.; Magierowska, K.; Mach, T.; et al. Beneficial Effect of Voluntary Exercise on Experimental
Colitis in Mice Fed a High-Fat Diet: The Role of Irisin, Adiponectin and Proinflammatory Biomarkers.
Nutrients 2017, 9, 410. [CrossRef] [PubMed]
65. Mazur-Bialy, A.I. Irisin acts as a regulator of macrophages host defense. Life Sci. 2017, 176, 21–25. [CrossRef]
[PubMed]
66. Mazur-Bialy, A.I.; Kozlowska, K.; Pochec, E.; Bilski, J.; Brzozowski, T. Myokine irisin-induced protection
against oxidative stress in vitro. Involvement of heme oxygenase-1 and antioxidazing enzymes superoxide
dismutase-2 and glutathione peroxidase. J. Physiol. Pharmacol. 2018, 69, 117–125. [PubMed]
67. Mazur-Bialy, A.I.; Pocheć, E.; Zarawski, M. Anti-Inflammatory Properties of Irisin, Mediator of Physical
Activity, Are Connected with TLR4/MyD88 Signaling Pathway Activation. Int. J. Mol. Sci. 2017, 18, 701.
[CrossRef]
68. Meilianna, A.; Dewi, N.M.; Wijaya, A. Adipose tissue, inflammation (Meta-inflammation) and Obesity
management. Indones. Biomed. J. 2015, 7, 129–146. [CrossRef]
69. Sahin-Efe, A.; Upadhyay, J.; Ko, B.J.; Dincer, F.; Park, K.H.; Migdal, A.; Vokonas, P.; Mantzoros, C. Irisin
and leptin concentrations in relation to obesity, and developing type 2 diabetes: A cross sectional and a
prospective case-control study nested in the Normative Aging Study. Metabolism 2018, 79, 24–32. [CrossRef]
70. De Meneck, F.; de Souza, L.V.; Oliveira, V.; do Franco, M.C. High irisin levels in overweight/obese children
and its positive correlation with metabolic profile, blood pressure, and endothelial progenitor cells. Nutr.
Metab. Cardiovasc. Dis. 2018, 28, 756–764. [CrossRef]
71. Pardo, M.; Crujeiras, A.B.; Amil, M.; Aguera, Z.; Jiménez-Murcia, S.; Baños, R.; Botella, C.; de la Torre, R.;
Estivill, X.; Fagundo, A.B.; et al. Association of irisin with fat mass, resting energy expenditure, and daily
activity in conditions of extreme body mass index. Int. J. Endocrinol. 2014, 2014, 857270. [CrossRef] [PubMed]
Medicina 2019, 55, 485 13 of 14

72. Crujeiras, A.B.; Zulet, M.A.; Lopez-Legarrea, P.; de la Iglesia, R.; Pardo, M.; Carreira, M.C.; Martinez, J.A.;
Casanueva, F.F. Association between circulating irisin levels and the promotion of insulin resistance during
the weight maintenance period after a dietary weight-lowering program in obese patients. Metabolism 2014,
63, 520–531. [CrossRef] [PubMed]
73. Huerta, A.E.; Prieto-Hontoria, P.L.; Fernandez-Galilea, M.; Sainz, N.; Cuervo, M.; Martinez, J.A.;
Moreno-Aliaga, M.J. Circulating irisin and glucose metabolism in overweight/obese women: Effects
of α-lipoic acid and eicosapentaenoic acid. J. Physiol. Biochem. 2015, 71, 547–558. [CrossRef] [PubMed]
74. Weisberg, S.P.; McCann, D.; Desai, M.; Rosenbaum, M.; Leibel, R.L.; Ferrante, A.W. Obesity is associated with
macrophage accumulation in adipose tissue. J. Clin. Investig. 2003, 112, 1796–1808. [CrossRef] [PubMed]
75. Jiao, P.; Chen, Q.; Shah, S.; Du, J.; Tao, B.; Tzameli, I.; Yan, W.; Xu, H. Obesity-related upregulation of
monocyte chemotactic factors in adipocytes: Involvement of nuclear factor-kappaB and c-Jun NH2-terminal
kinase pathways. Diabetes 2009, 58, 104–115. [CrossRef] [PubMed]
76. Li, C.; Xu, M.M.; Wang, K.; Adler, A.J.; Vella, A.T.; Zhou, B. Macrophage polarization and meta-inflammation.
Transl. Res. 2018, 191, 29–44. [CrossRef]
77. Rogero, M.; Calder, P. Obesity, inflammation, toll-like receptor 4 and fatty acids. Nutrients 2018, 10, 432.
[CrossRef]
78. Lumeng, C.N.; Bodzin, J.L.; Saltiel, A.R. Obesity induces a phenotypic switch in adipose tissue macrophage
polarization. J. Clin. Investig. 2007, 117, 175–184. [CrossRef]
79. Mazur-Bialy, A.I.; Bilski, J.; Pochec, E.; Brzozowski, T. New insight into the direct anti-inflammatory activity
of a myokine irisin against proinflammatory activation of adipocytes: Implication for exercise in obesity.
J. Physiol. Pharmacol. 2017, 68, 243–251.
80. Choi, C.H.J.; Cohen, P. Adipose crosstalk with other cell types in health and disease. Exp. Cell Res. 2017, 360,
6–11. [CrossRef]
81. Maffei, M.; Halaas, J.; Ravussin, E.; Pratley, R.E.; Lee, G.H.; Zhang, Y.; Fei, H.; Kim, S.; Lallone, R.;
Ranganathan, S.; et al. Leptin levels in human and rodent: Measurement of plasma leptin and ob RNA in
obese and weight-reduced subjects. Nat. Med. 1995, 1, 1155–1161. [CrossRef]
82. do Carmo Martins, M.; Faleiro, L.L.; Fonseca, A. Relationship between leptin and body mass and metabolic
syndrome in an adult population. Rev. Port. Cardiol. 2012, 31, 711–719.
83. Gutierrez-Repiso, C.; Garcia-Serrano, S.; Rodriguez-Pacheco, F.; Garcia-Escobar, E.; Haro-Mora, J.J.;
Garcia-Arnes, J.; Valdes, S.; Gonzalo, M.; Soriguer, F.; Moreno-Ruiz, F.J.; et al. FNDC5 could be regulated by
leptin in adipose tissue. Eur. J. Clin. Investig. 2014, 44, 918–925. [CrossRef]
84. Hou, N.; Liu, Y.; Han, F.; Wang, D.; Hou, X.; Hou, S.; Sun, X. Irisin improves perivascular adipose tissue
dysfunction via regulation of the heme oxygenase-1/adiponectin axis in diet-induced obese mice. J. Mol. Cell.
Cardiol. 2016, 99, 188–196. [CrossRef] [PubMed]
85. Han, F.; Zhang, S.; Hou, N.; Wang, D.; Sun, X. Irisin improves endothelial function in obese mice through the
AMPK-eNOS pathway. Am. J. Physiol. Heart Circ. Physiol. 2015, 309, H1501–H1508. [CrossRef] [PubMed]
86. Provatopoulou, X.; Georgiou, G.P.; Kalogera, E.; Kalles, V.; Matiatou, M.A.; Papapanagiotou, I.; Sagkriotis, A.;
Zografos, G.; Gounaris, A. Serum irisin levels are lower in patients with breast cancer: Association with
disease diagnosis and tumor characteristics. BMC Cancer 2015, 15, 898. [CrossRef] [PubMed]
87. Zhang, Z.P.; Zhang, X.F.; Li, H.; Liu, T.J.; Zhao, Q.P.; Huang, L.H.; Cao, Z.J.; He, L.M.; Hao, D.J. Serum irisin
associates with breast cancer to spinal metastasis. Medicine 2018, 97, 17. [CrossRef] [PubMed]
88. Altay, D.U.; Keha, E.E.; Karagüzel, E.; Menteşe, A.; Yaman, S.O.; Alver, A. The Diagnostic Value of
FNDC5/Irisin in Renal Cell Cancer. Int. Braz. J. Urol. 2018, 44, 734–739. [CrossRef] [PubMed]
89. Gannon, N.P.; Vaughan, R.A.; Garcia-Smith, R.; Bisoffi, M.; Trujillo, K.A. Effects of the exercise-inducible
myokine irisin on malignant and non-malignant breast epithelial cell behavior in vitro. Int. J. Cancer 2015,
136, E197–E202. [CrossRef] [PubMed]
90. Liu, J.; Song, N.; Huang, Y.; Chen, Y. Irisin inhibits pancreatic cancer cell growth via the AMPK-mTOR
pathway. Sci. Rep. 2018, 8, 15247. [CrossRef] [PubMed]
91. Kong, G.; Jiang, Y.; Sun, X.; Cao, Z.; Zhang, G.; Zhao, Z.; Zhao, Y.; Yu, Q.; Cheng, G. Irisin reverses the
IL-6 induced epithelial-mesenchymal transition in osteosarcoma cell migration and invasion through the
STAT3/Snail signaling pathway. Oncol. Rep. 2017, 38, 2647–2656. [CrossRef] [PubMed]
Medicina 2019, 55, 485 14 of 14

92. Shao, L.; Li, H.; Chen, J.; Song, H.; Zhang, Y.; Wu, F.; Wang, W.; Zhang, W.; Wang, F.; Li, H.;
et al. Irisin suppresses the migration, proliferation, and invasion of lung cancer cells via inhibition
of epithelial-to-mesenchymal transition. Biochem. Biophys. Res. Commun. 2017, 485, 598–605. [CrossRef]
[PubMed]
93. Moon, H.S.; Mantzoros, C.S. Regulation of cell proliferation and malignant potential by irisin in endometrial,
colon, thyroid and esophageal cancer cell lines. Metabolism 2014, 63, 188–193. [CrossRef] [PubMed]
94. Shi, G.; Tang, N.; Qiu, J.; Zhang, D.; Huang, F.; Cheng, Y.; Ding, K.; Li, W.; Zhang, P.; Tan, X. Irisin stimulates
cell proliferation and invasion by targeting the PI3K/AKT pathway in human hepatocellular carcinoma.
Biochem. Biophys. Res. Commun. 2017, 493, 585–591. [CrossRef] [PubMed]
95. Hoja-Łukowicz, D.; Przybyło, M.; Duda, M.; Pocheć, E.; Bubka, M. On the trail of the glycan codes stored in
cancer-related cell adhesion proteins. Biochim. Biophys. Acta Gen. Subj. 2017, 1861, 3237–3257. [CrossRef]
[PubMed]
96. Pinho, S.S.; Reis, C.A. Glycosylation in cancer: Mechanisms and clinical implications. Nat. Rev. Cancer 2015,
15, 540–555. [CrossRef] [PubMed]
97. Alter, G.; Ottenhoff, T.H.M.; Joosten, S.A. Antibody glycosylation in inflammation, disease and vaccination.
Semin. Immunol. 2018, 39, 102–110. [CrossRef]

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