European Journal of Pharmacology 785 (2016) 207–214

Contents lists available at ScienceDirect

European Journal of Pharmacology

journal homepage: www.elsevier.com/locate/ejphar

Fatty acids and chronic low grade inflammation associated with

obesity and the metabolic syndrome
Aoife A. Cooke, Ruth M. Connaughton, Claire L. Lyons, Aoibheann M. McMorrow,
Helen M. Roche n
Nutrigenomics Group, UCD Conway Institute for Biomolecular and Biomedical Research & UCD Institute of Food & Health, School of Public Health, Phy-
siotherapy and Sports Science, University College Dublin, Belfield, Dublin 4, Ireland

art ic l e i nf o a b s t r a c t

Article history: The metabolic syndrome is a group of obesity associated metabolic conditions that result in increased
Received 3 December 2015 risk of cardiovascular disease and type 2 diabetes. Global increases in obesity rates have led to an in-
Received in revised form crease in metabolic syndrome resulting in a demand for increased understanding of the mechanisms
2 April 2016
involved. This review examines the relationship between adipose tissue biology, lipid metabolism and
Accepted 11 April 2016
Available online 12 April 2016
chronic low grade inflammation relating to obesity and insulin resistance.
& 2016 Published by Elsevier B.V.
Keywords:
Metabolic syndrome
Obesity
Fatty acids
Inflammation

1. Introduction often associated with metabolic abnormalities including type

First described in the 1920s as a combination of metabolic
disturbances, the scientific community has somewhat struggled to
define the Metabolic Syndrome (MetS) (Kylin, 1923). In the 1940s
it was recognised that upper body adiposity was the type most
Abbreviations: AMPK, 5′ Adenosine Monophosphate-Activated Protein Kinase;
ATGL, Adipose Triacylglycerol Lipase; BAT, Brown Adipose Tissue; BMI, Body Mass
Index; CVD, Cardiovascular Disease; DAG, Diacylglycerol; DHA, Docosahexaenoic
Acid; DIO, Diet Induced Obesity; EPA, Eicosapentaenoic Acid; ERK, Extra-Cellular
Signal Related Kinase; GLUT-4, Glucose Transporter Type
4; HDL, High-Density
Lipoprotein; HFD, High Fat Diet; HSL, Hormone Sensitive Lipase; Ig, Im-
munoglobulin; IKK-β, Inhibitor of Nuclear Factor Kappa-B Kinase Subunit –Beta; IL,
Interleukin; IL-1R-1, Interleukin-1 Receptor-1; IRS, Insulin Receptor Substrate;
IκBα, Nuclear Factor Kappa Light Polypeptide Chain Gene Enhancer in B-Cells In-
hibitor, Alpha; JAK, Janus Kinase; JNK, c-Jun-N-Terminal Protein Kinase; LPL, Lipo-
protein Lipase; MAPK, Mitogen Activated Protein Kinase; MCP-1, Monocyte Che-
moattractant Protein
1; MetS, Metabolic Syndrome; MUFA, Monounsaturated
Fatty Acid; (n
3 LCPUFA), n
3 Long Chain Polyunsaturated Fatty Acid; NAFLD,
Non-Alcoholic Fatty Liver Disease; NFκB, Nuclear Factor Kappa Light Chain En-
hancer of β-Cells; PKC, Protein Kinase C; PKC-θ, Protein Kinase C-θ; PKR, Protein
Kinase R; PPAR-γ, Peroxisome Proliferator Activated Receptor γ; PUFA, Poly-
unsaturated Fatty Acid; SAT, Subcutaneous Adipose Tissue; SFA, Saturated Fatty
Acid; SOCS-3, Suppressor of Cytokine Signalling-3; STAT-3, Signal Transducer and
Activator of Transcription-3; SVF, Stromal Vascular Fraction; T2D, Type 2 Diabetes;
TAG, Triacylglycerol; Th-1, T Helper
1; TLR-4, Toll like Receptor-4; TNFR-1, Tu-
mour Necrosis Factor Receptor – 1; TNF-α, Tumour Necrosis Factor – α; Treg, T-
Regulatory; VAT, Vascular Adipose Tissue; WAT, White Adipose Tissue
n
Corresponding author.
E-mail address: helen.roche@ucd.ie (H.M. Roche).
2 diabetes (T2D) and cardiovascular disease (CVD) (Vague, 1947).
In 1988 Gerald Reaven used the term Syndrome X to describe a
collection of symptoms including resistance to insulin-stimulated
glucose uptake, glucose intolerance, hyperinsulinemia, increased
very-low-density lipoprotein triglyceride or triacylglycerol (TAG),
decreased high-density lipoprotein (HDL) cholesterol and hy-
pertension. However, obesity was notably absent from this list
(Reaven, 1988). The most recent formal definition for MetS avail-
able identifies raised blood pressure, elevated TAG, lowered HDL
cholesterol, raised fasting glucose, and central obesity as its cri-
teria (Alberti et al., 2009). Individuals with MetS experience at
least a fivefold increased risk of developing T2D and a twofold
increased risk of CVD, with profound implications for morbidity
and quality of life (Eckel et al., 2010, 2005).
The global increase in obesity rates is of grave concern. As of
2011, the number of individuals classified as overweight is greater
than those classified as undernourished, with more than 1.7 billion
overweight individuals worldwide (Chawla et al., 2011). Increasing
obesity rates are closely linked to greater prevalence of the MetS,
non-alcoholic fatty liver disease (NAFLD), T2D and CVD (Hong
et al., 2015; Sung et al., 2012; Targher and Arcaro, 2007; Targher,
2007; Targher et al., 2006). These statistics represent an enormous
stress on global health services. It is imperative then that the
mechanisms behind obesity and MetS are understood, in order to
improve both preventative measures and outcomes. This review
will focus on the inter-relationship between fatty acids and
chronic low-grade inflammation in adipose tissue, which in turn

http://dx.doi.org/10.1016/j.ejphar.2016.04.021
0014-2999/& 2016 Published by Elsevier B.V.
208 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214

Fig. 1. The inter-relationship between fatty acids, inflammation and insulin signalling - Cellular cross talk between macrophages and adipocytes. Fatty acids and their
derivatives have been shown to modulate inflammatory pathways in immune cells. This results in increased pro-inflammatory cytokine secretion from immune cells such as
macrophages and T cells which in turn act on adipocytes to disrupt insulin signalling. (This figure was prepared using the Servier medical art website http:// www.servier.fr/
servier-medical-art.).

modulates insulin signalling (Fig. 1). Given the importance of lipid
metabolism in this paradigm, our perspective will focus on whe-
ther different fatty acids may mediate differential effects on adi-
pose tissue inflammation and insulin resistance, within the
broader context of the MetS and T2D risk.
2. Adipose tissue
Obesity is characterised by excess adipose tissue, which itself
can be categorised into two types: brown adipose tissue (BAT) and
white adipose tissue (WAT) (Rosenwald and Wolfrum, 2014). BAT
is known to play a role in thermogenic processes (Nedergaard
et al., 2007). Until recently, BAT was thought to be present only in
new-born humans. However, recent work has suggested a role for
BAT in thermoregulatory processes in adults (Nedergaard et al.,
2007). In contrast, WAT plays a significant role in regulating en-
ergy metabolism and storing excess energy as TAG (Schweiger and
Schreiber, 2006).
Adipocytes represent the primary cellular component of WAT.
The lipid stored within adipocytes is in constant flux and readily
adapts to fluctuating nutritional demand (Lafontan and Langin,
2009). In the postprandial state, lipoprotein lipase (LPL), located
within the endothelial lining of blood vessels hydrolyses circulat-
ing TAG. This process releases free fatty acids which are taken up
by adipocytes and re-esterified to generate TAG for storage. In
contrast, in times of energy deficit or increased energy require-
ments, TAG undergoes lipolysis, wherein it is hydrolysed by adi-
pose triacylglycerol lipase (ATGL) and hormone sensitive lipase
(HSL) to release free fatty acids for β-oxidation (Lafontan and
Langin, 2009).
During persistent positive energy balance, WAT is forced to
expand to facilitate increased TAG storage (Virtue and Vidal-Puig,
2010). Depending on its location, increased adipose tissue expan-
sion can result in considerable adipose tissue dysfunction, which is
characterised by decreased insulin sensitivity (Hotamisligil et al.,
1993), increased immune cell infiltration (Hotamisligil, 2006), fi-
brosis (Sun et al., 2013), hypoxia (Sun et al., 2013) and increased
intracellular and systemic free fatty acid flux (Yu and Ginsberg,
2005). Expansion of WAT is peroxisome proliferator- activated
receptor-γ (PPAR-γ) dependent and occurs via an increase in adi-
pocyte cell size (hypertrophy), cell number (hyperplasia) or both
(Jo et al., 2009). It is now known that increased hyperplasia is
linked to a more favourable phenotype, which likely reflects less
adipose tissue inflammation (Lundgren et al., 2007).
In addition to cell morphology, distribution of excess adipose
tissue has a significant impact on pathology, wherein increased
visceral adipose tissue (VAT) to a greater extent than subcutaneous
adipose tissue (SAT) is associated with MetS development
(Després and Lemieux, 2006). Additionally, SAT is rich in pre-
adipocytes prone to rapid replication, which increases its buffering
capacity (Tchkonia et al., 2005). Vidal-Puig and colleagues hy-
pothesised that individuals reach a limit of SAT expansion, after
which excess TAG can no longer be facilitated within the sub-
cutaneous region (Virtue and Vidal-Puig, 2008). This inability of
SAT to buffer macronutrient surplus promotes lipotoxicity,
wherein excess energy is directed towards the VAT depot as well
as to non-adipose locations, such as skeletal muscle, the liver and
 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214
the pancreas (Virtue and Vidal-Puig, 2008). This concept is known
as overspill and is detrimental to organ signalling and function-
ality owing to the fact that increased lipid uptake is not matched
by increased β-oxidation (Tumova et al., 2015).
Furthermore, impaired insulin signalling reduces postprandial
inhibition of lipolysis in response to insulin (Basu et al., 2001).
Thus, insulin resistance is associated with increased and persistent
free fatty acid flux from the adipose tissue into the circulation.
Visceral adipocytes in particular have been shown to be the pri-
mary contributors of free fatty acids to circulation, attributable to
higher basal lipolytic activity in VAT relative to SAT (Arner, 1995).
Furthermore, obese subjects have demonstrated reduced ability to
clear free fatty acids following a high-fat meal (Lewis and O’Meara,
1990). Thus, adipose tissue dysfunction gives rise to increased
systemic free fatty acids owing to limited SAT buffering capacity,
increased endogenous free fatty acid release and decreased ability
to clear exogenous free fatty acids.
3. Adipose tissue immune cell infiltration
While adipocytes are the largest component of adipose tissue,
the stromal vascular fraction (SVF), although smaller, plays a key
role in adipose function. The SVF of the adipose is where immune
cells, such as macrophages (10–15%), CD3 þ T cells (14%; 94% of
which are CD4 þ or CD8 þ ), mast cells, eosinophils (4–5%) and B
cells reside (Nishimura et al., 2009; Weisberg et al., 2003; Winer
et al., 2011; Wu et al., 2011). In the lean insulin sensitive state,
anti-inflammatory T regulatory (Treg) and M2 macrophages are
the prominent cell types. Resident immune cells have house-
keeping roles, including, but not limited to, clearance of apoptotic
adipocytes, extracellular remodeling, angiogenesis and adipogen-
esis (Schipper et al., 2012).
As the VAT expands, immune cells infiltrate the tissue to form
crown-like structures around necrotizing adipocytes, in order to
clear cellular debris and perform an adaptive function (Nishimura
et al., 2009; Xu et al., 2003). In addition the immune cell compo-
sition and phenotype is altered in the obese setting contributing to
adipose tissue dysfunction and insulin resistance. Obesity is as-
sociated with a state of chronic low-grade inflammation and this
provides signals that drive a pro-inflammatory and persistent
immune response (which is discussed in the next section). In ob-
ese adipose, immune cells are under increased demand associated
with adipose expansion. Adipose tissue macrophages and mast cell
localization around microvessels provide angiogenic support for
adipose tissue development and leukocyte infiltration into the
expanding depot. An increase in CD31 þ endothelial cells is also
observed in dietary induced obesity (DIO) to facilitate increased
angiogenesis (Nishimura et al., 2009). Extracellular remodeling is
undertaken by the mast cells to facilitate growth (Liu et al., 2009).
The influx of immune cells begins with CD8 þ T cells as early as
4 weeks on a high-fat diet (HFD), followed by macrophage re-
cruitment, with a decrease in CD4 þhelper and Treg cells occurring
at later time points (Nishimura et al., 2009). Treatment with a CD8
specific antibody, in conjunction with a HFD, results in reduced M1
polarization with no change to M2 or CD4 þ T cell numbers; de-
monstrating the interplay between the different immune cell
types. Amelioration of the inflammatory response and insulin re-
sistant state is observed following inhibition of CD8 þ T cell action,
even in a pre-established inflammatory state, indicating T cell in-
volvement in both the propagation and maintenance of adipose
tissue inflammation (Nishimura et al., 2009). When reduction of
Treg cells is induced, insulin levels increase (Feuerer et al., 2009).
Treg cells produce large amounts of the anti-inflammatory cyto-
kine interleukin (IL)
10, which itself has insulin sensitizing effects
(Lumeng et al., 2008).
209
T cells are necessary for the activation, differentiation and mi-
gration of macrophages (Nishimura et al., 2009). Adipose tissue
macrophage content increases from 10% to 15% in lean adipose to
45–60% within obese adipose (Weisberg et al., 2003). In a micro-
array analysis of adipose tissue, a third of the transcripts that were
positively correlated with body mass belonged to macrophages.
Adipocyte area in multiple adipose depots is a strong positive
predictor of F4/80 þ macrophage number, a result also observed
with increasing body mass index (BMI) in human SAT (Weisberg
et al., 2003). For example, CD45.2 mice transplanted with CD45.1
bone marrow, displayed 80% of F4/80 þ macrophages from the
CD45.1 donor within adipose tissue following 6 weeks of HFD,
demonstrating the influx of immune cells from the periphery
(Weisberg et al., 2003). Chemokines, such as monocyte chemoat-
tractant protein 1 (MCP-1) play a role in the recruitment of mac-
rophages to the adipose tissue in both HFD and genetic models of
obesity (Kamei et al., 2006). Once the immune cells populate the
adipose tissue, their secreted cytokines attract more immune cells
into the local area. Resident macrophages undergo a phenotypic
switch from the anti-inflammatory M2 to the pro-inflammatory
M1 adipose tissue macrophages, through cytokine and immune
cell action (Lumeng et al., 2008). Eosinophils maintain macro-
phages in an M2 state in an IL-4/IL-13 dependent manner but their
numbers decrease with increasing adiposity (Wu et al., 2011),
therefore an M1 phenotype is the dominant immunophenotype in
obesity. Eosinophil-deficient mice display obesity with glucose
intolerance, indicating their role in metabolic homeostasis. The
innate immune mast cell has been implicated in DIO as their
numbers increase with obesity, inflammation and insulin re-
sistance (Winer et al., 2011). B cells increase inflammatory med-
iators, such as secretion of tumor necrosis factor-α (TNF-α); in-
fluence CD8 þ and T helper-1 (Th1) T cell activation; and produce
immunoglobulin (Ig) antibodies, which are involved in macro-
phage pro-inflammatory polarization. B cell knockout models are
protected against disrupted glucose homeostasis despite equiva-
lent levels of obesity compared to their control counterparts
(Winer et al., 2011). This demonstrates that both the innate and
adaptive immune systems play a pivotal role in the development
of insulin resistance and MetS, mainly through the release of pro-
inflammatory cytokines.
4. Localised inflammatory mediators
Free fatty acids arising from inflamed adipose tissue activate a
host of intracellular signalling cascades that sense and commu-
nicate adipocyte nutrient overload. This occurs through the acti-
vation of the c-Jun N-terminal protein kinases (JNK), inhibitor of
nuclear factor kappa-B kinase subunit beta (IKK-β), and protein
kinase R (PKR) signalling pathways, all of which are involved in the
inflammatory response (Hirosumi et al., 2002; Nakamura et al.,
2010; Yuan et al., 2001). Activation of these signalling pathways
results in inhibition of the insulin signalling pathway by serine
phosphorylation of insulin receptor substrate (IRS) proteins
(Aguirre et al., 2002; Hotamisligil et al., 1996). Furthermore, JNK
and IKK-β activation initiates a paracrine inflammatory response
involving cytokines, such as TNF-α and IL-1β. These can broadcast
intracellular conditions to other adipocytes and importantly to
resident leukocytes in the immediate microenvironment (Ode-
gaard and Chawla, 2013).
TNF-α, the first pro-inflammatory cytokine linked to obesity-
associated insulin resistance activates a number of serine kinases,
such as JNK and IKKβ, leading to serine phosphorylation of IRS-1
(Gao et al., 2002; Hirosumi et al., 2002; Hotamisligil et al., 1993).
This is in contrast to tyrosine phosphorylation of IRS-1 in an in-
sulin sensitive state. Moreover, TNF-α, in conjunction with IL-6,
210 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214
increases the secretion of suppressor of cytokine signalling-3
(SOCS-3) proteins, which bind to insulin receptors and attenuate
phosphorylation of IRS proteins thus impairing insulin signalling
(Emanuelli et al., 2000; Ueki et al., 2004). Demonstrating the im-
portant role TNF-α plays in obesity-induced insulin resistance,
obese rodents which had been administered a TNF-α neutralising
antibody exhibited reduced hyperinsulinemia (Hotamisligil et al.,
1993). Additionally, whole body deletion of TNF-α or its corre-
sponding receptor tumour necrosis factor receptor-1 (TNFR1)
partially protects mice from obesity induced insulin resistance
(Uysal et al., 1997).
With respect to IL-6, there is conflicting data regarding its role
in insulin resistance (Pedersen and Febbraio, 2007). IL-6 is se-
creted by adipose tissue, as well as skeletal muscle and liver, sig-
nalling via the janus kinase (JAK)/signal transducer and activator
of transcription (STAT) and mitogen activated protein kinase
(MAPK) pathways. In adipose tissue, IL-6 reduces glucose uptake
in adipocytes, by reducing insulin-stimulated tyrosine phosphor-
ylation of IRS-1 while concurrently promoting serine phosphor-
ylation of IRS-1 (Fasshauer et al., 2003; Rotter et al., 2003). How-
ever in skeletal muscle the role played by IL-6 can vary. IL-6 en-
hances fat oxidation in skeletal muscle via activation of 5′ ade-
nosine monophosphate activated protein kinase (AMPK) (Carey
et al., 2006). Furthermore, Wolsk et al. have demonstrated that IL-
6 enhances fat metabolism, specifically lipolysis, in skeletal muscle
but not in adipose (Wolsk et al., 2010). In contrast, other studies
have associated skeletal muscle wastage with chronic systemic IL-
6 levels (Fearon et al., 2012). Thus any positive effects of IL-6 in
skeletal muscle are associated with transient production, rather
than chronic exposure (Munoz-Canoves et al., 2013).
IL-1 are a pro-inflammatory family of cytokines that play a key
role in insulin resistance (Osborn and Olefsky, 2012). Consisting of
two major proteins, IL-1α and IL-1β, both display similar biological
properties but minimal sequence homology (Dinarello, 2009).
Recently, studies have established that IL-1β requires the activa-
tion of the NLRP3 inflammasome, a protein complex, to produce
its biologically active mature form (Tschopp and Schroder, 2010).
Activation of the NLRP3 inflammasome leads to the cleavage of
pro-caspase to caspase-1, which in turn cleaves the inactive pro-IL-
1β to the mature, biologically potent IL-1β (Koenen et al., 2011).
This is distinct to IL-1α, which is produced in its biologically active
form (Dinarello, 2009). IL-1β is considerably the more potent in-
flammatory cytokine and is involved in many inflammatory dis-
eases by initiating and potentiating immune and inflammatory
responses (Dinarello, 2009). In 3T3L1 adipocytes, IL-1β decreases
glucose transport via inhibition of IRS-1 expression, by activating
the inflammatory kinases MAPK and extra-cellular signal related
kinase (ERK) (Jager et al., 2007). Furthermore, treatment of adi-
pocytes with IL-1β reduced insulin-stimulated glucose uptake by
decreasing expression and translocation of glucose transporter
type-4 (GLUT-4) to the plasma membrane (He et al., 2006). HFD
feeding induces NLRP3 inflammasome activation, via cleavage and
activation of caspase-1, resulting in secretion of IL-1β (Martinon
et al., 2002; Reynolds et al., 2012). Correspondingly, mice lacking
the IL-1 receptor 1(IL-1R1) are protected from HFD-induced glu-
cose intolerance (De Roos et al., 2009; McGillicuddy et al., 2011).
Conversely anti-inflammatory cytokines, such as IL-10, are also
secreted by immune cells and may have insulin sensitising po-
tential. IL-10, produced by monocytes, M2 adipose tissue macro-
phages, dendritic cells, T cells and B cells, signals via the IL-10
receptor activating the JAK/STAT pathway (Saraiva and O’Garra,
2010). Protein levels of NFκB inhibitor-α (IκBα), an inhibitor of
nuclear factor kappa light chain enhancer of B cells (NFκB), are
preserved following treatment with IL-10, attenuating NFκB
binding as well as TNF-α levels (Shames et al., 1998). Furthermore
IL-10 is a potent inhibitor of pro-inflammatory cytokines and
chemokines such as TNF-α and IL-6; and in turn their detrimental
biological effect on target cells, such as adipocytes (Hong et al.,
2009). Other intracellular events affected by IL-10 involve nuclear
translocation of signal transducer and activator of transcription 3
(STAT3) and suppressor of cytokine signalling 3 (SOCS3). Induction
of SOCS3 by IL-10 inhibits signalling that is initiated by IL-6 via the
IL-6 signal transducer (gp130) (Niemand et al., 2003). In adipocyte
cell culture models, pre-treatment with IL-10 attenuated the in-
sulin desensitizing effect of macrophage conditioned media, thus
by modulating or attenuating inflammation, IL-10 may potentiate
insulin sensitivity (Oliver et al., 2012).
5. Dysregulated fatty acid metabolism and metabolic
homeostasis
The relationship between obesity induced inflammation and
lipid dysregulation is bidirectional. It is important to acknowledge
that several components of dysregulated lipid metabolism and
chronic low-grade inflammation play a causal role in the patho-
genesis and progression of insulin resistance. However, several
abnormalities of lipid metabolism associated with insulin re-
sistance may simply reflect the insulin resistant state. On the
causality side, Schulman's team have demonstrated very elegantly
the concept of lipotoxicity, wherein overspill of free fatty acids
leading to ectopic lipid accumulation in skeletal muscle and the
liver, can lead to insulin resistance (Kim et al., 2000; Nagle et al.,
2007; Samuel and Shulman, 2012). As reviewed extensively, there
are two principle routes wherein fatty acids/lipids directly impede
insulin signalling (Johnson and Olefsky, 2013). Firstly, circulating
free fatty acids can activate cell-signalling pathways, to impede
insulin action (Glass and Olefsky, 2012). Secondly, imbalanced lipid
metabolism and/or metabolic derivatives thereof, lead to accu-
mulation of intracellular lipid products, including diacylglycerol
(DAG), in both hepatic and skeletal tissues wherein lipotoxicity
causes insulin resistance (Jornayvaz and Shulman, 2012). DAG in
turn activates protein kinase Cε (PKC), which then impedes tyr-
osine phosphorylation of IRS
1 and
2 to precipitate the hepatic
insulin resistance that underlies NAFLD associated with obesity,
MetS and T2D (Jornayvaz and Shulman, 2012). Taken together,
these routes demonstrate that lipotoxicity plays a causal role in
the MetS.
Ceramides are another critical lipid intermediate derived from
saturated fatty acid (SFA), which impede AKT, the activity of which
is crucial to insulin signalling (Holland et al., 2011; Powell et al.,
2004). There is also strong evidence that an excessive fatty acid
insult upon the mitochondria overwhelm their capacity for β-
oxidation, which generates partially oxidised acylcarnitines that
also have adverse effects on insulin signalling (Muoio and Neufer,
2012). Hepatocellular DAG and acylcarnitine content probably re-
flects the balance between lipogenesis and mitochondrial fatty
acid oxidation, the combination of which affects the degree of
hepatic insulin resistance. Furthermore, Holland and colleagues
showed the interconnections between ceramide biology and in-
flammation. Activation of toll like receptor-4 (TLR4)-dependent
inflammatory pathways by SFA augments ceramide biosynthesis
(Holland et al., 2011). This interplay between fatty acids and in-
flammatory processes occurs at a number of other metabolic-in-
flammatory hubs, as discussed in the next section.
6. Different fatty acids ¼different metabolic effects?
A core theme of our recent research has addressed whether
different dietary fatty acids may have a variable impact on meta-
bolic homeostasis. We have some interesting insights; however
 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214
the full nature of the inter-relationship between fatty acid, low
grade chronic inflammation and obesity related insulin resistance
is not fully understood. Fatty acids are ubiquitous in the diet,
distributed throughout the body and play a key role in lipid me-
tabolism. Fatty acids may be present in their free form, as free fatty
acids. However, they are more abundantly represented in several
complex lipid forms, such as TAG, phospholipids or as cholesterol
esters. Exogenously these fatty acid-enriched complex lipid spe-
cies are present in lipid containing food. Dietary fat is principally
composed of TAG, with lesser amounts of phospholipids and
cholesterol esters. Endogenous lipid metabolism is principally
determined by the liver, with interactions between adipose tissue
and skeletal muscle, the former a storage site representing an
important endogenous source of fatty acids and the latter being a
catabolic site. Lipids play a fundamental role in intra-organ and
cellular biology as lipid storage molecules (TAG) and cellular
membrane components (phospholipids). Fatty acids can also act as
potent intracellular second messenger signalling molecules
wherein they may be present in DAG ceramides and other lipid
derivatives.
There is a large body of evidence that shows that obesity per se,
induced by HFD, instigates an inflammatory cascade that impedes
insulin signalling, as previously reviewed (Murphy et al., 2014). For
example, Reynolds et al. demonstrated that this priming and ac-
tivation of inflammation via TLR4 and NLRP3 by dietary SFA is
evident in bone marrow dendritic cells, in a manner that is con-
current with that of adipose tissue inflammation activation (Rey-
nolds et al., 2012). In the case of the liver, this paradigm is not
necessarily correct. Galbo et al. demonstrated that TLR-4 mediated
hepatic ceramide synthesis signalling is not directly required for
SFA or polyunsaturated fatty acid (PUFA) induced hepatic insulin
resistance. It is of note that this study investigated the short-term
(3 days) impact of dietary SFA versus PUFA, wherein both SFA and
PUFA-HFD led to hepatic DAG accumulation, PKCε activation and
impaired insulin signaling (Galbo et al., 2013).
Our recent work has focused on SFA mediated priming and
activation of the NLRP3 inflammasome resulting in IL-1β-medi-
ated adipose tissue inflammation and insulin resistance. Interest-
ingly replacement of SFA for monounsaturated fatty acid (MUFA)
in HFD reduced adipose tissue pro-IL-1β priming and attenuated
insulin resistance in an AMPK mediated fashion (Finucane et al.,
2015). Despite being obese, MUFA-HFD-fed mice displayed im-
proved insulin sensitivity coincident with reduced pro-IL-1β
priming, attenuated adipose IL-1β secretion and sustained adipose
activation of the nutritional sensor AMPK. Adipose tissue mor-
phology was markedly different between diets, which we suspect
had a very significant physiological impact upon the whole body
insulin and glucose homeostasis. We speculate that greater acti-
vation of IL-1β in the adipose milieu by SFA affected the adipo-
genic potential of preadipocytes creating a hypertrophic adipose
morphology. In contrast despite equal adipose tissue weight, the
MUFA-HFD mice displayed hyperplasic adipose tissue with pre-
served insulin sensitivity, which we attributed to attenuated
NLRP3 mediated IL-1β activation in adipose. Whilst this work is
very interesting in terms of the relative impact of SFA versus
MUFA, we are as yet unsure the extent to which this paradigm
relates to man. Previous work from our group has demonstrated
that increasing SFA but not MUFA induced insulin resistance in
T2D subjects suggesting that a HFD rich in MUFA are not as de-
leterious. Furthermore we examined data from subjects with
MetS, which demonstrated that when subjects with a habitual
high MUFA intake underwent a MUFA dietary intervention they
had improved insulin response to glucose and improved first-
phase insulin response. This was in contrast to high-SFA habitual
consumers who experienced no such improvement (Finucane
et al., 2015). The prospective evidence is strong with respect to the
211
adverse effects of high SFA diets as reviewed in detail previously
(Phillips et al., 2008). However we do not yet fully understand if/
how different fatty acids mediate their adverse effect. Also it is
unlikely that this effect can be ascribed only to IL-1β mediated
inflammation and insulin resistance. For example, it has been
shown that overfeeding SFA and PUFA has different effects on
hepatic and VAT lipid accumulation (Rosqvist et al., 2014). Also
different dietary lipids may affect skeletal muscle fatty acid
handling (Jans et al., 2012b).
Other recent and exciting developments in this field include
the discovery of novel endogenous lipid species which have anti-
diabetic and anti-inflammatory effects. Yore et al. demonstrated
enhanced adipose lipogenesis, greater glucose tolerance and anti-
inflammatory properties, in spite of obesity. This was attributable
to elevated concentrations of a novel class of lipids in GLUT-4
transgenic animals. These novel lipids were characterised by
branched ester linkage between a fatty acid and a hydroxy-fatty
acid and are referred to as FAHFAs. The most dramatically upre-
gulated FAHFA in GLUT-4 transgenic mice was palmitic-acid-hy-
droxyl-stearic acid (PAHSA). PAHSAs are synthesised in vivo and
regulated by both fasting and HFD feeding. Importantly, PAHSA
levels were lower in serum and adipose tissue of insulin resistant
subjects. Yore et al. demonstrated that PAHSAs had multiple ef-
fects that resulted in improved glucose-insulin homeostasis, which
suggests that restoration of PAHSA levels in patients with insulin
resistance may have potential beneficial effects on metabolism. In
animal studies, oral administration of PAHSA to insulin-resistant
mice on a HFD improved glucose tolerance (Yore et al., 2014). In-
terestingly PAHSA signalled via G protein-coupled receptor 120
(GPR120). GPR120 was also identified as an important signalling
sensor/receptor that mediated the anti-inflammatory, insulin-
sensitising effects of the two n-3 long chain PUFA ((n
3 LCPUFA))
eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Oh
et al., 2010). However, in this context the dietary doses of (n
3
LCPUFA) were supra-physiological and not necessarily translatable
to humans. Nevertheless, GPR120 is an important control point in
the integration of anti-inflammatory insulin sensitisation and the
focus of on-going research as a therapeutic target within the
context of obesity related inflammation and insulin resistance in
T2D (Oh et al., 2014). There are several other fatty acid sensitive,
molecular targets at the inter-section of metabolic-inflammation,
including ChREBP, NF-κB, PPARγ which for the sake of brevity are
not discussed here, but are well documented (Benhamed et al.,
2012; Herman et al., 2012; Murphy et al., 2014; Osborn and
Olefsky, 2012). Whilst we have focused on fatty acids; it is im-
portant to note that there are additional non-lipid nutrients with
anti-inflammatory potential, such as lycopene, ECGC, vitamin C
and resveratrol, as reviewed in detail elsewhere (Connaughton
et al., 2016; Schrauwen and Timmers, 2014).
7. Translational perspectives – efficacy of dietary fatty acid
modulation in humans?
In the acute setting, SFA are more potent than MUFA or PUFA in
inducing an insulin resistant or glucose intolerant state (Jans et al.,
2012a; Xiao et al., 2006). Fatty acid challenges rapidly induce an
insulin resistant state, which is mediated in a number of ways.
These include acylcarnitine accumulation, beta-cell dysfunction,
altered intra-myocellular lipid partitioning and oxidation, reduced
non-oxidative glucose disposal, PKCθ activation and DAG enrich-
ment in skeletal muscle, as well as inducing a postprandial pro-
inflammatory state (Cruz-Teno et al., 2012; Jans et al., 2012a;
Nowotny et al., 2013; Stephens et al., 2014; Wuesten et al., 2005).
However the effects of chronic dietary fat modifications are more
complex and the data are mixed, as reviewed extensively
212 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214
elsewhere (Murphy et al., 2014). Briefly, there is no doubt that
obesity as a result of HFD, particularly if enriched in SFA, promotes
a pro-inflammatory, insulin resistant state characteristic of MetS,
with a greater risk of T2D (Minihane et al., 2015). Indeed recent
mechanistic studies have provided very strong evidence in relation
to molecular processes involved. Nevertheless, even if this para-
digm is true, simply removing dietary SFA, with or without re-
placement with other fatty acid (e.g. MUFA, (n
3 LCPUFA)), does
not necessarily resolve the pro-inflammatory, insulin resistant
state. A recent review highlighted the challenges arising from the
inconsistencies within the scientific evidence to date (Minihane
et al., 2015). All diet and lifestyle intervention studies are chal-
lenged by apparent responders versus non-responders, wherein
typically 30–40% of individuals ‘respond’ to the intervention. This
apparent difference in treatment efficacy goes beyond simple
compliance and may reflect different genetic or phenotypic char-
acteristics. For example, Yubero-Serrano et al. demonstrated that
MetS subjects with insulin resistance, displaying more extensive
metabolic complications, responded more effectively to dietary fat
modification, wherein dietary SFA were reduced by replacement
with MUFA and (n
3 LCPUFA) (Yubero-Serrano et al., 2015).
Conversely, MetS subjects without insulin resistance were more
sensitive to the detrimental effects of dietary SFA intake. Thus the
metabolic phenotype (or metabotypes) of subjects may modulate
response to the quantity and quality of dietary fatty acid on MetS
risk factors; which suggests that targeted or personalized dietary
therapies based on distinct metabotypes may be more effective
(Phillips et al., 2012). This variation in responsiveness is the focus
of current research, as we attempt to address the knowledge gap
with respect to those genotype-phenotype combinations that de-
termine sensitivity to fatty acids; both in terms of identifying risk
and enhancing therapeutic efficacy. Furthermore, systemic bio-
markers of inflammation do not necessarily reflect the potent
biological effects of inflammatory mediators in the metabolic tis-
sues, including adipose tissue, liver and skeletal muscle. Clearly,
there are several gaps in knowledge relating to the inter-re-
lationships between dietary fatty acid, inflammation and meta-
bolism. These need to be resolved given the continued risks arising
from excessive dietary fat intake and escalating obesity rates with
concomitant risk of NALFD, MetS, T2D and CVD.
Acknowledgements
AAC, RMC, CLL and HMR are supported by Science Foundation
Ireland Principal Investigator Programme (11/PI/1119). AMM and
HMR were funded by the Irish Department of Agriculture, Food
and the Marine, NUTRIMAL Programme (14F 882). RMC and AMM
were funded by the National Children's Research Centre, Crumlin,
Ireland (Grant number: B/11/1).
References
Aguirre, V., Werner, E.D., Giraud, J., Lee, Y.H., Shoelson, S.E., White, M.F., 2002.
Phosphorylation of Ser307 in insulin receptor substrate-1 blocks interactions
with the insulin receptor and inhibits insulin action. J. Biol. Chem. 277,
1531–1537. http://dx.doi.org/10.1074/jbc.M101521200.
Alberti, K.G.M.M., Eckel, R.H., Grundy, S.M., Zimmet, P.Z., Cleeman, J.I., Donato, K.A.,
Fruchart, J.-C., James, W.P.T., Loria, C.M., Smith, S.C., 2009. Harmonizing the
metabolic syndrome: a joint interim statement of the International Diabetes
Federation Task Force on Epidemiology and Prevention; National Heart, Lung,
and Blood Institute; American Heart Association; World Heart Federation; In-
ternational Atherosclerosis Society; and International Association for the Study
of Obesity. Circulation 120, 1640–1645. http://dx.doi.org/10.1161/
CIRCULATIONAHA.109.192644.
Arner, P., 1995. Differences in lipolysis between human subcutaneous and omental
adipose tissues. Ann. Med. 27, 435–438. http://dx.doi.org/10.3109/
07853899709002451.
Basu, A., Basu, R., Shah, P., Vella, A., Rizza, R.A., Jensen, M.D., 2001. Systemic and
regional free fatty acid metabolism in type 2 diabetes. Am. J. Physiol. En-
docrinol. Metab. 280, E1000–E1006.
Benhamed, F., Denechaud, P.D., Lemoine, M., Robichon, C., Moldes, M., Bertrand-
Michel, J., Ratziu, V., Serfaty, L., Housset, C., Capeau, J., Girard, J., Guillou, H.,
Postic, C., 2012. The lipogenic transcription factor ChREBP dissociates hepatic
steatosis from insulin resistance in mice and humans. J. Clin. Investig. 122,
2176–2194. http://dx.doi.org/10.1172/JCI41636.
Carey, A.L., Steinberg, G.R., Macaulay, S.L., Thomas, W.G., Holmes, A.G., Ramm, G.,
Prelovsek, O., Hohnen-Behrens, C., Watt, M.J., James, D.E., Kemp, B.E., Pedersen,
B.K., Febbraio, M.A., 2006. Interleukin-6 increases insulin-stimulated glucose
disposal in humans and glucose uptake and fatty acid oxidation in vitro via
AMP-activated protein kinase. Diabetes 55, 2688–2697. http://dx.doi.org/
10.2337/db05-1404.
Chawla, A., Nguyen, K.D., Goh, Y.P.S., 2011. Macrophage-mediated inflammation in
metabolic disease. Nat. Rev. Immunol. 11, 738–749. http://dx.doi.org/10.1038/
nri3071.
Connaughton, R.M., McMorrow, A.M., McGillicuddy, F.C., Lithander, F.E., Roche, H.
M., 2016. Impact of anti-inflammatory nutrients on obesity-associated meta-
bolic-inflammation from childhood through to adulthood. Proc. Nutr. Soc.,
1–10. http://dx.doi.org/10.1017/S0029665116000070.
Cruz-Teno, C., Pérez-Martínez, P., Delgado-Lista, J., Yubero-Serrano, E.M., García-
Ríos, A., Marín, C., Gómez, P., Jiménez-Gómez, Y., Camargo, A., Rodríguez-Can-
talejo, F., Malagón, M.M., Pérez-Jiménez, F., Roche, H.M., López-Miranda, J.,
2012. Dietary fat modifies the postprandial inflammatory state in subjects with
metabolic syndrome: the LIPGENE study. Mol. Nutr. Food Res. 56, 854–865.
http://dx.doi.org/10.1002/mnfr.201200096.
De Roos, B., Rungapamestry, V., Ross, K., Rucklidge, G., Reid, M., Duncan, G., Horgan,
G., Toomey, S., Browne, J., Loscher, C.E., Mills, K.H.G., Roche, H.M., 2009. At-
tenuation of inflammation and cellular stress-related pathways maintains in-
sulin sensitivity in obese type I interleukin-1 receptor knockout mice on a high-
fat diet. Proteomics 9, 3244–3256.
Després, J.-P., Lemieux, I., 2006. Abdominal obesity and metabolic syndrome. Nat-
ure 444, 881–887. http://dx.doi.org/10.1038/nature05488.
Dinarello, C.A., 2009. Immunological and inflammatory functions of the inter-
leukin-1 family. Annu. Rev. Immunol. 27, 519–550. http://dx.doi.org/10.1146/
annurev.immunol.021908.132612.
Eckel, R.H., Alberti, K., Grundy, S.M., Zimmet, P.Z., 2010. The metabolic syndrome.
Lancet 375, 181–183. http://dx.doi.org/10.1016/S0140-6736(09)61794-3.
Eckel, R.H., Grundy, S.M., Zimmet, P.Z., 2005. The metabolic syndrome. Lancet 365,
1415–1428. http://dx.doi.org/10.1016/S0140-6736(05)66378-7.
Emanuelli, B., Peraldi, P., Filloux, C., Sawka-Verhelle, D., Hilton, D., Van Obberghen,
E., 2000. SOCS-3 is an insulin-induced negative regulator of insulin signaling. J.
Biol. Chem. 275, 15985–15991.
Fasshauer, M., Klein, J., Lossner, U., Paschke, R., 2003. Interleukin (IL)
6 mRNA
expression is stimulated by insulin, isoproterenol, tumour necrosis factor alpha,
growth hormone, and IL-6 in 3T3-L1 adipocytes. Horm. Metab. Res. 35,
147–152. http://dx.doi.org/10.1055/s-2003-39075.
Fearon, K.C.H., Glass, D.J., Guttridge, D.C., 2012. Cancer cachexia: mediators, sig-
naling, and metabolic pathways. Cell Metab. 16, 153–166. http://dx.doi.org/
10.1016/j.cmet.2012.06.011.
Feuerer, M., Herrero, L., Cipolletta, D., Naaz, A., Wong, J., Nayer, A., Lee, J., Goldfine,
A.B., Benoist, C., Shoelson, S., Mathis, D., 2009. Lean, but not obese, fat is en-
riched for a unique population of regulatory T cells that affect metabolic
parameters. Nat. Med. 15, 930–939. http://dx.doi.org/10.1038/nm.2002.
Finucane, O.M., Lyons, C.L., Murphy, A.M., Reynolds, C.M., Klinger, R., Healy, N.P.,
Cooke, A.A., Coll, R.C., McAllan, L., Nilaweera, K.N., O’Reilly, M.E., Tierney, A.C.,
Morine, M.J., Alcala-Diaz, J.F., Lopez-Miranda, J., O’Connor, D.P., O’Neill, L.A.,
McGillicuddy, F.C., Roche, H.M., 2015. Monounsaturated fatty acid–enriched
high-fat diets impede adipose NLRP3 inflammasome–mediated IL-1β secretion
and insulin resistance despite obesity. Diabetes . http://dx.doi.org/10.2337/
db14-1098, db141098.
Galbo, T., Perry, R.J., Jurczak, M.J., Camporez, J.-P.G., Alves, T.C., Kahn, M., Guigni, B.
A., Serr, J., Zhang, D., Bhanot, S., Samuel, V.T., Shulman, G.I., 2013. Saturated and
unsaturated fat induce hepatic insulin resistance independently of TLR-4 sig-
naling and ceramide synthesis in vivo. Proc. Natl. Acad. Sci. USA 110,
12780–12785. http://dx.doi.org/10.1073/pnas.1311176110.
Gao, Z., Hwang, D., Bataille, F., Lefevre, M., York, D., Quon, M.J., Ye, J., 2002. Serine
phosphorylation of insulin receptor substrate 1 by inhibitor kappa B kinase
complex. J. Biol. Chem. 277, 48115–48121. http://dx.doi.org/10.1074/jbc.
M209459200.
Glass, C.K., Olefsky, J.M., 2012. Inflammation and lipid signaling in the etiology of
insulin resistance. Cell Metab. 15, 635–645. http://dx.doi.org/10.1016/j.
cmet.2012.04.001.
He, J., Usui, I., Ishizuka, K., Kanatani, Y., Hiratani, K., Iwata, M., Bukhari, A., Haruta, T.,
Sasaoka, T., Kobayashi, M., 2006. Interleukin-1alpha inhibits insulin signaling
with phosphorylating insulin receptor substrate-1 on serine residues in 3T3-L1
adipocytes. Mol. Endocrinol. 20, 114–124. http://dx.doi.org/10.1210/me.2005–
0107.
Herman, M.A., Peroni, O.D., Villoria, J., Schön, M.R., Abumrad, N.A., Blüher, M., Klein,
S., Kahn, B.B., 2012. A novel ChREBP isoform in adipose tissue regulates sys-
temic glucose metabolism. Nature 484, 333–338. http://dx.doi.org/10.1038/
nature10986.
Hirosumi, J., Tuncman, G., Chang, L., Görgün, C.Z., Uysal, K.T., Maeda, K., Karin, M.,
Hotamisligil, G.S., 2002. A central role for JNK in obesity and insulin resistance.
Nature 420, 333–336. http://dx.doi.org/10.1038/nature01137.
 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214
Holland, W.L., Bikman, B.T., Wang, L., Yuguang, G., Sargent, K.M., Bulchand, S.,
Knotts, T.A., Shui, G., Clegg, D.J., Wenk, M.R., Pagliassotti, M.J., Scherer, P.E.,
Summers, S.A., 2011. Lipid-induced insulin resistance mediated by the proin-
flammatory receptor TLR4 requires saturated fatty acid – induced ceramide
biosynthesis in mice. J. Clin. Investig., 121. http://dx.doi.org/10.1172/
JCI43378.1858.
Hong, E.-G., Ko, H.J., Cho, Y.-R., Kim, H.-J., Ma, Z., Yu, T.Y., Friedline, R.H., Kurt-Jones,
E., Finberg, R., Fischer, M.A., Granger, E.L., Norbury, C.C., Hauschka, S.D., Phil-
brick, W.M., Lee, C.-G., Elias, J.A., Kim, J.K., 2009. Interleukin-10 prevents diet-
induced insulin resistance by attenuating macrophage and cytokine response in
skeletal muscle. Diabetes 58, 2525–2535. http://dx.doi.org/10.2337/db08-1261.
Hong, H.C., Hwang, S.Y., Ryu, J.Y., Yoo, H.J., Seo, J.-A., Kim, S.G., Kim, N.H., Baik, S.H.,
Choi, D.S., Choi, K.M., 2015. The synergistic impact of nonalcoholic fatty liver
disease and metabolic syndrome on subclinical atherosclerosis. Clin. En-
docrinol. . http://dx.doi.org/10.1111/cen.12940
Hotamisligil, G.S., 2006. Inflammation and metabolic disorders. Nature 444,
860–867. http://dx.doi.org/10.1038/nature05485.
Hotamisligil, G.S., Peraldi, P., Budavari, A., Ellis, R., White, M.F., Spiegelman, B.M.,
1996. IRS-1-mediated inhibition of insulin receptor tyrosine kinase activity in
TNF-alpha- and obesity-induced insulin resistance. Science 271, 665–668.
Hotamisligil, G.S., Shargill, N.S., Spiegelman, B.M., 1993. Adipose expression of tu-
mor necrosis factor-alpha: direct role in obesity-linked insulin resistance. Sci-
ence 259, 87–91 (80).
Jager, J., Grémeaux, T., Cormont, M., Le Marchand-Brustel, Y., Tanti, J.-F., 2007. In-
terleukin-1beta-induced insulin resistance in adipocytes through down-reg-
ulation of insulin receptor substrate-1 expression. Endocrinology 148, 241–251.
http://dx.doi.org/10.1210/en.2006–0692.
Jans, A., Konings, E., Goossens, G.H., Bouwman, F.G., Moors, C.C., Boekschoten, M.V.,
Afman, L.A., Müller, M., Mariman, E.C., Blaak, E.E., 2012a. PUFAs acutely affect
triacylglycerol-derived skeletal muscle fatty acid uptake and increase post-
prandial insulin sensitivity. Am. J. Clin. Nutr. 95, 825–836. http://dx.doi.org/
10.3945/ajcn.111.028787.
Jans, A., Van Hees, A.M.J., Gjelstad, I.M.F., Sparks, L.M., Tierney, A.C., Ris??rus, U.,
Drevon, C.A., Schrauwen, P., Roche, H.M., Blaak, E.E., 2012b. Impact of dietary fat
quantity and quality on skeletal muscle fatty acid metabolism in subjects with
the metabolic syndrome. Metabolism 61, 1554–1565. http://dx.doi.org/10.1016/
j.metabol.2012.04.003.
Jo, J., Gavrilova, O., Pack, S., Jou, W., Mullen, S., Sumner, A.E., Cushman, S.W., Periwal,
V., 2009. Hypertrophy and/or hyperplasia : dynamics of adipose tissue growth.
PLoS Comput. Biol., 5. http://dx.doi.org/10.1371/journal.pcbi.1000324.
Johnson, A.M.F., Olefsky, J.M., 2013. The origins and drivers of insulin resistance.
Cell 152, 673–684. http://dx.doi.org/10.1016/j.cell.2013.01.041.
Jornayvaz, F.R., Shulman, G.I., 2012. Diacylglycerol activation of protein kinase Cε
and hepatic insulin resistance. Cell Metab. 15, 574–584. http://dx.doi.org/
10.1016/j.cmet.2012.03.005.
Kamei, N., Tobe, K., Suzuki, R., Ohsugi, M., Watanabe, T., Kubota, N., Ohtsuka-Ko-
watari, N., Kumagai, K., Sakamoto, K., Kobayashi, M., Yamauchi, T., Ueki, K.,
Oishi, Y., Nishimura, S., Manabe, I., Hashimoto, H., Ohnishi, Y., Ogata, H., To-
kuyama, K., Tsunoda, M., Ide, T., Murakami, K., Nagai, R., Kadowaki, T., 2006.
Overexpression of monocyte chemoattractant protein-1 in adipose tissues
causes macrophage recruitment and insulin resistance. J. Biol. Chem. 281,
26602–26614. http://dx.doi.org/10.1074/jbc.M601284200.
Kim, J.K., Gavrilova, O., Chen, Y., Reitman, M.L., Shulman, G.I., 2000. Mechanism of
insulin resistance in A-ZIP/F-1 fatless mice. J. Biol. Chem. 275, 8456–8460. http:
//dx.doi.org/10.1074/jbc.275.12.8456.
Koenen, T.B., Stienstra, R., Van Tits, L.J., Joosten, L.A.B., Van Velzen, J.F., Hijmans, A.,
Pol, J.A., Van Der Vliet, J.A., Netea, M.G., Tack, C.J., Stalenhoef, A.F.H., De Graaf, J.,
2011. The inflammasome and caspase-1 activation: a new mechanism under-
lying increased inflammatory activity in human visceral adipose tissue. En-
docrinology 152, 3769–3778. http://dx.doi.org/10.1210/en.2010–1480.
Kylin, E., 1923. Studien über das Hypertonie-Hyperglykämie-Hyperurikämie Syn-
drom. 44. Zentralblatt für Inn. Medizin, pp. 105–112.
Lafontan, M., Langin, D., 2009. Lipolysis and lipid mobilization in human adipose
tissue. Prog. Lipid Res. 48, 275–297. http://dx.doi.org/10.1016/j.
plipres.2009.05.001.
Lewis, G., O’meara, N., 1990. Postprandial lipoprotein metabolism in normal and
obese subjects: comparison after the vitamin A fat-loading test*. J. Clin. En-
docrinol. Metab., 71.
Liu, J., Divoux, A., Sun, J., Zhang, J., Clément, K., Glickman, J.N., Sukhova, G.K.,
Wolters, P.J., Du, J., Gorgun, C.Z., Doria, A., Libby, P., Blumberg, R.S., Kahn, B.B.,
Hotamisligil, G.S., Shi, G.-P., 2009. Genetic deficiency and pharmacological
stabilization of mast cells reduce diet-induced obesity and diabetes in mice.
Nat. Med. 15, 940–945. http://dx.doi.org/10.1038/nm.1994.
Lumeng, C.N., DelProposto, J.B., Westcott, D.J., Saltiel, A.R., 2008. Phenotypic
switching of adipose tissue macrophages with obesity is generated by spatio-
temporal differences in macrophage subtypes. Diabetes 57, 3239–3246. http:
//dx.doi.org/10.2337/db08-0872.
Lundgren, M., Svensson, M., Lindmark, S., 2007. Fat cell enlargement is an in-
dependent marker of insulin resistance and “hyperleptinaemia. Diabetologia.
Martinon, F., Burns, K., Tschopp, J., 2002. The inflammasome: a molecular platform
triggering activation of inflammatory caspases and processing of proIL-beta.
Mol. Cell 10, 417–426.
McGillicuddy, F.C., Harford, K.A., Reynolds, C.M., Oliver, E., Claessens, M., Mills, K.H.
G., Roche, H.M., 2011. Lack of interleukin-1 receptor I (IL-1RI) protects mice
from high-fat diet-induced adipose tissue inflammation coincident with im-
proved glucose homeostasis. Diabetes 60, 1688–1698. http://dx.doi.org/
213
10.2337/db10-1278.
Minihane, A.M., Vinoy, S., Russell, W.R., Baka, A., Roche, H.M., Tuohy, K.M., Teeling, J.
L., Blaak, E.E., Fenech, M., Vauzour, D., McArdle, H.J., Kremer, B.H.A., Sterkman,
L., Vafeiadou, K., Benedetti, M.M., Williams, C.M., Calder, P.C., 2015. Low-grade
inflammation, diet composition and health: current research evidence and its
translation. Br. J. Nutr. 114, 999–1012. http://dx.doi.org/10.1017/
S0007114515002093.
Munoz-Canoves, P., Scheele, C., Pedersen, B.K., Serrano, A.L., 2013. Interleukin-6
myokine signaling in skeletal muscle: a double-edged sword? FEBS J. 280,
4131–4148. http://dx.doi.org/10.1111/febs.12338.
Muoio, D.M., Neufer, P.D., 2012. Lipid-Induced mitochondrial stress and insulin
action in muscle. Cell Metab. 15, 595–605. http://dx.doi.org/10.1016/j.
cmet.2012.04.010.
Murphy, A.M., Lyons, C.L., Finucane, O.M., Roche, H.M., 2014. Interactions between
differential fatty acids and inflammatory stressors-impact on metabolic health.
Prostaglandins Leukot. Essent. Fat. Acids 92, 49–55. http://dx.doi.org/10.1016/j.
plefa.2014.05.003.
Nagle, C.A., An, J., Shiota, M., Torres, T.P., Cline, G.W., Liu, Z.X., Wang, S., Catlin, R.L.,
Shulman, G.I., Newgard, C.B., Coleman, R.A., 2007. Hepatic overexpression of
glycerol-sn-3-phosphate acyltransferase 1 in rats causes insulin resistance. J.
Biol. Chem. 282, 14807–14815. http://dx.doi.org/10.1074/jbc.M611550200.
Nakamura, T., Furuhashi, M., Li, P., Cao, H., Tuncman, G., Sonenberg, N., Gorgun, C.Z.,
Hotamisligil, G.S., 2010. Double-stranded RNA-dependent protein kinase links
pathogen sensing with stress and metabolic homeostasis. Cell 140, 338–348.
http://dx.doi.org/10.1016/j.cell.2010.01.001.
Nedergaard, J., Bengtsson, T., Cannon, B., 2007. Unexpected evidence for active
brown adipose tissue in adult humans. Am. J. Physiol. Endocrinol. Metab. 293,
E444–E452. http://dx.doi.org/10.1152/ajpendo.00691.2006.
Niemand, C., Nimmesgern, A., Haan, S., Fischer, P., Schaper, F., Rossaint, R., Heinrich,
P.C., Müller-Newen, G., 2003. Activation of STAT3 by IL-6 and IL-10 in primary
human macrophages is differentially modulated by suppressor of cytokine
signaling 3. J. Immunol. 170, 3263–3272.
Nishimura, S., Manabe, I., Nagasaki, M., Eto, K., Yamashita, H., Ohsugi, M., Otsu, M.,
Hara, K., Ueki, K., Sugiura, S., Yoshimura, K., Kadowaki, T., Nagai, R., 2009.
CD8þ effector T cells contribute to macrophage recruitment and adipose tissue
inflammation in obesity. Nat. Med. 15, 914–920. http://dx.doi.org/10.1038/
nm.1964.
Nowotny, B., Zahiragic, L., Krog, D., Nowotny, P.J., Herder, C., Carstensen, M., Yosh-
imura, T., Szendroedi, J., Phielix, E., Schadewaldt, P., Schloot, N.C., Shulman, G.I.,
Roden, M., 2013. Mechanisms underlying the onset of oral lipid – induced
skeletal muscle insulin resistance in humans. Diabetes 62, 2240–2248. http:
//dx.doi.org/10.2337/db12-1179.
Odegaard, J.I., Chawla, A., 2013. Pleiotropic actions of insulin resistance and in-
flammation in metabolic homeostasis. Science 339, 172–177. http://dx.doi.org/
10.1126/science.1230721.
Oh, D.Y., Talukdar, S., Bae, E.J., Imamura, T., Morinaga, H., Fan, W., Li, P., Lu, W.J.,
Watkins, S.M., Olefsky, J.M., 2010. GPR120 is an omega-3 fatty acid receptor
mediating potent anti-inflammatory and insulin-sensitizing effects. Cell 142,
687–698. http://dx.doi.org/10.1016/j.cell.2010.07.041.
Oh, D.Y., Walenta, E., Akiyama, T.E., Lagakos, W.S., Lackey, D., Pessentheiner, A.R.,
Sasik, R., Hah, N., Chi, T.J., Cox, J.M., Powels, M.A., Di Salvo, J., Sinz, C., Watkins, S.
M., Armando, A.M., Chung, H., Evans, R.M., Quehenberger, O., McNelis, J.,
Bogner-Strauss, J.G., Olefsky, J.M., 2014. A Gpr120-selective agonist improves
insulin resistance and chronic inflammation in obese mice. Nat. Med. 20,
942–947. http://dx.doi.org/10.1038/nm.3614.
Oliver, E., McGillicuddy, F.C., Harford, K.A., Reynolds, C.M., Phillips, C.M., Ferguson, J.
F., Roche, H.M., 2012. Docosahexaenoic acid attenuates macrophage-induced
inflammation and improves insulin sensitivity in adipocytes-specific differ-
ential effects between LC n
3 PUFA. J. Nutr. Biochem. 23, 1192–1200. http://dx.
doi.org/10.1016/j.jnutbio.2011.06.014.
Osborn, O., Olefsky, J.M., 2012. The cellular and signaling networks linking the
immune system and metabolism in disease. Nat. Med. 18, 363–374. http://dx.
doi.org/10.1038/nm.2627.
Pedersen, B.K., Febbraio, M.A., 2007. Point: Interleukin-6 does have a beneficial role
in insulin sensitivity and glucose homeostasis. J. Appl. Physiol. 102, 814–816.
http://dx.doi.org/10.1152/japplphysiol.01208.2006.
Phillips, C.M., Tierney, A.C., Perez-Martinez, P., Defoort, C., Blaak, E.E., Gjelstad, I.M.
F., Lopez-Miranda, J., Kiec-Klimczak, M., Malczewska-Malec, M., Drevon, C.A.,
Hall, W., Lovegrove, J.A., Karlstrom, B., Risérus, U., Roche, H.M., 2012. Obesity
and body fat classification in the metabolic syndrome: impact on cardiometa-
bolic risk metabotype. Obesity 21, 154–161. http://dx.doi.org/10.1038/
oby.2012.188.
Phillips, C.M., Tierney, A.C., Roche, H.M., 2008. Gene-nutrient interactions in the
metabolic syndrome. J. Nutr. Nutr. 1, 136–151. http://dx.doi.org/10.1159/
000112461.
Powell, D.J., Turban, S., Gray, A., Hajduch, E., Hundal, H.S., 2004. Intracellular cer-
amide synthesis and protein kinase Czeta activation play an essential role in
palmitate-induced insulin resistance in rat L6 skeletal muscle cells. Biochem. J.
382, 619–629. http://dx.doi.org/10.1042/BJ20040139.
Reaven, G.M., 1988. Role of insulin resistance in human disease. Diabetes 37,
1595–1607. http://dx.doi.org/10.2337/diab.37.12.1595.
Reynolds, C.M., Mcgillicuddy, F.C., Harford, K.A., Finucane, O.M., Mills, K.H.G., Roche,
H.M., 2012. Dietary saturated fatty acids prime the NLRP3 inflammasome via
TLR4 in dendritic cells-implications for diet-induced insulin resistance. Mol.
Nutr. Food Res. 56, 1212–1222. http://dx.doi.org/10.1002/mnfr.201200058.
Rosenwald, M., Wolfrum, C., 2014. The origin and definition of brite versus white
214 A.A. Cooke et al. / European Journal of Pharmacology 785 (2016) 207–214
and classical brown adipocytes. Adipocyte.
Rosqvist, F., Iggman, D., Kullberg, J., Cedernaes, J., Johansson, H.E., Larsson, A., Jo-
hansson, L., Ahlström, H., Arner, P., Dahlman, I., Risérus, U., 2014. Overfeeding
polyunsaturated and saturated fat causes distinct effects on liver and visceral
fat accumulation in humans. Diabetes 63, 2356–2368. http://dx.doi.org/
10.2337/db13-1622.
Rotter, V., Nagaev, I., Smith, U., 2003. Interleukin-6 (IL-6) induces insulin resistance
in 3T3-L1 adipocytes and is, like IL-8 and tumor necrosis factor-, overexpressed
in human fat cells from insulin-resistant subjects. J. Biol. Chem. 278,
45777–45784. http://dx.doi.org/10.1074/jbc.M301977200.
Samuel, V.T., Shulman, G.I., 2012. Mechanisms for insulin resistance: common
threads and missing links. Cell 148, 852–871. http://dx.doi.org/10.1016/j.
cell.2012.02.017.
Saraiva, M., O’Garra, A., 2010. The regulation of IL-10 production by immune cells.
Nat. Rev. Immunol. 10, 170–181. http://dx.doi.org/10.1038/nri2711.
Schipper, H.S., Prakken, B., Kalkhoven, E., Boes, M., 2012. Adipose tissue-resident
immune cells: key players in immunometabolism. Trends Endocrinol. Metab.
23, 407–415. http://dx.doi.org/10.1016/j.tem.2012.05.011.
Schrauwen, P., Timmers, S., 2014. Can resveratrol help to maintain metabolic
health? Proc. Nutr. Soc. 73, 271–277. http://dx.doi.org/10.1017/
S0029665113003856.
Schweiger, M., Schreiber, R., 2006. Adipose triglyceride lipase and hormone-sen-
sitive lipase are the major enzymes in adipose tissue triacylglycerol catabolism.
J. Biol.
Shames, B.D., Selzman, C.H., Meldrum, D.R., Pulido, E.J., Barton, H.A., Meng, X.,
Harken, A.H., McIntyre, R.C., 1998. Interleukin-10 stabilizes inhibitory kappaB-
alpha in human monocytes. Shock 10, 389–394.
Stephens, F., Mendis, B., Shannon, C., Cooper, S., Ortori, C., Barrett, D., Mansell, P.,
Tsintzas, K., 2014. Fish oil omega-3 fatty acids partially prevent lipid-induced
insulin resistance in human skeletal muscle without limiting acylcarnitine ac-
cumulation. Clin. Sci. 127, 315–322.
Sun, K., Tordjman, J., Clément, K., Scherer, P.E., 2013. Fibrosis and adipose tissue
dysfunction. Cell Metab. 18, 470–477. http://dx.doi.org/10.1016/j.
cmet.2013.06.016.
Sung, K.-C., Wild, S.H., Kwag, H.J., Byrne, C.D., 2012. Fatty liver, insulin resistance,
and features of metabolic syndrome: relationships with coronary artery cal-
cium in 10,153 people. Diabetes Care 35, 2359–2364. http://dx.doi.org/10.2337/
dc12-0515.
Targher, G., 2007. Non-alcoholic fatty liver disease, the metabolic syndrome and the
risk of cardiovascular disease: the plot thickens. Diabet. Med. 24, 1–6. http://dx.
doi.org/10.1111/j.1464–5491.2007.02025.x.
Targher, G., Arcaro, G., 2007. Non-alcoholic fatty liver disease and increased risk of
cardiovascular disease. Atherosclerosis 191, 235–240. http://dx.doi.org/10.1016/
j.atherosclerosis.2006.08.021.
Targher, G., Bertolini, L., Padovani, R., Poli, F., Scala, L., Tessari, R., Zenari, L., Falezza,
G., 2006. Increased Prevalence of Cardiovascular Disease in Type 2 Diabetic
Patients With Non-Alcoholic Fatty Liver Disease, pp. 403–409. doi: 〈http://dx.
doi.org/10.1111/j.1464-5491.2005.01817.x〉.
Tchkonia, T., Tchoukalova, Y.D., Giorgadze, N., Pirtskhalava, T., Karagiannides, I.,
Forse, R.A., Koo, A., Stevenson, M., Chinnappan, D., Cartwright, A., Jensen, M.D.,
Kirkland, J.L., Abun-, J.L.K., 2005. Abundance of Two Human Preadipocyte
Subtypes With Distinct Capacities for Replication , Adipogenesis , and Apoptosis
Varies Among Fat Depots. Vol. 02118, pp. 267–277. doi: 〈http://dx.doi.org/10.
1152/ajpendo.00265.2004〉.
Tschopp, J., Schroder, K., 2010. NLRP3 inflammasome activation: the convergence of
multiple signalling pathways on ROS production? Nat. Rev. Immunol. 10,
210–215. http://dx.doi.org/10.1038/nri2725.
Tumova, J., Andel, M., Trnka, J., 2015. Excess of free fatty acids as a cause of me-
tabolic dysfunction in skeletal muscle. Physiol. Res.
Ueki, K., Kondo, T., Kahn, C.R., 2004. Suppressor of cytokine signaling 1 ( SOCS-1 )
and SOCS-3 cause insulin resistance through inhibition of tyrosine
phosphorylation of insulin receptor substrate proteins by discrete mechanisms.
Mol. Cell. Biol. 24, 5434–5446. http://dx.doi.org/10.1128/MCB.24.12.5434.
Uysal, K.T., Wiesbrock, S.M., Marino, M.W., Hotamisligil, G.S., 1997. Protection from
obesity-induced insulin resistance in mice lacking TNF-alpha function. Nature
389, 610–614. http://dx.doi.org/10.1038/39335.
Vague, J., 1947. La différentiation sexuelle, facteur déterinant des formes de l’obé-
sité. Presse Méd. 55, 339–340.
Virtue, S., Vidal-Puig, A., 2010. Adipose tissue expandability, lipotoxicity and the
Metabolic Syndrome–an allostatic perspective. Biochim. Biophys. Acta 1801,
338–349. http://dx.doi.org/10.1016/j.bbalip.2009.12.006.
Virtue, S., Vidal-Puig, A., 2008. It's not how fat you are, it's what you do with it that
counts. PLoS Biol. 6. http://dx.doi.org/10.1371/journal.pbio.0060237.
Weisberg, S.P., McCann, D., Desai, M., Rosenbaum, M., Leibel, R.L., Ferrante, A.W.,
2003. Obesity is associated with macrophage accumulation in adipose tissue. J.
Clin. Investig. 112, 1796–1808. http://dx.doi.org/10.1172/JCI19246.
Winer, D.A., Winer, S., Shen, L., Wadia, P.P., Yantha, J., Paltser, G., Tsui, H., Wu, P.,
Davidson, M.G., Alonso, M.N., Leong, H.X., Glassford, A., Caimol, M., Kenkel, J.A.,
Tedder, T.F., McLaughlin, T., Miklos, D.B., Dosch, H.-M., Engleman, E.G., 2011. B
cells promote insulin resistance through modulation of T cells and production
of pathogenic IgG antibodies. Nat. Med. 17, 610–617. http://dx.doi.org/10.1038/
nm.2353.
Wolsk, E., Mygind, H., Grøndahl, T.S., Pedersen, B.K., Hall, G.VAN., 2010. IL-6 se-
lectively stimulates fat metabolism in human skeletal muscle. Am. J. Physiol.
Endocrinol. Metab. 299, E832–E840. http://dx.doi.org/10.1152/
ajpendo.00328.2010.
Wu, D., Molofsky, A.B., Liang, H.-E., Ricardo-Gonzalez, R.R., Jouihan, H.A., Bando, J.K.,
Chawla, A., Locksley, R.M., 2011. Eosinophils sustain adipose alternatively ac-
tivated macrophages associated with glucose homeostasis. Science 332,
243–247. http://dx.doi.org/10.1126/science.1201475 (80).
Wuesten, O., Balz, C.H., Bretzel, R.G., Kloer, H.-U., Hardt, P.D., 2005. Effects of oral fat
load on insulin output and glucose tolerance in healthy control subjects and
obese patients without diabetes. Diabetes Care 28, 360–365. http://dx.doi.org/
10.2337/diacare.28.2.360.
Xiao, C., Giacca, A., Carpentier, A., Lewis, G.F., 2006. Differential effects of mono-
unsaturated, polyunsaturated and saturated fat ingestion on glucose-stimu-
lated insulin secretion, sensitivity and clearance in overweight and obese, non-
diabetic humans. Diabetologia 49, 1371–1379. http://dx.doi.org/10.1007/
s00125-006-0211-x.
Xu, H., Barnes, G.T., Yang, Q., Tan, G., Yang, D., Chou, C.J., Sole, J., Nichols, A., Ross, J.S.,
Tartaglia, L.A., Chen, H., 2003. Chronic inflammation in fat plays a crucial role in
the development of obesity-related insulin resistance. J. Clin. Investig. 112,
1821–1830. http://dx.doi.org/10.1172/JCI19451.
Yore, M.M., Syed, I., Moraes-vieira, P.M., Zhang, T., Herman, M.A., Homan, E.A., Patel,
R.T., Lee, J., Chen, S., Peroni, O.D., Dhaneshwar, A.S., Hammarstedt, A., Smith, U.,
Mcgraw, T.E., Saghatelian, A., Kahn, B.B., 2014. Discovery of a class of en-
dogenous mammalian lipids with anti-diabetic and anti-inflammatory effects.
Cell 159, 318–332. http://dx.doi.org/10.1016/j.cell.2014.09.035.
Yu, Y., Ginsberg, H., 2005. Adipocyte signaling and lipid homeostasis sequelae of
insulin-resistant adipose tissue. Circ. Res.
Yuan, M., Konstantopoulos, N., Lee, J., Hansen, L., Li, Z.W., Karin, M., Shoelson, S.E.,
2001. Reversal of obesity- and diet-induced insulin resistance with salicylates
or targeted disruption of Ikkbeta. Science 293, 1673–1677. http://dx.doi.org/
10.1126/science.1061620.
Yubero-Serrano, E.M., Delgado-Lista, J., Tierney, A.C., Perez-Martinez, P., Garcia-
Rios, A., Alcala-Diaz, J.F., Castano, J.P., Tinahones, F.J., Drevon, C.A., Defoort, C.,
Blaak, E.E., Dembinska-Kiec, A., Riserus, U., Lovegrove, J.A., Perez-Jimenez, F.,
Roche, H.M., Lopez-Miranda, J., 2015. Insulin resistance determines a differ-
ential response to changes in dietary fat modification on metabolic syndrome
risk factors: the LIPGENE study. Am. J. Clin. Nutr., 1509–1517. http://dx.doi.org/
10.3945/ajcn.115.111286.