Journal of Functional Foods 106 (2023) 105600

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Journal of Functional Foods

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

Supplementation with CO induces lipogenesis in adipose tissue, leptin and

insulin resistance in healthy Swiss mice
Alana Carolina Costa Veras a, Larissa da Silva Bruzasco a, Ana Beatriz Profiro Lopes a,
Beatriz da Silva Franco c, Alessandro Spencer de Souza Holanda c, Andrea Maculano Esteves c,
Marciane Milanski a, b, Adriana Souza Torsoni a, b, Leticia Martins Ignacio-Souza a, b,
Marcio Alberto Torsoni a, b, *
a
Laboratory of Metabolic Disorders, School of Applied Sciences, University of Campinas, Brazil
b
Obesity and Comorbidities Research Center, University of Campinas, Brazil
c
Laboratory of Sleep and Exercise, School of Applied Sciences, University of Campinas, Brazil

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

Keywords: The consumption of saturated fatty acids (SFA), the main compound in coconut oil (CO), can promote insulin and
Coconut oil leptin resistance and are associated with inflammation and obesity. We investigated the effects of CO supple­
Leptin resistance mentation on leptin signaling in healthy mice. Swiss male mice received oral supplementation for eight weeks
Hypothalamus
with 300 μL of water for the control group (CV) or CO (100 or 300 μL). Sensitivity to leptin/insulin was evaluated
Adipose tissue
after eight weeks of supplementation. The CO induced endoplasmic reticulum stress and leptin resistance in the
hypothalamus, as demonstrated by reduced effect on the energy expenditure, hypothalamic pJAK2 and pSTAT3,
and POMC expression. In the adipose tissue, lipogenesis was favored and STAT3 and JAK2 signaling was
impaired after CO supplementation. Furthermore, the supplementation with CO reduced pAKT in the hypo­
thalamus, liver and white adipose tissue. These results show that CO induces hypothalamic and peripheral
resistance to leptin and insulin in healthy mice.

1. Introduction Yanagita, 2010), this is because MCFAs are absorbed in the intestine and
are transported through the portal system directly to the liver. More­
Saturated fatty acid (SFA) is the main component of a fat diet (Krauss over, MCFAs do not depend on the presence of carnitine
and Kris-Etherton, 2020; Harrison et al., 2020; Vandevijvere et al., 2015; palmitoyltransferase-1 (CPT-1) to enter the mitochondrial (Friedman
German and Dillard, 2004) and rich diet in SFA can promote insulin and et al., 1990). In addition, studies have associated benefits of virgin CO
leptin resistance in peripheral and hypothalamic tissues (Lieu et al., with antioxidant properties of phenolics compounds and its beneficial
2021). Leptin resistance may be linked to a failure in leptin transport or role in both prevention and treatment of metabolic syndrome (Nagao
deficits in intracellular signaling mechanisms downstream of leptin and Yanagita, 2010).
(Yazdi et al., 2015). Simultaneously to the development of leptin resis­ On the other hand, SFA stimulates an inflammatory response via
tance, white adipose tissue also develops local resistance to leptin, since TLR2 and TLR4 signaling pathways (Huang et al., 2012; Hwang et al.,
decreased STAT3 activation is observed in adipose tissue of diet-induced 2016). Accordingly, in a previous study we showed that CO consump­
obese animals in basal conditions and stimulated by leptin (Wang et al., tion for eight weeks led to weight gain, higher percentage of fat, reduced
2000). Besides, the peripheral and central signaling of the insulin also energy expenditure, activated inflammatory pathways, both centrally
have an important role in the control of whole-body glucose and energy and peripherally, and triggered an anxious behavior in healthy Swiss
metabolism (Obici et al., 2002; Obici et al., 2002). mice, suggesting the deleterious effect on the body homeostasis (Veras
SFAs, especially medium chain saturated fatty acids (MCFAs), are the et al., 2021). Thus, to evaluate if the CO supplementation also impaired
main compound in coconut oil (CO) (Rahim et al., 2017). Studies indi­ hormonal signaling, we investigated the capacity of leptin to modulate
cate that SFA from coconut oil can play a beneficial role (Nagao and the behavior and energy homeostasis in healthy Swiss mice.

* Corresponding author at: School of Applied Sciences, University of Campinas/Unicamp, Rua Pedro Zaccaria, 1300-Jd. Sta Luiza 13484-350 Limeira, SP, Brazil.
E-mail address: [email protected] (M. Alberto Torsoni).

https://doi.org/10.1016/j.jff.2023.105600
Received 20 January 2023; Received in revised form 24 May 2023; Accepted 26 May 2023
Available online 4 June 2023
1756-4646/© 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
A. Carolina Costa Veras et al. Journal of Functional Foods 106 (2023) 105600

Fig. 1. Reticulum Stress parameters in the hypothalamus of mice supplemented, for 8 weeks, with coconut oil (CO100 or CO300) or water (CV). (A-F)
Western-blotting analysis of Reticulum stress pathways markers, in the hypothalamus. Values are shown as mean ± SEM; * p < 0,05 versus CV; **p < 0,01 versus CV;
n = 3 animals per group.

Fig. 2. Effects of leptin stimulation in mice supplemented, for 8 weeks, with coconut oil (CO100 or CO300) or water (CV). (A) Food intake after 3 h of leptin
stimulation (5 mg/g, IP); (B) Energy expenditure after 3 h of leptin stimulation (5 mg/g, IP); (C) Respiratory Quotient (RER) after 3 h of leptin stimulation (5 mg/g,
IP); Values are shown as mean ± SEM; * p < 0,05 versus respective basal group; **p < 0,01 versus respective basal group; ***p < 0,001 versus respective basal
group; ### p < 0,001 versus CV; ƚ p < 0,05 versus CV + Leptin; n = 4–6 animals per group.

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Fig. 3. Effects of leptin stimulation in the hypothalamus of mice supplemented, for 8 weeks, with coconut oil (CO100 or CO300) or water (CV). (A-C)
Western-blotting analysis of Leptin pathways markers in the hypothalamus of mice supplemented with coconut oil or water after leptin stimulation (2.5 mg/g, IP);
(D-F) mRNA expression of neuropeptides, in the hypothalamus of mice supplemented with coconut oil or water after leptin stimulation (2.5 mg/g, IP), AGRP, NPY
and POMC, respectively. Values are shown as mean ± SEM; * p < 0,05 versus respective basal group; **p < 0,01 versus respective basal group; ***p < 0,001 versus
respective basal group; # p < 0,05 versus CV; ## p < 0,01 versus CV; ### p < 0,001 versus CV; ƚ p < 0,05 versus CV + Leptin; ƚƚ p < 0,01 versus CV + Leptin; ƚƚƚ p <
0,001 versus CV + Leptin; n = 3–6 animals per group.

Additionally, insulin signaling in hypothalamus, white adipose tissue nitrogen and stored at − 80 ◦ C until processing.
and liver was evaluated.
2.3. Leptin and insulin administration
2. Materials and methods
The animals were fasted for 24 h and then stimulated with saline or
2.1. Animals leptin (5 mg/g) to evaluate sensitivity to leptin after eight weeks of CO
supplementation, intraperitoneally, at the beginning of the waking
This investigation was conducted in accordance with the National period. During this same period, the animals were stimulated and re-fed
Institute of Health Guide for the Care and Use of Laboratory Animals and to measure food intake, energy expenditure, and respiratory quotient
the guidelines of the Brazilian Society of Science in Laboratory Animals. three hours after the leptin injection (Garcia-Galiano et al., 2017).
All procedures were approved by the Research Ethics Committee for However, to assess leptin signaling at the end of the experimental
Animal Use of the University of Campinas (Protocol number: 5124–1/ period, saline or leptin (2.5 mg/g) was administered intraperitoneally in
2019). Five-week-old male mice of Swiss lineage, weighing approxi­ mice, which were fasting for 12 h, and 30 min after stimulation (Garcia-
mately 20 g, were provided by the Animal Breeding Center of the Uni­ Galiano et al., 2017) the animals were anesthetized and subsequently
versity of Campinas. The animals were kept in individual cages, with ad decapitated.
libitum water and chow diet (NUVILAB® Cr-1-Nuvital, Brazil), in a room To investigate insulin sensitivity in peripheral tissues, at the end of
with controlled temperature (22–24 ◦ C) and a light/dark cycle (12 h). the experiment for 12 h, and 20 min after the stimulus, the animals were
anesthetized and subsequently decapitated. At the end of the supple­
2.2. Experimental design mentation period the hypothalamus was collected after anesthesia and
decapitation of mice in order to investigate insulin sensitivity in the
The mice (n = 60) were randomly distributed into three groups and central nervous system. Then, hypothalamic tissue was exposed to
received oral supplement with pipette tip, for eight weeks with 300 μL of DMEM with or without insulin (50 nM) for 10 min.
water for the control group (CV, n = 20), 100 or 300 μL of commercial
extra-virgin coconut oil (Copra brand) (CO100, n = 20 and CO300 n = 2.4. Energy expenditure
20, respectively). The supplementation was calculated according to the
recommendation of saturated fat intake, corresponding to a maximum of At the end of the eighth week of supplementation and after leptin
10% of the diet (Reeves et al., 1993), that is, 100 μL. Meanwhile, 300 μL stimulation (5 mg/g), the animals were tested in the Comprehensive Lab
is a volume recommended to mimic the additional consumption of CO. Animal Monitoring System (CLAMS, Columbus Instruments, Columbus,
At the end of the experimental period, after 12 h overnight -fasting, the OH), measuring energy expenditure (expressed in Heat – kcal/min
mice were anesthetized with a mixture containing (Ketamine 100 mg/ /body weight) and respiratory quotient (RER - VO2/O2 ratio) by indi­
kg; Xylazine 5 mg/kg, ip) and then decapitated. Samples of hypothala­ rect calorimetry. CLAMS was installed under constant room temperature
mus, liver and epididymal adipose tissue were weighted, frozen in liquid (22 ◦ C) and light/dark cycle (12 h). For this assay, mice were previously

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Fig. 4. Effects of leptin stimulation in the epididymal adipose tissue of mice supplemented, for 8 weeks, with coconut oil (CO100 or CO300) or water (CV).
(A-E) Western-blotting analysis of Leptin pathways markers in the epididymal adipose tissue of mice supplemented with coconut oil or water after leptin stimulation
(2.5 mg/g, IP); (F) ACC mRNA expression. * p < 0,05 versus respective basal group; **p < 0,01 versus respective basal group; ***p < 0,001 versus respective basal
group; # p < 0,05 versus CV; ## p < 0,01 versus CV; ### p < 0,001 versus CV; ƚ p < 0,05 versus CV + Leptin; ƚƚ p < 0,01 versus CV + Leptin; ƚ ƚ ƚ p < 0,001 versus
CV + Leptin; $$ p < 0,01 versus CO100; && p < 0,01 versus CO100 + Leptin; n = 3–6 animals per group.

adapted for 12 h in individual cages. materials were removed by centrifugation (12,000 rpm) for 30 min at
4 ◦ C. The protein concentration of the supernatant was determined using
2.5. qPCR- real time analysis the Biuret dye-binding method for adipose tissue and liver, and Bradford
assay (Bradford, 1976) for the hypothalamus. The supernatant was
Frozen epididymal adipose tissue (100 mg) and hypothalamus were suspended in Laemmli sample buffer and boiled for 5 min before sepa­
homogenized using Trizol reagent (Invitrogen Corporation, CA, USA). ration by SDS-PAGE using a miniature slab gel apparatus (BioRad,
Total RNA was extracted according to the manufacturer’s guidelines and Richmond, CA, USA). Electrotransfer of proteins from the gel to a
quantified on a Nanodrop ND-2000 spectrophotometer (Thermo Elec­ nitrocellulose membrane was performed for 120 min at 120 V (constant)
tron, WI, USA). Reverse transcription was performed with 3 μg of total in a transfer buffer with methanol. The membranes were subsequently
RNA and a High-Capacity cDNA Reverse Transcription kit (Life Tech­ blocked with a 5% skim milk solution in Tris-buffered saline (TBS)-
nologies Corporation, Carlsbad, CA, USA). Relative expression was Tween 20 (TTBS; 10 mmol Tris/L,150 mmol NaCl/L, 0.5% Tween 20)
determined using the TaqMan PCR Master Mix (Applied Biosystems) and for 2 h at room temperature. After blocking, the membranes were
all primers were obtained from the Applied Biosystems: NPY washed three times with TBST for 5 min each and then incubated
(Mm01410146_m1), POMC (Mm00435874_m1), AGRP overnight at 4 ◦ C with specific primary antibodies, from Cell Signaling,
(Mm00475829_g1), ACADvl (Mm0044293_m1), ACADm pSTAT3 (9145S), pJAK2 (3776S), pAMPK (2535S), pAKT (4060S),
(Mm01323360_g1), AGPAT (Mm00479699_g1), ACACA pEIF2a (9721), from Imgenex, ATF6 (IMG273), from Abcam, pIRE
(Mm01304277_m1), and beta-actin (4351315) or GAPDH (4351309) as (ab48187) and from Santa Cruz, pACCα (sc-30447-R), ACADvl (sc-
endogenous control. Real-time PCR was performed in an ABI Prism 7700 376239), FAS (sc-20140), GRP78 (sc-13968), pPERK (sc-32577),
sequence detection system (Applied Biosystems). Each PCR reaction GAPDH (T5168) as endogenous control. Then, the membranes were
contained 20 ng of cDNA. Data were analyzed using Sequence Detection washed three times with TBST for 5 min and incubated for 2 h at room
System 2.0.5 (Life Technologies Corporation, Carlsbad, CA, USA) and temperature with secondary antibodies diluted in TTBS containing 3%
expressed as relative values determined by the comparative threshold dry skimmed milk). Proteins were detected by chemiluminescence
cycle (Ct) method (2DDCt) according to the manufacturer’s guidelines. (Super Signal West Pico Chemiluminescent Substrate, ThermoFisher
Scientific, MA, USA). Band intensity was determined by the software
application ImageJ and the results are expressed as the ratio between
2.6. Western blotting analysis
specific protein and endogenous protein (GAPDH).

Frozen epididymal adipose tissue (100 mg), liver (100 mg) and hy­
pothalamus were homogenized in freshly prepared ice-cold buffer [1% 2.7. Statistical analysis
(v/v) Triton X-100, 0.1 M Tris, pH 7.4, 0.1 M sodium pyrophosphate,
0.1 M sodium fluoride, 0.01 M EDTA, 0.01 M sodium vanadate, 0.002 M All data were expressed as means ± SEM. Statistical analysis was
PMSF and 0.01 M aprotinin] using a tissue homogenizer (Bead Ruptor initially conducted by applying the Kolmogorov-Smirnov to certify
12 Homogenizer, Omni International, Kennesaw, GA, USA). Insoluble normality. Then, the following was applied: Student’s t-test for

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Fig. 5. Effects of leptin stimulation related to fatty acid metabolism in the epididymal adipose tissue of mice supplemented, for 8 weeks, with coconut oil
(CO100 or CO300) or water (CV). (A-B) Western-blotting analysis of ACADvl and FAS, respectively. (C-D) mRNA expression of genes related to fatty acid meta­
bolism in the epididymal adipose tissue of mice supplemented with coconut oil or water after leptin stimulation (2.5 mg/g, IP). Values are shown as mean ± SEM; * p
< 0,05 versus respective basal group; **p < 0,01 versus respective basal group; ***p < 0,001 versus respective basal group; # p < 0,05 versus CV; ## p < 0,01 versus
CV; ### p < 0,001 versus CV; ƚ p < 0,05 versus CV + Leptin; ƚƚ p < 0,01 versus CV + Leptin; ƚ ƚ ƚ p < 0,001 versus CV + Leptin; $$ p < 0,01 versus CO100; && p <
0,01 versus CO100 + Leptin; n = 3–6 animals per group.

comparisons between two independent groups and one-way analysis of 3. Results

variance (ANOVA) followed by Tukey’s test for comparison between all
experimental groups. GraphPad Prism 8 was used for all statistical an­ 3.1. Coconut oil consumption for 8 weeks induces reticulum stress in the
alyses. Also, p-values < 0.05 were considered statically significant. hypothalamus

CO supplementation decreased GRP78 protein level (Fig. 1B) and

increased expression of ATF6 (Fig. 1F) in the hypothalamus. Moreover,

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Fig. 6. Effects of insulin stimulation in mice supplemented, for 8 weeks, with coconut oil (CO100 or CO300) or water (CV). (A-D) pAKT expression in the
hypothalamus, liver, and epididymal adipose tissue, respectively. Values are shown as mean ± SEM; **p < 0,01 versus respective basal group; ***p < 0,001 versus
respective basal group; ƚƚ p < 0,01 versus CV + Insulin; n = 3 per group.

Fig. 7. Summary of the effects of coconut oil supplementation with or without leptin or insulin stimulation in healthy Swiss mice.

CO consumption increased phosphorylation of PERK, EIF2α, and IRE CO supplementation for eight weeks increased basal STAT3 phos­
(Fig. 1B-1E, respectively). These results show that the CO consumption phorylation when compared to the CV group (Fig. 3A). As expected, the
for eight weeks triggers endoplasmic reticulum stress in the hypothal­ presence of leptin increased hypothalamic STAT3 and JAK2 phosphor­
amus of healthy Swiss mice. ylation in animals from the CV group (Fig. 3B and 3C). However, in mice
that consumed CO the hypothalamic STAT3 and JAK2 phosphorylation
3.2. Coconut oil supplementation alters leptin function and signaling stimulated by leptin was lower than CV + Leptin mice (Fig. 3B and C).
pathway in the hypothalamus Furthermore, we evaluated the capacity of leptin to modulate neu­
ropeptides expression in the hypothalamus. The CO consumption
Three hours after leptin stimulation, we identified a reduction in increased NPY and AGRP mRNA expression (Fig. 3D and 3E). The leptin
food intake and an increase in energy expenditure in the CV group. reduced NPY and AGRP transcripts in all experimental groups (Fig. 3D
However, leptin could not change the food intake and energy expendi­ and 2E). However, the leptin was only able to increase POMC gene
ture of animals supplemented with CO (Fig. 2A and B). Furthermore, expression in the CV group (Fig. 3F).
after leptin administration, the respiratory coefficient was lower in all
experimental groups (Fig. 2C).

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Fig. A1.

Fig. A2.

3.3. Coconut oil supplementation changes leptin signaling pathway and induced ACC phosphorylation (Fig. 4E and F), similarly to that observed
fatty acid metabolism in the epididymal adipose tissue for AMPK.
Furthermore, we evaluated the proteins related to fatty acid meta­
The supplementation with CO did not modify the basal phosphory­ bolism in the epididymal adipose tissue. In CO100 mice the basal protein
lation of STAT3 and JAK2 (Fig. 4B and C) in the epididymal adipose and transcript level of FAS and AGPAT mRNA were not different from
tissue. JAK2 phosphorylation stimulated by leptin was significantly control mice (CV). However, in CO300 mice FAS and AGPAT were
reduced in CO100 and CO300 compared to CV mice (Fig. 4C). However, increased compared to CV mice (Fig. 5A, B and C). Leptin stimulation
STAT3 phosphorylation stimulated by leptin significantly increased in reduced significantly AGPAT mRNA in all groups evaluated (CV and
the CV, CO100, and CO300 mice, with a smaller increase in the CO300 CO), while FAS expression was increased by leptin stimulation (Fig. 5A,
group, as shown in Fig. 4B. Furthermore, the supplementation with CO B and C).
reduced AMPK phosphorylation, but did not reduce the capacity of Regarding fatty acid oxidation, ACADvl and ACADm expression were
leptin to stimulate AMPK phosphorylation compared to the CV group similarly modulated by CO supplementation. While the ACADvl was
(Fig. 4D). The level of total ACC and pACC, a target protein of AMPK, induced in CO100 mice, ACADm was increased in CO300 mice (Fig. 5D,
was not altered by CO supplementation and the stimulus with leptin E and F). After leptin administration, the protein expression of ACADvl

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Fig. A3.

Fig. A4.

and ACADm increased in both CV and CO100 groups, but not in the phosphorylation was lower than that observed in the CV and CO100
CO300 group (Fig. 5D, E and F). mice (Fig. 6A, B and C).

4. Discussion
3.4. Coconut oil consumption does not induce insulin resistance
In an initial study, we found that healthy Swiss mice supplemented
Basal AKT phosphorylation after supplementation with CO (CO100 with CO for eight weeks showed greater weight gain, increased body fat
and CO300) was similar to CV mice in all tissues investigated (hypo­ percentage and activation of inflammatory pathways, depending on
thalamus, liver, and epididymal adipose tissue). The stimulus with in­ TLR4, at the hypothalamic and peripheral tissues (Veras et al., 2021). In
sulin increased significantly AKT phosphorylation in the hypothalamus, addition, food intake of control and the supplemented groups were not
liver, and epididymal adipose tissue of CO100 compared to CV mice different, and we stress that the caloric gain at the end of the 8 weeks of
(Fig. 6A, B and C). However, in CO300 mice the insulin-induced

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Fig. A5.

supplementation was only 50.4 and 151.2 kcal for the animals that inflammatory and ERS pathways in mice, probably through activation of
consumed 100 uL and 300 uL of oil, respectively. Based on these pre­ TLR4 by SFAs, as demonstrated in previous study (Veras et al., 2021).
vious findings and knowing that dietary fat consumption can trigger One of the consequences of ERS activation is the detrimental effect on
endoplasmic reticulum stress (ERS) (Milanski et al., 2009) we initially the ability of leptin and insulin to modulate glucose and energy
aimed to investigate whether CO supplementation for eight weeks homeostasis.
would activate endoplasmic reticulum and oxidative stress in the hy­ One of the mechanisms that triggers leptin resistance is hypotha­
pothalamus of healthy mice. Thus, we have shown that consumption of lamic inflammation (Cui et al., 2017; Velloso and Schwartz, 2011) and
CO induces hypothalamic ERS activation, by increasing protein ERS emerged as a key factor in obesity-associated inflammation and
expression of ATF6 and phosphorylation of IRE, PERK and EIF2α, but leptin resistance (Cui et al., 2017). ERS is activated in several tissues of
not oxidative stress, since there was no difference in the levels of MDA in obese and leptin-resistant animals, including adipose tissue and the
the liver and NFR2 transcript levels in the hypothalamus of mice sup­ brain (Ozcan et al., 2004; Zhang et al., 2008; Ozcan et al., 2009). Hence,
plemented with CO (data not show). It is known that consumption diets we evaluated sensitivity to leptin after eight weeks of CO supplemen­
high in animal fat (SFA rich) induces hypothalamic inflammation and tation and observed that leptin could not change food intake or increase
subsequently ERS (De Souza et al., 2005; Carraro et al., 2018). However, the energy expenditure of animals that consumed coconut oil, suggest­
here we showed that the consumption of coconut oil also activates ing hypothalamic leptin resistance. This hypothesis was confirmed by

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Fig. A6.

Fig. A7.

observing impairment in the leptin signaling, including the reduced 2005), resulting in ERS in the hypothalamus, which, in turn, can cause
effect on POMC expression, and phosphorylation of STAT3 and JAK2 in leptin resistance(Zhang et al., 2008).
the hypothalamus and adipose tissue of mice that consumed coconut oil. Based on the previous outcome and the fact that leptin induces
Previous studies showed that pharmacological inhibition of ERS in vitro lipolysis (Stern et al., 2016), fatty acid oxidation (William et al., 2002),
improves leptin signaling (Hosoi et al., 2008; Ozcan et al., 2008), as well and decreases lipogenesis by ACC-1 phosphorylation (Huang et al.,
as the inhibition of ERS either by genetic or pharmacological improves 2006), we investigated whether the administration of exogenous leptin
leptin sensitivity and decreases both food intake and body weight would be able to promote changes in fatty acid metabolism in the adi­
(Zhang et al., 2008; Ozcan et al., 2008). Recently, we showed that mice pose tissue of mice that consumed CO. Leptin increased the expression
consuming coconut oil presented increased expression of pro- and the transcription of enzymes associated with fatty acid oxidation
inflammatory cytokines (IL-6 and TNF-α) as well as activation of (ACADvl and ACADm) in the CO100 group alone. On the other hand,
TLR4-dependent inflammatory pathways in the hypothalamus and adi­ leptin stimulation, associated with coconut oil supplementation, led to
pose tissue (Veras et al., 2021). Thus, we believe that the leptin resis­ an increase in ACC phosphorylation, probably inhibiting fatty acid
tance triggered by coconut oil consumption is induced by inflammatory synthesis. Although coconut oil consumption reduced AMPK phos­
cytokines and the activation of ERS, given that the consumption of a phorylation favoring lipogenesis, leptin was able to reverse this
high-fat diet (HFD) increases the expression of pro-inflammatory cyto­ parameter. Eventually, AMPK is the major mediator of leptin effects on
kines in the hypothalamus, such as IL-1, IL-6 and TNF-α, activating in­ fatty-acid metabolism (Minokoshi et al., 2002). The predominant effects
flammatory pathways, such as the NFκB pathway (De Souza et al., of leptin on adipose tissue may be mediated by the peripheral nervous

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system (Stern et al., 2016), regardless of the direct action of leptin in the Acknowledgements
adipose tissue. Leptin increases the sympathetic efferent signal to white
adipose tissue to increase lipolysis (Geerling et al., 2014). The authors would like to thank the team at the Laboratory of
Finally, a correlation between insulin and leptin can be established. Metabolic Disorders for their support and helpful advice and Coordi­
Insulin is capable of inducing leptin secretion (Vital et al., 2006; Inui nation for the Improvement of Higher Education Personnel – Brazil
et al., 2012), while leptin activates the main insulin signaling pathway, (CAPES) – Finance Code 001 and the São Paulo Research Foundation
the phosphatidylinositol 3-kinase (PI3K) pathway (Niswender et al., (FAPESP) (grant # 16/23484–1, # 18/01863–6, and #19/07615–7) and
2003), via phosphorylation of insulin receptor substrates by JAK2 Obesity and Comorbidities Research Center (OCRC) of the São Paulo
culminating in PI3K activation. Leptin or LepR deficiency produces a Research Foundation (FAPESP).
phenotype of hyperphagia, obesity and insulin resistance, and leptin
infusion at low doses, not affecting body weight, can correct hypergly­ Appendix A
cemia and hyperinsulinemia in ob/ob mice (Pelleymounter et al., 1995).
Furthermore, increased leptin sensitivity of LepR-expressing cells does See Figs. A1-A7.
not prevent diet-induced obesity, but protects mice from insulin resis­
tance induced by obesity (Pedroso et al., 2014). We observed a scenario Appendix B. Supplementary material
of insulin resistance in mice that consumed coconut oil (CO300), but not
in CO100 mice, which may require a longer exposure to coconut oil to be Supplementary data to this article can be found online at https://doi.
triggered. Since adiposity plays an important role in insulin resistance, it org/10.1016/j.jff.2023.105600.
is difficult to determine whether leptin signaling in a specific brain
structure directly controls glucose homeostasis or whether the observed References
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