Biomedicine & Pharmacotherapy 126 (2020) 110110

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Biomedicine & Pharmacotherapy

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

Investigation of the effects of dietary modification in experimental obesity: T

low dose of virgin coconut oil has a potent therapeutic value
Wale Johnson Adeyemia,*, Luqman Aribidesi Olayakib, Tahir Ahmad Abdussalamc,
Abosede Pelumi Toriolab, Akeem Babatunde Olowub, Adebayo Jamiu Yakubb, Aliu Olayinka Rajib
a
Physiology Department, Redeemer’s University, Ede, Nigeria
b
Physiology Department, University of Ilorin, Ilorin, Nigeria
c
Anatomy and Physiology Department, University of Ilorin Teaching Hospital, Ilorin, Nigeria

A R T I C LE I N FO A B S T R A C T

Keywords: There is no report in literature on possible physiological changes that accompany dietary modification in obese
Virgin coconut oil condition. Moreover, there is no conclusive evidence on the optimal amount of virgin coconut oil (VCO) that
High fat diet could be of health benefit, although it is known to enhance lipid metabolism. Therefore, we investigated the
Lipid profile antiobesitogenic action of graded doses of VCO (200, 400 and 600 mg/kg) in obese rats fed with normo/hyper-
Inflammation
lipidaemic diet. Sixty rats (n = 10) were divided into 6 groups and treated as follows: the control and high fat
Antioxidants
diet (HFD) groups were administered normal saline (0.1 mL/day, p.o.) during the last four weeks of the study,
Liver
and were fed with normal and HFD respectively throughout the twenty weeks duration of the experiment.
Groups 3–6 were fed with HFD for 16 weeks, then normal diet during the next 4 weeks. While group - 3 received
saline (0.1 mL/day, p.o.) during the last four weeks, groups 4–6 received graded doses of VCO. The results
showed that HFD-induced obesity caused impaired glucose homeostasis, distorted hepatic histoarchitecture,
selected deviations in hepatic function indices, pro-inflammatory, pro-oxidant, and dsylipidaemic effects. There
were evidence of escalated and reversed pathological actions following the replacement of HFD with normal
diet. VCO showed no effect on glucose, insulin, insulin resistance, total protein, uric acid and TAC; but equitable
effects on CAT, IL-6, CRP, ALT, AST & GGT, irrespective of the dose. Compared to the effects of VCO at 400 and
600 mg/kg, at 200 mg/kg, VCO had more significant therapeutic effects on LDH, MDA, SOD, GPX, TC, TG, LDL-
C, total bilirubin, atherogenic and lee indices and hepatic histoarchitecture. Conclusively, VCO, preferably at a
low dose could be used to reverse hepatic structural alteration and some biochemical deviations following
dietary modifications in obese condition.

1. Introduction
The global prevalence of obesity and overweight has increased
significantly since 1980 to an extent that about one third of the popu-
lation in the world is now considered to be obese or overweight [1].
Obesity remains a public health threat as it adversely affect virtually all
physiological functions of the body. The disease negatively affects work
productivity, healthcare costs and the general quality of life. Obesity is
a multifactorial disease condition that is secondary in part to long-term
positive energy balance i.e. when energy expenditure is less than
dietary energy intake. The excess energy is converted to triglyceride,
which is stored in the adipose tissue, thereby resulting in increase body
fat and hence weight gain. Specifically, passive overconsumption of
energy-dense, nutrient-poor, processed and affordable foods and
beverages and decrease in physical activity [2] have been identified as a
major driver of obesity epidemic [3]. The disease elevates the risk of
developing ailments such as cancers [4], musculoskeletal disorder [5],
diabetes mellitus [6], cardiovascular disease [7], poor mental health
[8], to mention a few.
Obesity has been associated with oxidative stress, pro-inflammatory
condition, impaired lipid metabolism, glucose dyshomeostasis [9], he-
patic steatosis, compromised hepatic function [10], among others.
Hence, different pharmacological agents have been synthesised to in-
tercept these adverse effects. Some of the FDA-approved medications
for weight management in obese state include orlistat, phentermine
plus topiramate, lorcaserin, bupropion plus naltrexone, liraglutide 3.0,
phentermine alone, and diethylpropion. Each of these medications is
associated with notable side effects. For example, the administration of

⁎
Corresponding author.
E-mail address: adeyemiwalej@gmail.com (W.J. Adeyemi).

https://doi.org/10.1016/j.biopha.2020.110110
Received 10 January 2020; Received in revised form 13 March 2020; Accepted 17 March 2020
0753-3322/ © 2020 Published by Elsevier Masson SAS. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/BY-NC-ND/4.0/).
W.J. Adeyemi, et al. Biomedicine & Pharmacotherapy 126 (2020) 110110

Table 1
Normal and high fat feed formulation.
Normal diet formulation High fat diet formulation Energy value

Ingredient Amount (Kg) Amount expressed in percentage (%) Amount (Kg) Amount expressed in percentage (%) per 100 g (Kcal)

Maize 2.75 17.67 2.75 17.67 0.125

Wheat offal 0.25 1.61 0.25 1.61 0.104
Groundnut cake 2.75 17.67 2.75 17.67 0.444
Fish meal 0.50 3.21 0.50 3.21 0.105
Animal fat (lard) – – –6.25 40.16 0.896
Palm kernel cake 2.5 16.06 2.5 16.06 0.862
Bone meal 0.25 1.61 0.25 1.61 0
Methionine 0.125 0.80 0.125 0.80 0
Lysine 0.125 0.80 0.125 0.80 0
Common salt 0.03125 0.20 0.03125 0.20 0
Vitamin premix 0.03125 0.20 0.03125 0.20 0

orlistat has been reported to cause gastrointestinal side effects such as
fatty/oily stool, increase defecation, feacal urgency, feacal incon-
tinence, oily spotting, among others, which constitute the primary
reason for the discontinuation of the therapy [11]. Therefore, studies
are conducted to investigate the therapeutic value of other promising
readily available substances in the management of obesity.
Initially, it was believed that coconut fats are bad for health, espe-
cially because about 92 % of its fats are saturated. However, since most
of the saturated fats present in coconut oil are medium chain fatty
acids, whose characteristics and metabolism vary from long chain tri-
glycerides (LCT), it is now believed that it might not be as bad as in-
itially thought, when compared to other saturated fats [12]. VCO is a
cheap, readily available, and widely used over-the-counter com-
plementary medicine in many countries of the world [13]. Previous
reports show that VCO has polyphenols [14], which have strong anti-
oxidant property. Moreover, it has been documented that the oil has
therapeutic value in diabetic state [15] and could reduce body fat [16].
Nevertheless, due to the saturated fat content of VCO, there are mixed
report on its effect on lipid metabolism [17,18], which is a core index in
obese state.
There is no report in literature on possible biochemical and struc-
tural changes that accompanies dietary modification in obese condition.
Moreover, the effects of VCO at varying dose in experimental model of
obesity using normo/hyper-lipidaemic diet have not been perused.
Therefore, the aim of this study was to investigate the dose-graded ef-
fects of VCO and also the effects of dietary change from high fat diet to
normal diet in obese male Wistar rats. A wide range of parameters such
as glucoregulatory, antioxidant, and inflammatory markers, Lee and
atherogenic indices, markers of hepatic function, lipid profile and he-
patic histoarchitecture were considered in the present study.
2. Materials and methods
2.2. Animal care
Sixty (60) adult male Wistar rats weighing 150–220 g were pur-
chased from the Animal House of the College of Health Sciences,
University of Ibadan, Ibadan, Nigeria. They were kept in plastic cages at
a room temperature of about 27–30 °C and photoperiodicity of 12 h
light – 12 h dark. The rats were allowed to acclimatise in their new
environment for 2 weeks before the commencement of the experiment
and were given feed and water ad libitum. The experimental procedures
were in accordance with the ethical guidelines of the University of
Ilorin (ethical number was not assigned for this study) and the National
Academy of Sciences Guide for the Care and Use of Laboratory animals
[19].
2.3. Feed formulation
Normal and high fat rat chow were manufactured by Ace Feed PLC
Ibadan, Oyo, Nigeria. The feed formulation and the percentage of each
component are described in the table below (see Table 1).
2.4. Preparation of VCO
Dry coconuts were obtained from the Kwara State Ministry of
Agriculture and Natural Resources, Ilorin, Nigeria. The nuts were
broken and the solid endosperm component of mature coconuts (Cocos
nucifera) was grinded and made into viscous slurry. About 400 mL of
water was added to the latter and then the mixture was squeezed
through a fine sieve so as to obtain coconut milk, which was left for
almost one day to enhance gravitational separation of the emulsion.
Afterwards, the supernatant oil layer was scooped off and the product
was filtered through a fine and sterile sieve and then stored in a clean
bottle at room temperature. The table below shows the phytochemical
constituents of VCO.

2.1. Drugs and chemicals

2.5. Experimental design
Sodium pentobarbital was purchased from Actiza Pharmaceutical
The rats that were used for this study were divided into six (6)
Private Ltd., Surat, Gujarat, India, while assay kits for the determina-
groups of ten rats each (sample size was determined by G* power
tion of lactate dehydrogenase, malondialdehyde, total cholesterol, tri-
software), which included: the control (untreated) group; high fat diet
glyceride, low and high density lipoprotein cholesterol, superoxide
group (HFD); high fat diet recovery group (hfd); high fat diet + 200
dismutase, catalase, glutathione peroxidase, total antioxidant capacity,
mg/kg virgin coconut oil (hfd + VCO 200); high fat diet + 400 mg/kg
interleukin – 6, c-reactive protein and uric acid were purchased from
virgin coconut oil (hfd + VCO 400); and high fat diet + 600 mg/kg
Cayman Chemical company, Michigan, USA. Moreover, analytical kits
virgin coconut oil (hfd + VCO 600). The control and HFD groups were
for the determination of insulin, albumin, total protein, total bilirubin,
administered normal saline (0.1 mL/day, p.o.) during the last four
aspartate aminotransferase, alanine aminotransferase, alkaline phos-
weeks of the study, and were fed with normal rat chow and high fat diet
phatase and gamma-glutamyl transferase were procured from Abcam
respectively throughout the twenty weeks duration of the experiment.
PLC, Cambridge, UK. The analyses were performed according to the
Groups 3–6 were fed with HFD for 16 weeks, then normal diet for the
manufacturers’ instruction.
next 4 weeks. While, group 3 received normal saline (0.1 mL/day, p.o.)
during the last four weeks, groups 4–6 received VCO at 200, 400 and

2
W.J. Adeyemi, et al.
600 mg/kg B.W respectively.
Obesity was confirmed in the rats by using Lee index. It was cal-
culated by using the formula: 4√body weight (g) / nose-anal length
(cm). At the end of the 16 weeks of administration of high fat diet,
animals with an index equal to or lower than 0.30 were considered non-
obese and were not used for the study, while rats with an index higher
than 0.30 were considered to be obese [20] and were selected and
randomized into the different study groups.
About twenty-four hours after the last treatment at the end of the
twenty weeks duration of the experiment, all the animals were sacri-
ficed under sodium pentobarbital anaesthesia (10 mg kg−1 body
weight, i.m.) [21–23]. Blood samples were collected by cardiac punc-
ture into lithium heparinised tubes, which were centrifuged at 3500
rpm for 15 min at −4 °C using a cold centrifuge (Centurion Scientific
Ltd., West Sussex, UK, Model 8881). The separated plasma samples
were collected into separate plain tubes for the assessment of bio-
chemical markers. Moreover, the hepatic tissues were excised, washed
in ice cold phosphate buffered saline and subsequently fixed in a buffer
solution of 10 % formalin prior to the histological analysis.
2.6. Determination of blood glucose and insulin resistance
Terminal blood glucose level was determined using Accu – Check
Active (manufacturer: Roche Diagnostics, Pvt Ltd., Mumbai,
Maharashtra, India), while insulin resistance was determined by using
the formula:
Fasting blood glucose (mmol/l) × fasting serum insulin (pmol/l) /
22.5 as described by [24,25].
2.7. Histopathological assessment of hepatic tissue
As described in our previous study [26], the fixed hepatic tissues in
10 % formalin were dehydrated in graded concentration of ethanol,
cleared in xylene and embedded in paraffin wax. The tissues were cut
into 4 μm thick sections using a microtome and then fixed on slides,
prior to staining with haematoxylin–eosin. The stained slides were ex-
amined under an optical microscope (Olympus CH; Olympus, Tokyo,
Japan) and the photomicrographs were obtained with a Leica DM 750
camera at ×40 magnification.
2.8. Statistical analysis
Data obtained from the study were expressed as mean ± standard
error of mean (SEM). Analyses were performed by one way analysis of
variance (ANOVA), followed by Tukey post-hoc test, using IBM SPSS
software, version 20.0. Values were considered to be statistically sig-
nificant at p < 0.05.
3. Results
3.1. Effects of graded doses of VCO on lactate dehydrogenase (LDH),
malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT),
glutathione peroxidase (GPX), total antioxidant capacity (TAC), interleukin
– 6 (IL-6), c-reactive protein (CRP) and uric acid (UA) in obese Wistar rats
Compared to the control group, significant increases were recorded
in LDH activities (Fig. 1a) in hfd rec and hfd + VCO 400 (p – 0.002 and
0.001 respectively); however, significant decreases were observed in
MDA level (Fig. 1b) in hfd rec, hfd + VCO 200 and hfd + VCO 600
groups (p - 0.009, 0.000 and 0.037 respectively), in SOD activities
(Fig. 1c) in HFD, hfd rec, hfd + VCO 400 and hfd + VCO 600 (p –
0.000), and in CAT activities (Fig. 1d) in HFD, hfd + VCO 200, hfd +
VCO 400 and hfd + VCO 600 groups (p – 0.001, 0.016, 0.001 and 0.000
respectively). Moreover, relative to the control group, there were sig-
nificant reductions in GPX activities (Fig. 1e) in hfd + VCO 400 and hfd
+ VCO 600 (p – 0.012 and 0.019 respectively) and in TAC (Fig. 1f) in
3
Biomedicine & Pharmacotherapy 126 (2020) 110110
groups 2–6 (p – 0.000). There was a significant reduction in MDA level
in hfd rec, relative to HFD group (p – 0.006). In addition, in comparison
to group 3 (hfd), there were significant decreases in MDA level in hfd
+VCO 200 (p – 0.002), and in LDH activity in hfd +VCO 200 (p –
0.014) and also in CAT activity in hfd +VCO 600 (p - 0.014); never-
theless, a significant increase was recorded in SOD activity in hfd +
VCO 200 group (p – 0.000). Relative to the control group, there were
significant elevations in IL-6 (Fig. 1g) and CRP (Fig. 1h) in hfd rec (p –
0.017 and 0.049 respectively), and in UA (Fig. 1i) in HFD group (p –
0.000); however, a significant decrease was noted in the UA level in hfd
+ VCO 600 group (p – 0.017). In comparison to HFD group, there was a
significant elevation in IL-6 level in hfd rec group (p – 0.005); but, a
significant decrease was documented in UA in the hfd rec group (p –
0.000). Moreover, relative to hfd rec group, there were significant di-
minution in the level IL-6 and CRP in hfd +VCO 200, hfd +VCO 400
and hfd +VCO 600 groups (p – 0.000, 0.000, 0.000; 0.000, 0.005 and
0.016 respectively).
3.2. Effects of graded doses of VCO on blood glucose, insulin, insulin
resistance, total cholesterol (TC), triglyceride (TG), and low density
lipoprotein cholesterol (LDL-C) in obese Wistar rats
There were significant elevation in the glucose, insulin and insulin
resistance (Table 2) in HFD group compared to the control group (p –
0.000, 0.032 and 0.001 respectively). However, relative to the latter,
significant decreases were observed in the insulin resistance in hfd
+VCO 200, hfd +VCO 400 and hfd +VCO 600 (p – 0.049, 0.001 and
0.043 respectively). In comparison to the HFD group, there were sig-
nificant decreases in glucose, insulin and insulin resistance in hfd rec
group (p – 0.000, 0.033, and 0.000 respectively) (Table 2). There was
no significant difference in the HDL-C level when comparisons were
made among the groups (Table 2). However, compared to the control
group, there were significant elevations in TC and LDL-c (Table 2) in
HFD, hfd rec, hfd + VCO 400 and hfd + VCO 600 (p – 0.000, 0.000,
0.000 and 0.000; 0.000, 0.000, 0.002 and 0.002 respectively), and in
TG (Table 2) in hfd rec and hfd + VCO 600 groups (p – 0.006 and 0.004
respectively). In addition, relative to the control group, there were
significant increases in atherogenic index (Table 3) in hfd rec, hfd +
VCO 400 and hfd + VCO 600 (p – 0.000, 0.005 and 0.022 respectively)
and in Lee index (Table 3) in HFD, hfd rec, hfd + VCO 400 and hfd +
VCO 600 (p – 0.045, 0.043, 0.042 and 0.044 respectively). Relative to
HFD group, there was a significant elevation in atherogenic index in hfd
rec group (p – 0.001). It is noteworthy to likewise state that, compared
to the latter, significant decreases were recorded in TC, TG, LDL-C and
atherogenic index in hfd +VCO 200 group (p – 0.000, 0.026, 0.000 and
0.000 respectively).
3.3. Effects of graded doses of VCO on high density lipoprotein cholesterol
(HDL-C), atherogenic index, lee index, albumin, total protein, total bilirubin
(TB), aspartate aminotransferase (AST), alanine aminotransferase (ALT),
alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) in
obese Wistar rats
Just like the case of HDL-C, no significant difference was noted in
the albumin level when comparisons were done across the groups
(Table 4). Nevertheless, relative to the control group, there were sig-
nificant diminutions in total protein (Table 4) in groups 2–6 (0.048,
0.021, 0.018, 0.008 and 0.008 respectively), and in AST and ALT
(Table 4) in groups 4–6 (p – 0.018, 0.039 and 0.021; 0.000, 0.000 and
0.000 respectively). Nevertheless, in comparison to the control group,
there were significant increases in the activities of AST and ALP
(Table 4) in HFD and hfd rec (p – 0.002 and 0.000; 0.000 and 0.045
respectively), and in TB (Table 4) in HFD group (p – 0.001) and also in
GGT (Table 4) in hfd rec group (p – 0.001). Moreover, relative to the
HFD group, a significant decrease was recorded in ALP activity in hfd
rec group (p – 0.014); however, a significant elevation was documented
W.J. Adeyemi, et al. Biomedicine & Pharmacotherapy 126 (2020) 110110

Fig. 1. Effects of graded doses of VCO on lactate dehydrogenase (LDH) (Fig. 1a), malondialdehyde (MDA) (Fig. 1b), superoxide dismutase (SOD) (Fig. 1c), catalase
(CAT) (Fig. 1d), glutathione peroxidase (GPX) (Fig. 1e), total antioxidant capacity (TAC) (Fig. 1f), interleukin – 6 (IL-6) (Fig. 1g), c-reactive protein (CRP) (Fig. 1h)
and uric acid (UA) (Fig. 1i) in obese Wistar rats.
Values are expressed as mean ± SEM. *p≤ 0.05 is significant compared to control group; #p ≤ 0.05 is significant compared to HFD group; αp≤ 0.05 is significant
compared to hfd rec group.

in the activity of GGT in the same group (p – 0.005). Compared to hfd
rec group, significant decreases in AST and GGT were observed in hfd
+VCO 200, hfd +VCO 400 and hfd +VCO 600 groups (p – 0.000,
0.000 and 0.000; 0.000, 0.003 and 0.002 respectively). Moreover, there
were significant reductions in TB in hfd +VCO 200 (p – 0.005) and also
in ALP in hfd +VCO 600 (p – 0.049) when compared to hfd rec group.
3.4. Effects of graded doses of VCO on hepatic histoarchitecture in obese
Wistar rats
Fig. 2a shows normal central vein (yellow arrows), normal hepa-
tocytes (green arrows), kuffper cells (purple arrows), and overall
normal morphology of the tissue. In the Fig. 2b, the hepatic tissue
shows steatosis (blue arrows) and ischaemic appearance, shrinked and
compromised integrity of the central vein (yellow arrows), dead he-
patocytes (black arrows), kuffper cells (purple arrows), and foci of in-
flammatory cells (red rings). The Fig. 2c shows no steatotic but
ischaemic appearance of the hepatic tissue, foci of inflammatory cells
(red rings), disappearance of the central vein (yellow arrows), normal
hepatocytes (green arrows), and kuffper cells (purple arrows), while
Fig. 2d shows few but scattered steatotic appearances in the hepatic
tissue (blue arrows), normal hepatocytes (green arrows), kuffper cells
(purple arrows), normal central vein and evidence of well-vascularised
tissue (no ischaemic appearance). Fig. 2e shows wide-spread evidence
of steatosis, especially around the hepatocytes, overall ischaemic ap-
pearance, vasoconstricted and distorted morphology of the central vein
(yellow arrow), normal (green arrows) and dead (black arrows) hepa-
tocytes, and kuffper cells (purple arrows), while Fig. 2f shows no

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W.J. Adeyemi, et al. Biomedicine & Pharmacotherapy 126 (2020) 110110

Table 2
Effects of graded doses of VCO on blood glucose, insulin, insulin resistance, total cholesterol (TC), triglyceride (TG), and low density lipoprotein cholesterol (LDL-C)
in obese Wistar rats.
Groups/Parameters Glucose (mmol/l) Insulin (Pmol/l) Insulin resistance (HOMA-IR) TC (mg/dl) TG (mg/dl) LDL-C (mg/dl)

1. Control 2.576 ± 0.113 0.087 ± 0.004 0.013 ± 0.001 54.246 ± 1.874 48.839 ± 2.0425 58.923 ± 5.277
2. HFD 3.647 ± 0.140* 0.147 ± 0.029* 0.019 ± 0.001* 133.990 ± 2.663* 71.932 ± 8.4126 119.150 ± 4.053*
3. hfd rec 2.576 ± 0.087# 0.087 ± 0.009# 0.010 ± 0.001# 121.990 ± 7.956* 97.246 ± 8.5681* 135.080 ± 8.677*
4. hfd + VCO 200 2.907 ± 0.029 0.072 ± 0.002 0.010 ± 0.000* 58.418 ± 1.365α 57.051 ± 10.0062α 44.454 ± 5.541α
5. hfd + VCO 400 2.423 ± 0.049 0.067 ± 0.002 0.007 ± 0.00* 121.010 ± 4.051* 71.875 ± 10.5723 95.006 ± 3.470*
6. hfd + VCO 600 2.958 ± 0.110 0.071 ± 0.003 0.009 ± 0.001* 120.730 ± 3.520* 99.578 ± 5.9087* 95.496 ± 4.009*

Values are expressed as mean ± SEM.

* p ≤ 0.05 is significant compared to control group.
#
p ≤ 0.05 is significant compared to HFD group.
α
p ≤ 0.05 is significant compared to hfd rec group.

significant evidence of steatosis, tissue degeneration (white arrows),
accumulation of fat in the central vein (yellow arrow), normal (green
arrows) and dead (black arrows) hepatocytes, and kuffper cells (purple
arrows).
4. Discussion
Recently, Dias et al. [27] evaluated the effects of partial substitution
of lipid sources in normolipid and isocaloric diet with varying con-
centrations of VCO in non-obese Wistar rats, with focus on anthropo-
metric index, the composition of the intestinal microbiota, and wide
range of other biomarkers. However, in the present study, we in-
vestigated the dose-graded effects of VCO in obese male Wistar rats. At
large, we observed that VCO has a therapeutic value in obese condition.
Nevertheless, these effects are dose-sensitive, the lowest dose being the
most effective. At higher doses, VCO either showed no effects or ad-
verse actions. Moreover, it causes distorted histoarchitecture of the
hepatic tissue.
As expected, obesity, confirmed by the calculation of Lee index in
the present study was accompanied with dsylipidaemia. Although,
there was no significant difference in the HDL-C across the groups, the
significant increases in TC, TG, LDL-C from the base-line indicated that
there was an imbalance in lipid metabolism. HFD causes an increase in
the accumulation of lipid molecules in the adipose tissue, due to their
high energy content, consequently resulting in an increase in body
weight. It was surprisingly that although obesity was established in this
study, there was delayed significant alterations in TG and atherogenic
index until dietary change. Reports in literature established that VCO
has antidyslipidaemic effect. This has been attributed to its high content
of medium-chain triglycerides (MCT), which is known to enhance lipid
metabolism and improve fatty acid oxidation [28], as a result con-
tributing to reductions in body weight and BMI [29]. In the present
study, it was observed that this effect is dose-related. At 200 mg/kg,
VCO significantly reduced the Lee index to a level comparable to the
control group, and at the same time, it significantly reduced TC, TG,
LDL-C and atherogenic index below values recorded after dietary
change from hyperlipidaemic to normolipidaemic diet. Nevertheless, at
400 and 600 mg/kg, VCO seemed to have no effects on lipid metabo-
lism. Therefore, we suggested that the administered dose of VCO could
be responsible for the lack of relationship between coconut oil and lipid
profile as documented in some studies [30,31].
In obese state, increase in the level of free fatty acid in the plasma
and hence elevated utilisation of lipids by tissues, results in an im-
pairment in glucose metabolism [32]. This will no doubt precipitate
insulin resistance and also dyslipidaemia. Dyslipidaemia is often seen in
association with diabetes and insulin resistance [33–36]. The observed
insulin resistance in the HFD group was not surprising, as hypergly-
caemia and hyperinsulinaemia recorded in the same group are con-
tributory factors. However, dietary change from high fat diet to normal
diet was accompanied with significant reductions in these indices.
Therefore, some detrimental effects that accompany obesity could
simply be overcome by dietary change, without the usage of any
therapeutic agents. At first, it was surprising that graded doses of VCO
showed no effect on glucose, insulin and insulin resistance. But, on the
second look, the expected effects of VCO on glucoregulatory indices was
blunted by dietary change from hyperlipidaemic to normolipidaemic
diet, which was observed to restore glucose homeostasis and hence
promote insulin sensitisation in obese condition, without pharmacolo-
gical intervention, as shown by the comparison of the glucose, insulin,
and insulin resistance results in the HFD group relative to hfd rec group.
Studies have shown that the consumption of VCO [27,37] and
MCFA [38] increases the endogenous level of pro-inflammatory cyto-
kines. Contrarily, in the present study, it was observed that VCO has
anti-inflammatory effect. Unlike what was recorded in the indices of
lipid and glucose metabolism, the anti-inflammatory effects of VCO is
largely not dose sensitive. At the administered doses, VCO significantly
reduced IL-6 and CRP, although it showed no significant effect on the
plasma level of UA. We could not justify why there was a delayed
elevation in CRP and IL-6, but not UA, until there was dietary change.
Obesity is considered as a chronic inflammatory state. Preadipocytes
and adipocytes have been identified as a source of pro-inflammatory
cytokines, such as IL-1, IL-6, TNF-α, among others [39,40]. Therefore,

Table 3
Effects of graded doses of VCO on high density lipoprotein cholesterol (HDL-C), atherogenic index, lee index, albumin and total protein in obese Wistar rats.
Groups/Parameters HDL-C (mg/dl) Atherogenic index log (TG/HDL-C) Lee index [4√BW (g) X nose-anal length (cm)] Albumin (g/dl) Total Protein (g/dl)

1. Control 32.180 ± 4.1677 0.269 ± 0.006 0.27 ± 0.01 6.2147 ± 0.069 12.231 ± 1.150
2. HFD 22.041 ± .9638 0.347 ± 0.001 0.37 ± 0.01* 6.3209 ± 0.148 10.003 ± 0.325*
3. hfd rec 30.306 ± 2.1224 0.580 ± 0.018*,# 0.36 ± 0.00* 6.1274 ± 0.056 9.674 ± 0.168*
4. hfd + VCO 200 31.124 ± 2.2926 0.227 ± 0.037α 0.34 ± 0.01 6.1558 ± 0.115 9.623 ± 0.074*
5. hfd + VCO 400 27.649 ± 2.5401 0.472 ± 0.061* 0.36 ± 0.03* 6.1957 ± 0.070 9.348 ± 0.069*
6. hfd + VCO 600 27.674 ± 2.1818 0.439 ± 0.034* 0.37 ± 0.02* 6.0723 ± 0.053 9.332 ± 0.206*

Values are expressed as mean ± SEM.

* p ≤ 0.05 is significant compared to control group.
#
p ≤ 0.05 is significant compared to HFD group.
α
p ≤ 0.05 is significant compared to hfd rec group.

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W.J. Adeyemi, et al. Biomedicine & Pharmacotherapy 126 (2020) 110110

Table 4
Effects of graded doses of VCO on total bilirubin (TB), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-
glutamyl transferase (GGT) in obese Wistar rats.
Groups/Parameters TB (umol/l) AST (U/L) ALT (U/L) ALP (U/L) GGT (U/L)

1. Control 12.015 ± 0.827 85.205 ± 14.254 20.328 ± 2.233 60.939 ± 10.146 6.254 ± 1.392
2. HFD 18.443 ± 0.875* 142.090 ± 3.032* 20.521 ± 1.683 143.570 ± 12.579* 8.290 ± 1.735
3. hfd rec 15.384 ± 1.280 150.740 ± 13.055* 14.360 ± 1.844 98.843 ± 8.686*,# 16.433 ± 1.819*,#
4. hfd + VCO 200 9.887 ± 0.365α 41.422 ± 1.293*,α 8.574 ± 1.320* 90.864 ± 5.267 4.436 ± 0.851α
5. hfd + VCO 400 13.832 ± 1.256 45.963 ± 5.227*,α 8.013 ± 0.331* 78.508 ± 3.452 7.781 ± 0.899α
6. hfd + VCO 600 12.909 ± 0.334 42.330 ± 3.200*,α 8.382 ± 0.962* 61.521 ± 6.087α 7.272 ± 1.095α

Values are expressed as mean ± SEM.

* ≤p 0.05 is significant compared to control group.
#
≤p 0.05 is significant compared to HFD group.
α
≤p 0.05 is significant compared to hfd rec group.

VCO - enhanced increase in lipid metabolism and the resulting decrease
in adipose tissue mass could explain the observed anti-inflammatory
effects of VCO recorded in this study. At 400 and 600 mg/kg, VCO
showed no significant effect on lipid profile, Lee and atherogenic in-
dices; however, at these doses, it has anti-inflammatory effect. Hence,
the anti-inflammatory effect of VCO is not constrained to the reduction
in preadipocytes and adipocyte tissue mass.
Inflammation and oxidative stress are closely related events.
Increase in the plasma level of inflammatory markers is often accom-
panied with imbalance in antioxidant/pro-oxidant balance in the body
and consequently oxidative stress [41]. The pathways that trigger the
production of inflammatory mediators are all mediated by oxidative
stress. The accumulation of fat in obese state has been linked with
elevation in oxidative stress [42,43]. Therefore, it is not surprising that
obesity is associated with both pro-inflammatory and pro-oxidant
events. The insignificant effect of graded doses of VCO on TAC may
suggests that the oil has no effect on inflammatory status; however, it
could be safely asserted the anti-inflammatory action of VCO is specific
to certain inflammatory markers, as it was observed in the present study
that VCO significantly reduced the plasma levels of IL-6 and CRP. This
effect was not explicit in the UA result in the groups 4–6. Although, at
varying doses VCO showed no effect on TAC, we noted that at 200 mg/
kg, the oil has a significant anti-peroxidative effect – marked by the
determination of MDA. This effect was complemented by significant
reduction in tissue damage and death, evident by the reduction in the
activities of LDH. A positive association was observed in our previous
study between the plasma level and activity of MDA and LDH respec-
tively [44]. Asides, at 200 mg/kg, VCO significantly increased SOD
activity; however, the insignificant effect of the oil on CAT activity and
TAC at the same dose indicates that the VCO has selective actions on

Fig. 2. Effects of graded doses of VCO on hepatic histoarchitecture in obese Wistar rats (H and E X40).
NB: Yellow arrows = central vein; green arrows = normal hepatocytes; black arrows = dead hepatocytes; purple arrows = kuffper cells; blue arrows = steatosis;
white arrows = tissue degeneration; and red rings = foci of inflammatory cells.

6
W.J. Adeyemi, et al.
antioxidant species. The antioxidant effects of VCO could be ascribed to
its terpenoid and anthraquinone constituents, which are known to have
potent antioxidant property [45,46]. Significant reductions in SOD and
GPX activities in the groups that were administered VCO at 400 and
600 mg/kg, and the significant decrease in CAT activity in group 6 (hfd
+ VCO 600) attest to the detrimental effects of VCO at higher doses.
These observed effects were largely supported by the histological as-
sessment of the hepatic tissue. The consequences of dietary change from
high fat diet to normal diet as observed in group 3 (hfd rec) could be
both detrimental and beneficial. Specifically, there were significant
elevations in the level and activity of MDA and LDH respectively, in hfd
rec group relative to HFD group; moreover, there was restoration in
CAT activity in the former to a level comparable to the control group.
As observed in the present study, a positive association was recorded
between MDA level and LDH activity. This is in agreement with our
previous investigations [25,47].
Lim and colleagues [48] indicated that swollen hepatocytes, diffuse
microvesicular and macrovesicular steatosis, and hepatocyte necrosis
characterise HFD - induced obesity in rats. This is in consonance with
the findings of the present study. As expected, there was conspicuous
evidence of hepatic steatosis in the group 2 (HFD), which was accom-
panied with compromised integrity of the central vein, ischaemic ap-
pearance of the tissue, and dead hepatocytes. The accumulated fat in
the liver tissue of the obese rats was soon metabolised after reversion to
normal diet. However, unexpectedly, this was accompanied with the
disappearance of the central vein. This suggests that sudden dietary
change from hyperlipidaemic diet to normolipidaemic diet could
trigger some pathological events. This histological finding was sup-
ported by the IL-6, CRP, atherogenic index, MDA, and LDH results.
There is no research work on the effect of sudden dietary change on
physiological processes; hence we could not relate this finding with
another study. The foci of inflammatory cells in the hepatic tissue of the
hfd rec group complement the significant increase in the level of in-
flammatory markers in the same group. There are few but scattered fat
accumulation in the hepatic tissue of the group treated with 200 mg/kg
of VCO - an evidence of improved lipid metabolism, which is in tandem
with the lipid profile result. Moreover, there was an obvious restoration
of the integrity of the central vein and the vascularised and cellular
appearance of the hepatic tissue in this group. The increase in blood
flow to the hepatic tissue also explains the improved lipid metabolism
in the rats treated with 200 mg/kg VCO. Hence, this study proves that
the impairment in lipid metabolism that characterises obese state is
linked with the compromised status of the hepatic blood vessels.
However, at higher doses of VCO, there was reappearance of steatosis
and ischaemia, and distorted morphology of the central vein. The
presence of steatosis in groups 4 and 5 is in agreement with the lipid
profile result, and the atherogenic and Lee indices. It is noteworthy to
indicate that the histoarchitecture of the hepatic tissue appeared to be
more distorted in the group that was administered 600 mg/kg of VCO.
In this group, there was obvious tissue degeneration, dead hepatocytes
and fat accumulation in the central vein. This result was supported
basically by the TG and CAT results, but only slightly by the SOD ac-
tivity and the TAC. It could therefore be opined that at higher doses,
VCO disturbs the delicate endogenous antioxidant/pro-oxidant balance,
precipitates lipid peroxidation and ultimately causescell death and
tissue degeneration.
It will not be arbitrary to expect that the histological findings of the
hepatic tissue would have a predictive value in the result of various
markers of hepatic function. But, there appears to be a little connection
between the hepatic histoarchitecture and the plasma level of the
markers of hepatic function. VCO showed more effect on the hepatic
structure than indices of hepatic function. The reason for this could not
be ascertained. Nevertheless, we submit that non-assessment of the
markers of hepatic function in the hepatic tissue is one of the limita-
tions of this study. Further studies could look into this. Increase in the
activities of ALP and GGT in the hfd rec group, relative to HFD group,
7
Biomedicine & Pharmacotherapy 126 (2020) 110110
complemented the worsened hepatic structure in the former, when
compared to the latter. Previous studies have shown a positive asso-
ciation between the levels of activity of hepatic enzymes and obesity
[10,49]. Pro-oxidative event, which is secondary to dsylipidaemia in
obese state could instigate hepatic damage. This was manifested by
increased activities of various markers of hepatic function [50]. The
effect of graded doses of VCO on albumin level could not be established
because the hyperlipidaemic diet showed a null effect on this para-
meter. Moreover, there was no significant difference in the effects of
graded doses of VCO on the total protein, AST, ALT and GGT activities.
However, at 200 mg/kg, VCO showed a superior effect on the TB, but at
600 mg/kg, VCO seems to have more potent effect on the ALP activity.
This indicates the selected benefit, although largely detrimental effect
of VCO at higher doses in obese rats fed with normolipidaemic diet.
Based on the experimental protocol presented in this manuscript,
further study could assess food consumption rate, feacal fat content,
body weight gain, adipose tissue environment and hormonal changes
e.g. ghrelin, adiponectin, among others
5. Conclusion
VCO, preferably at a low dose could be used to reverse hepatic
structural alteration and some biochemical deviations in obese rats fed
with normolipidaemic diet.
Research funding
This research did not receive any specific grant from funding
agencies in the public, commercial, or non-profit sectors.
Declaration of Competing Interest
Authors state no potential conflict of interest.
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