Effect of high fat ketogenic diet on some cardiovascular and renal parameters in Wistar albino rats

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Int. J. Biol. Chem. Sci. 12(6): 2703-2712, December 2018
ISSN 1997-342X (Online), ISSN 1991-8631 (Print)
© 2018 International Formulae Group. All rights reserved. 5047-IJBCS
DOI: https://dx.doi.org/10.4314/ijbcs.v12i6.19
Original Paper http://ajol.info/index.php/ijbcs http://indexmedicus.afro.who.int
Effect of high fat ketogenic diet on some cardiovascular and renal
parameters in Wistar albino rats
Eiya Bibiana OMOZEE* and Osakue James OSAMUYIMEN
Department of Physiology, School of Medical Sciences University of Benin, Benin City, Nigeria.
*Corresponding author; E-mail: eiyabibiana@gmail.com, Tel: +237 08081953639
ABSTRACT
Ketogenic diet is used in weight control. This study aimed at determining the effect of the consumption
of this high fat, low carbohydrate diet on plasma viscosity, lipid profile, serum and urine creatinine, urea,
electrolytes and urinary pH. Forty Wistar rats grouped into experimental (A) and control groups (B) were
used. Experimental group was fed with 65% fatty diet; control group was fed with normal rat chow for eight
weeks. Fasting samples were collected at the end of the study for analysis. Biochemical analyses were done
using spectrophotometric method, electrolytes were assayed by the ion electrode selective method, plasma
viscosity was analysed using the Reid and Ugwu method and urinalysis strip was used to measure urinary pH.
A significant increase in serum (0.866±0.031) and urinary (5.470±0.424) creatinine urinary sodium
(147.3±23.60), chloride (136.0±23.90), bicarbonate (1570±1065) and pH (7.60±1.63) were observed in the
experimental group when compared with serum values of control group (0.6600±0.060) and urinary
(1.700±0.230) creatinine, sodium (35.60±18.92), chloride (30±17.05), bicarbonate (800±184.4) and pH
(6.8±0.3). A significant decrease in total cholesterol (95.60±2.64) and LDL Cholesterol (43.60±2.16)
concentrations were observed in the experimental group when compared with the control group, total
cholesterol (132.8±10.7) and LDL Cholesterol (69.00±10.80). This study also showed a significant increase in
weight of control groups (initial weight (g) 135.2±5.115, final weight, 151.2±38.3) while there was no
significant increase in weight of the experimental group (Initial weight 170.6±11.95, final weight170.6±11.21).
There was no significant difference in the other parameters when both groups were compared. These findings
have clearly shown that intake of high fat ketogenic diet is not harmful to the heart, however the increase in
serum creatinine in the experimental group could be as a result of the effect of the diet on the kidney or it could
be due to muscle wastage which might be responsible for maintenance of their body weight. Increased urinary
sodium excretion could lead to increased loss of fluid which can also give an impression of weight loss.
© 2018 International Formulae Group. All rights reserved.
Keywords: Ketogenic diet, Cardiovascular, Renal, plasma viscosity, lipid profile, electrolytes.
INTRODUCTION
Obesity is a major risk factor for most
metabolic syndromes. Asa result, one of the
treatment strategies is weight reduction.
Weight loss is usually used as a means of
therapy and it is aimed at improving some of
the metabolic syndromes. In recent time, the
use of ketogenic diet as a means of weight
control has been on the increase. Ketogenic
diet is characterized by a reduction in
carbohydrates (usually less than 50 g/day) a
relative moderate increase in protein and a

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high fat diet (Veech, 2004). This diet is based
on consumption of long chain saturated
triglycerides in a ratio of 3:1 or 4:1 with its
derivation from fat consumption of about
90%, this implies that majority of its calories
is derived from fat (Kristopher et al., 2007).
Ketogenic diets are of two types, the high fat
ketogenic diet and high protein ketogenic diet.
Long term consumption of Ketogenic diet has
been successfully used in the treatment of
refractory paediatric epilepsy, despite its
likely side effect (Freeman et al., 2006).
Researchers have shown that this diet is not
only useful in weight control but also it has
been shown to result in improvement of
metabolic syndromes (Kennedy et al., 2001).
However the fact that a high fatty diet
might have some detrimental effect on lipid
profile, which might affect plasma viscosity of
individuals on this diet and by implication,
impairs cardiovascular functions should be of
great concern.
Blood viscosity is the intrinsic
resistance of blood to flow in blood vessels in
which its major determinants are the volume
fraction of red blood cells, plasma viscosity,
fibrinogen and lipoprotein (Lowe et al., 1997).
Studies report that for every kilogram of
weight loss, high density lipoprotein (HDL)
cholesterol increases by  and
triglycerides decrease by 
(Dattilo and Kris, 1992), Other studies also
indicate that ketogenic diets results in
significant decrease in serum triglycerides,
small increase in total and low density
lipoprotein (LDL) cholesterol and a moderate
increase in high density lipoprotein (HDL)
cholesterol in subjects with normal lipid
profile (Hossein et al, 2006). Elevated fasting
triglycerides is found to be an independent
risk factor for cardiovascular disease (Austin
et.al,1998), on the other hand numerous
studies suggest that high carbohydrate diet
raise triglycerides levels and reduce high
density lipoprotein (HDL) cholesterol along
with insulin resistance (Taubes, 2001; Zammit
et al, 2001) and when carbohydrate was
replaced with saturated fat the changes in
triglycerides were evident, where high density
lipoprotein (HDL) cholesterol levels increased
drastically when compared to carbohydrate
diet (Reaven, 2000; Krauss, 2001).
Very high levels of ketones make the
blood more acidic and overburden the
kidneys. One of the side effects of a ketogenic
diet is the formation of kidney stones. In
processing higher amounts of proteins, the
kidneys work hard and are forced to excrete
more sodium, calcium and potassium, as well
as filter more of the bile products of protein
metabolism. This extra fluid and electrolyte
loss can cause low blood pressure, which is
another function mediated by the kidneys.
Ketosis in the presence of diabetes can lead to
‘Ketoacidosis’ and coma and can be life
threatening.
Studies have shown that changing of
diet from high carbohydrate low fat diet to
high fat low carbohydrate diet resulted in a
significant reduction in body weight,
improved glycemic control and also led to a
reversal of a six years long decline of renal
function in patients with type 2 diabetes,
(Jorgen et al., 2006). Electrolytes consumed
by most people on a ketogenic diet is
insufficient, some of the fatigue which was
demonstrated in early ketogenic diet studies
have been associated with insufficient mineral
intake, especially sodium (Reid and Ugwu,
1987). A known effect of ketogenic diet is a
decrease in blood pressure, most likely due to
sodium excretion and water loss. In
individuals with high blood pressure
(hypertension), this may be beneficial.
Individuals with normal blood pressure may
suffer from ‘orthostatic hypotension’, which is
a light-headedness which occurs when moving
from a sitting to standing posture (De Haven
et al., 1980). The inclusion of sufficient
minerals appears to be able to prevent
symptoms of fatigue, nausea and hypotension
(Reid and Ugwu, 1987). To counteract the
excretion of minerals on a ketogenic diet,
additional mineral intake is required. An
excessive intake of any single mineral
(especially potassium) can be just as
dangerous as a deficiency.
This study was therefore aimed at
determining the effect of the consumption of
this high fat, low carbohydrate diet on plasma

E. B. OMOZEE and O. J. OSAMUYIMEN / Int. J. Biol. Chem. Sci. 12(6): 2703-2712, 2018
2705
viscosity, lipid profile, serum and urine
creatinine, urea, electrolytes and urinary pH
with a view to ascertain its safety on renal and
cardiovascular system.
MATERIALS AND METHODS
Experimental design
Forty (40) albino Wistar rats of either
sex were obtained from the pharmacology
animal house, faculty of pharmacy university
of Benin. The rats were also housed in the
same pharmacology animal house for the
duration of the study. The rats were weighed
on arrival and then acclimatized for a period
of two week. During this period they were fed
on the normal rat chow having free access to
feeds and water. At the end of acclimatization
the animals were grouped into two, group A
(20) experimental group and group B (20)
control group. The experimental group was
fed with 65% saturated fat ketogenic diet,
while the control group was fed the normal rat
chow. Both groups were on the diet for 30
days and they were allowed free access to
feeds and water during the duration of the
experiment.
Collection of sample
Twenty-four hours fasted blood and
urine samples were collected for electrolytes,
and serum creatinine analysis as well as lipid
profile and blood viscosity analysis. The rats
were put in metabolic cages for collection of
24 hour urine sample. The rats were put to
sleep using chloroform in a closed chamber.
Blood samples were collected from the
abdominal aorta and cardiac puncture into
plain tubes and EDTA container.
Biochemical analysis
All the biochemical analysis was done
spectrophotometrically using Randox reagent
kit. Plasma viscosity was measured by Reid
and Ugwu method.
Diet formulation
The experimental group was fed the
65% fatty diet for a period of 3 weeks.
Determination of plasma viscosity
Reid and Ugwu (1987) method was
used to determine the relative plasma
viscosity.
Principle:
This procedure is carried out based on
comparison of flow rate of plasma and
distilled water under equal pressure and
constant temperature and result expressed as
plasma viscosity of plasma relative to that of
water.
Determination of lipid profile
Total Serum Cholesterol was analyzed
using the method of Allain (1974) while HDL
was analyzed using the method of Lopes
(1977). Serum triglyceride was determined by
the enzymatic method of Stein, (1987). LDL
was extrapolated from total cholesterol,
triglyceride and HDL by the method of
Friedwald (1972). The estimation of
electrolytes was carried out using an ion-
selective electrode 4000 (ISE 4000)
(Külpmann and Wien, 1992). Creatinine was
assayed by two point’s kinetic, modified Jafee
increasing reaction by Bartels and Bolumer
(1972). Serum and urine urea were determined
by Berthelot method (Newman and Price
2001). Urine pH was determined using
reagent strip by Free et al. (1957).
All biochemical assays were carried
out in the Clinical Chemistry laboratory of
University of Benin Teaching Hospital. Blood
viscosity was carried in Physiology
Laboratory, University of Benin.
Statistical analysis
Data are presented as mean ± standard
error of mean (SEM). Student t test was used
for the comparison between experimental and
control groups. P<0.05 was considered
statistically significance. The Graph pad prism
version 5.0 statistical package was used for
the result analysis.
RESULTS
Table 2 compares the mean values of
the lipid profile and plasma viscosity
following treatment with ketogenic diet in
Wistar rats.

E. B. OMOZEE and O. J. OSAMUYIMEN / Int. J. Biol. Chem. Sci. 12(6): 2703-2712, 2018
2706
There was a significant (P<0.05)
reduction in total cholesterol concentration of
experimental when compared with the control
group. Plasma viscosity, HDL cholesterol and
triglycerides values did not show any
significant difference when control groups
were compared with the experimental groups.
However, there was a significant reduction in
concentrations of LDL cholesterol
concentration of experimental groups when
compared with the control groups
Table 3 compares the mean values of
serum electrolytes, creatinine, and urea and
body weight following treatment with
ketogenic diet on Wistar rats.
There was a significant (P<0.05)
increase in serum creatinine concentrations of
the experimental group when compared with
the control group. However, values of serum
sodium, chloride, potassium and bicarbonate
did not show any significant difference when
experimental groups were compared with the
control group. There was a significant
(P<0.05) increase in the initial weight of the
control group when compared the final
weight, while the initial weight of the
experimental group did not show any
significant difference with the final weight.
Table 4 compares the mean values of
urine electrolytes, creatinine, urea and urine
pH following treatment with ketogenic diet on
Wistar rats.
There was a significant (P<0.05)
increase in urinary excretion of sodium,
bicarbonate, chloride and creatinine in the
experimental groups when compared with
control group. Our results also showed a
significant (P<0.05) increase in urinary pH of
the experimental group when it was compared
with the control group. There was however no
significant difference in urinary excretion of
potassium and urea when control and
experimental values were compared.
Table 1: Percent composition of experimental diet.
Table 2: The mean values of the lipid profile and plasma viscosity following
Treatment with ketogenic diet in Wistar rats.
Lipid Profile
Control
Experimental
P-Values
Total cholesterol
(mg/dl)
132.8 ± 10.77
95.60 ± 2.64*
0.0006
Triglyceride (mg/dl)
88.20 ± 17.62
94.90 ± 4.67
0.6337
Constituents
Control
(B)
Experimental
(A)
Coconut Flour
-
15
Maize
70
-
Butter
10
65
Fish Meal
15
15
Bone Meal
2
2
Limestone
1
1
Salt
0.5
0.5
Vit-Min Premix
0.5
0.5
Lysine
0.3
0.3
Meth &Cys
0.3
0.3

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HDL (mg/dl)
36.20 ± 2.33
33.20 ± 1.66
0.3146
LDL (mg/dl)
69.00 ± 10.80
43.60 ± 2.16*
0.0072
Plasma Viscosity
1.478 ± 0.064
1.426 ± 0.054
0.5712
*P<0.05 indicates significant difference when control is compared with experimental group.
Table 3: The mean values of serum electrolytes, creatinine, urea and body weight following
treatment with ketogenic diet on Wistar rats.
Parameters
Control
Experimental
P-values
Serum sodium
(mMol/L)
143.2 ± 1.655
143.4 ± 1.077
0.9185
Serum potassium
(mMol/L)
8.160 ± 0.628
7.970 ± 0.331
0.7712
Plasma bicarbonate
(mMol/L)
22.20 ± 0.860
23.20 ± 0.489
0.2950
Serum chloride ion
(mMol/L)
109.6 ± 1.470
110.6 ± 0.792
0.5210
Plasma urea (mg/dl)
48.80 ± 2.67
57.10 ± 4.476
0.2374
Initial Weight (g)
135.2 ± 5.115
170.6 ± 11.95
<0.0001
Final weight (g)
151.2 ± 5.383
170.6 ± 11.21
<0.05
*P<0.05 indicates significant difference when control is compared with experimental group.
Table 4: The mean values of urine electrolytes, creatinine, urea and urine pH following treatment
with ketogenic diet on Wistar rats.
Parameters
Control
Experimental
P-Values
Urine sodium (mMol/L)
35.60 ± 18.92
147.3 ± 23.60*
0.0089
Urine potassium (mMol/L)
36.24 ± 18.29
59.55 ± 8.058
0.1937
Urine bicarbonate (mMol/L)
800.0 ± 184.4
1570 ± 106.5*
0.0019
Urine creatinine (mg/dl)
1.700 ± 0.230
5.470 ± 0.424*
< 0.0001
Urine chloride (mMol/L)
30.00 ± 17.05
136.0 ± 23.97*
0.0122
Urine Urea (mg/dl)
264.6 ± 7.76
265.7 ± 7.99
0.9321
Urine pH
6.800 ± 0.374
7.600 ± 0.163*
0.0380
*P<0.05 indicates significant difference when control is compared with experimental group.

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DISCUSSION
In this study, there was a significant
reduction in total cholesterol concentrations
level in rats fed with the high fat ketogenic
diet when compared with the control group.
This is in agreement with the findings of Paoli
et al. (2010), and Heisset al. (2002) who
reported a significant reduction in total
cholesterol concentration and an increase in
high density lipoprotein cholesterol (HDL-C)
levels. There was however no significant
difference in the high density lipoprotein
cholesterol (HDL-C) levels between the
experimental group and the control group
when compared. Elevation of HDL-C
concentration is very vital and helps in the
reduction of coronary heart diseases (Eric et
al., 2005 and Paoli et al., 2013).
There was no significant difference in
the triglyceride level between the
experimental group and the control group
when compared. The low-carbohydrate
ketogenic diet has also shown to particularly
affect the level of blood triglycerides
(Hussein et al., 2004; Borge et al., 2014).
There are also significant positive effects on
total cholesterol reduction and increase in
high- density lipoprotein (Hussein et al., 2004;
Hossein et al., 2006; Paoli et al., 2010).
Increase in triglyceride concentrations
is strongly associated with low concentration
of HDL-C. Triglyceride forms a major
component of the skin oil (Paoli et al., 2013);
hence a longer time might be required for any
factor that can influence the body triglyceride
concentration to affect blood parameters.
Findings in this study showed a
significant reduction in low density
lipoprotein cholesterol (LDL-C) levels of the
experimental group when compared with the
control group. This is in line with the findings
of Eric et al., (2005), who observed a
significant reduction in low density
lipoprotein particle (LDL-P) concentrations
after feeding their subjects with high fat
ketogenic diet. In their study they observed a
greater increase in large LDL and greater
reduction in medium LDL. The reduction in
LDL-C observed in our study in rats fed the
ketogenic diet shows that the use of this diet
can be favorable to individuals who have
cardiovascular diseases since this diet tend to
reduce LDL-C, elevation of LDL-C
predisposes an individual to cardiovascular
disease (Borge et al., 2014).
Values of plasma viscosity of control
and experimental did not show any significant
difference. The fact that the plasma viscosity
of the experimental group did not increase
even with the high fatty diet can be attributed
to the values of LDL cholesterol which
decreased in the experimental group. A
relationship between LDL cholesterol and
plasma viscosity has been reported by James
and Robert (1998), according to their findings,
a decrease in LDL cholesterol leads to a
decrease in fibrinogen concentration as a
result a decrease in plasma fibrinogen
concentration which is a major determinant of
plasma viscosity.
There was also a significant reduction
in the weight of rats on the ketogenic diet after
about 2 weeks intake of the diet, This is in
agreement with the findings of Hossein et al.
(2006) who reported a decrease in weight and
corresponding reduction of total lipid profile
but increase in HDL cholesterol. Westerp
(2009), also reported weight reduction after
intake of high fat ketogenic diet and attributed
the reduction in weight to a decrease in calorie
intake due to increased satiety level. Paoli
(2013) also reported a reduction in weight;
however the mechanism of weight reduction
remains unclear.
Serum electrolyte concentrations
showed no significant change in sodium,
potassium, chloride and bicarbonate ions
when compared with the control group. This
is in agreement with the work of Henrietta and
Olumese (2010), who reported no significant
difference in serum electrolyte level after the
intake of ketogenic diet.
In this study, we observed a significant
increase in serum creatinine concentrations of
the experimental group when compared with
the control group. Serum creatinine
concentration is a useful determinant in the
determination of renal disease, according to
Sidney and Shreeran (1997), an increase in
serum creatinine level from the first value to a

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2709
high value is an indicator of underlying renal
failure.
Creatinine is a breakdown product of
creatine phosphate in muscles, and is usually
produced at a fairly constant rate by the body
depending on muscle mass (Yuegang and
Chengjun, 2008). However, in this study, the
experimental groups were not adding weight
probably building muscles hence, the increase
in serum creatinine levels. The increase in
serum creatinine observed in this study could
be attributed to an increase in muscle mass.
These findings are in accordance with the
findings of Alessandra et al., (2008), who
reported that creatinine excretion is also
influenced by muscle mass, because creatinine
formation occurs almost exclusively in the
muscle. These findings further corroborate
that intense physical activity directly
influences body composition, reducing the
body fat and increasing the fat-free mass. This
also agrees with the findings of Sapna et al.
(2012), who reported that in both health and
disease, under steady state, serum creatinine
can serve as a reliable muscle mass biomarker
if appropriate adjustment for full or residual
kidney function and dietary meat intake is
undertaken.
The result from this study also showed
that urinary electrolyte concentration showed
a significant increase in sodium, bicarbonate
and chloride ions when compared with control
and no significant difference in potassium
when compared with control. Electrolytes are
charged particles in body fluids that helps
transmit electrical impulses for proper nerve,
heart and muscle impulses (Keeet al., 2004).
Sodium and chloride, the components of table
salt, are mostly concentrated outside the cells,
while potassium is more concentrated inside
the cells. The balance of these electrolytes on
either side of the cell membrane not only
allows electrical currents to pass between cells
but also helps maintain the proper fluid
balance in the tissues. Studies have shown that
electrolytes consumed by most people on
ketogenic diet is insufficient Phinney (1991)
reported fatigue which is demonstrated in
early ketogenic diet may be due to insufficient
mineral intake especially sodium.
Urinary creatinine concentration
showed that there was a significant increase
compared to control group. Creatinine is a
nitrogenous waste present in urine. Thus, an
increase in muscle mass will lead to an
increase in creatinine production and
excretion. Urea value showed no significant
change also indicating a normal kidney
function. The urine pH showed a significant
increase in the experimental when compared
to control this can be attributed to increased
excretion of bicarbonate in urine; this also
agrees with study by Eiya and Obika (2016),
in their study, they observed a significant
increase in urinary pH after intake of high
casein diet.
Conclusion
Findings from this study has clearly
shown that intake of high fat ketogenic diet is
not harmful to the heart due to the effect of
this diet on LDL and HDL cholesterol as well
as total cholesterol. Even though we did not
observe any significant reduction in
triglyceride values, the fact that there was no
significant increase between triglyceride
values of control and the experimental group
is something worth noting, knowing that
triglyceride makes the skin fat and hence
longer time is required before its blood
parameters can be affected. The fact that this
diet did not result in weight gain shows that
this diet can also be protective against most of
the metabolic syndromes since obesity has
been reported to be a predisposing factor to
most metabolic syndromes. The increase in
serum creatinine observed in this study could
be attributed to increase in muscle mass in rats
fed the ketogenic diet. However, there is need
to carry out further study to ascertain the
actual cause of the increase in serum
creatinine.
COMPETING INTERESTS
The authors declare that they have no
competing interests.
AUTHORS’ CONTRIBUTIONS
EBO conceived the idea for the work
and wrote the proposal; OJO assisted in the

E. B. OMOZEE and O. J. OSAMUYIMEN / Int. J. Biol. Chem. Sci. 12(6): 2703-2712, 2018
2710
technical aspect of this study and in editing
the manuscript. Both parties gave final
approval to the manuscript.
ACKNOWLEDGEMENTS
We will like to appreciate members of
staff in the animal house for assisting in
caring for the animals in the course of the
study.
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