The Effects of High Fat High Carbohydrate (HFHC) Diet on Body

Weight in Overweight Sprague Dawley Rats

Novia Zuriatun Solehah

, Adi Prayitno

2,3 b

and Eti Poncorini Pamungkasari

2,4 c

Postgraduated Student of Human Nutrition Science, Universitas Sebelas Maret, Surakarta, Indonesia

Postgraduated Program of Nutrition Science, Universitas Sebelas Maret, Surakarta, Indonesia

Departement of Pathology Anatomic, Faculty of Medicine,Universitas Sebelas Maret, Surakarta, Indonesia

Departement of Public Health, Faculty of Medicine, Universitas Sebelas Maret, Surakarta, Indonesia

Keywords: High Fat High Carbohydrate Diet, Body Weight, Overweight.

Abstract: Excessive body weight is caused by an inappropriate diet. Foods that are high in fat and carbohydrates may

lead to overweight. When overweight condition is followed by an increase in the body fat level, it also

increases the production of Reactive Oxygen Species (ROS) in the blood circulation and adipose cells. This

study aims to determine the effect of High Fat High Carbohydrate (HFHC) diet on body weight of overweight

Sprague dawley rats. This study used an experimental design with a pre-post-test control group design. Male

Sprague dawley rats (n=10) were randomized into two groups. The normal group was given standard rats and

the overweight group was given diet HFHC for 14 days. Their body weights were measured before and after

treatment. After 14 days of treatment, the body weight mean was (210 ±1,5) in the treatment group increased

significantly, compared with body weight mean (193±1,3) and before treatment (183,2±1,6) in the control

group. In conclusion, those induced to be overweight used HFHC diet for 14 days and increased body weight.

This finding can be used to induce overweight rats for a short time.

1 INTRODUCTION

Global prevalence of overweight rose trifold (1.9

billion) of world population and around 13.6% in

Indonesia by 2018 (Khadaee and Saidi, 2016; Rebello

et.al., 2020; Riskesdas, 2018). Overweight and

obesity is the fifth leading cause of mortality globally

(Sudargo et.al., 2018). Overweight is a starting point

for obesity which then will lead to various metabolic

syndromes including diabetes mellitus, hypertension,

stroke, coronary heart disease, dyslipidaemia and

cancer (Muhammad et.al., 2019). Results of a study

stated that the increase of Body Mass Index (BMI)

within five years between the age of 25-74 years

significantly increased the risk of hypertension up to

30% compared to people without an increase of BMI

(Hall et.al., 2019). In addition, a rise in body weight

may trigger insulin resistance of peripheral tissue

through disfunction or adipose tissue lipotoxicity,

inflammation, dysfunction of mitochondria,

https://orcid.org/0000-0003-0827-2695

https://orcid.org/0000-0001-5548-4848

https://orcid.org/0000-0002-4197-3226

hyperinsulinemia and endoplasmic reticulum stress

mechanisms (ER) (Longo et.al., 2019).

The accumulation of excess fat causes the

release of high amount of free fatty acid to various

organs thus generating metabolic syndrome. The

prevalence of metabolic syndrome grows as the

prevalence of obesity rises. One of the factors that

sets overweight off is excessive fat and carbohydrate

intake in addition to a sedentary lifestyle. In recent

days, the diet pattern of Indonesians shifts as it

receives heavy influence from western diet pattern

where people consume more high fat and high

carbohydrate food (Fernandez et.al., 2018). Food

with high content of carbohydrate, protein, and fat

may contribute to the increase of oxidative stress and

cause inflammation by forming white adipose tissue

which secretes proinflammation factor (Tan et.al.,

2018; Tan and Norhaizan, 2019). Increased oxidative

stress is the main cause of numerous metabolic

diseases.

106

Solehah, N., Prayitno, A. and Pamungkasari, E.

The Effects of High Fat High Carbohydrate (HFHC) Diet on Body Weight in Overweight Sprague Dawley Rats.

DOI: 10.5220/0010760400003235

In Proceedings of the 3rd International Conference on Social Determinants of Health (ICSDH 2021), pages 106-110

ISBN: 978-989-758-542-5

Imbalanced energy intake and outtake give rise

to excessive fat accumulation not only in adipose

tissue but also in ectopic tissue like the liver. If

prolonged, it will cause excessive fat in liver thus lead

to impaired liver function, fatty liver and liver failure

(Rahman et.al., 2017). The sign of accumulated fat in

ectopic tissue is the increased percentage of visceral

fat in intra-abdominal linked to abdominal obesity

(Tchernof et.al., 2013). Also, excessive fat and

carbohydrate intake may change oxygen metabolism

which then triggers oxidation reaction. Excessive

intake may also induce enlargement of adipose tissue

through adipocyte hypertrophy and hyperplasia. The

imbalance between ROS and antioxidants causes an

increase in oxidative stress, causing systemic

inflammation.

Observing overweight model mice with diet

induction can give illustration of the effect of poor

eating habits among human. Several studies in animal

used high fat high carbohydrate feed in inducing

overweight to animals. This diet is believed to be the

best method in following the pathogenesis of the

development of metabolic syndrome, one of which is

obesity (Panchal et.al., 2011). Although, several

studies conducting the induction of overweight

animal models require quite a long time. Therefore,

this study aimed to examine the effect of the high fat

high carbohydrate (HFHC) diet on overweight rats for

14 days.

2 MANUSCRIPT PREPARATION

2.1 Animal Experimental Protocol

This study has received approval from the Health

Research Ethics Commission, Faculty of Medicine,

Sebelas Maret University Surakarta (KEPK UNS)

No.23/UN27.06.6.1/KEP/EC/2021.

2.2 Samples

The samples in this study were Rattus norvegicus

strain male Sprague dawley rats aged 8-12 week with

the average of weight of 150-200 gram. The subjects

are 10 rats divided into normal or control groups and

HFHC diet group. The subjects were obtained from

the Nutrition Laboratory of the Food and Nutrition

Studies Centre PAU, Gadjah Mada University,

Yogyakarta.

2.3 Study Design

This is an experimental study with pre posttest with

control group design. 10 rats are divided into two

groups, control group and HFHC diet group. Control

group was given standard Comfeed which was AD II

feed. All the rats were acclimatized for seven days in

individual cages and were given standard Comfeed

AD II with ad libitum drinking water. Every 100 gram

of standard AD II feed contains 12% water, 7% ash,

15% crude protein, 3-7% crude fat, 0.9-11% calcium,

0.6-0.9% phosphor, antibiotic, and coccidiostat

maximum 20 mg/h. The raw materials used include

yellow corn, SBM, MBM, CGM Palm olein, essential

amino acid, essential mineral, premix, and vitamin.

The average feed intake was 5 g/100gBB/h. The rat

rearing cage was a plastic cage sized 25 cm x 15 cm

x 7 cm. Each cage is used by only one rat. The

subjects were reared in a specific room with

controlled temperature (27-29ºC) inside a hygienic

polypropylene cage. The room is set with a 12 hours

light and 12 hours dark cycle (lights were turned on

at 07.00 pm, and 70-90% humidity. The rats were

considered stable if the feed consumption and drink

was sufficient. The HFHC group was given HFHC

feed. The composition of the HFHC feed includes 5%

cheese, 10% egg yolk, 15% cow fat, oil 15%, rice

45% and standard feed 20% (Ardiansyah et.al.,

2018). The HFHC diet was given for 14 days. All the

feed was given ad libitum. In a study conducted by

Adriansyah et al, the induction of obese rats was

carried out for 8 weeks. And in this study, HFHC feed

was added with 0.02 g of cholic acid wihch functions

as a fat binder in the body and produce overweight

rats in a shorter time.

2.4 Anthropometry Measurement

The body weight measurement was conducted using

a digital scale. It was a digital scale with 0.1 accuracy.

The measurement was done three times, after 7 days

acclimatization period, after 7 days diet induced

overweight, and after 14 days diet induced

overweight right before termination.

2.5 Statistical Analysis

The statistical analysis was conducted using SPSS

version 16.0. The data was presented with a mean

value and standard deviation. To analyse the mean

difference between groups a paired t test and oneway

anova was performed. The result is considered

significant if the p value is <0.05.

The Effects of High Fat High Carbohydrate (HFHC) Diet on Body Weight in Overweight Sprague Dawley Rats

107

3 RESULT AND DISCUSSION

The development of overweight model rats was

carried out by feeding HFHC. The difference in the

composition of HFHC with the standard can be seen

in Table 1.

Table 1: Composition of Standart and HFHC

redients Standar

HFHC

Carbohydrate 53-57 % 67.69%

Protein 16% 22%

Fat 3-7% 74%

Table 1 shows that the composition of HFHC feed

contains higher carbohydrates and fats compared to

standard feed. The body weight increased due to the

administration of feed with 67.69% carbohydrate and

74% fat from the total calorie. Results displayed that

consumption of fat >30% of total calories can cause

obesity (Hariri and Tibault, 2010). In this study, the

composition of the HFHC feed includes cheese, egg

yolk, cow fat, oil and rice. In addition, the mice

induced with HFHC feed tend to consume a lot of

water. Water is essential in digestive system

especially during the enzymatic hydrolysis and lipid

absorption. Lipase enzyme plays a role to hydrolyse

triglycerides into diglycerides, monoglycerides fatty

acid and glycerol (Udomkasemsab and Prangthip,

2019). The animal response to the diet given to them

can be used to evaluate the rate of success during the

overweight induction. Thus, the composition of

HFHC diet compared to control group’s standard diet

must have a similar basic nutritional content except

for the types of the macronutrients, namely

carbohydrate and fat. For this research, the

composition of macronutrients in HFHC diet include

cheese, cow fat, egg yolk, and rice.

On the other hand, the composition of

macronutrients in control group’s standard feed

included yellow corn as a carbohydrate. The content

of carbohydrate and fat in the HFHC diet is

significantly higher compared to the standard feed

which only contains 53-57% carbohydrate and 3-7%

fat. Results of the systematic review conducted by

Tibault and Hariri (2010) demonstrated that the best

method to induce obesity into mice is by using high

semi-pure fat which contains 40% animal fat with a

low amount of amino acid omega 3 and pure

vegetable oil and high in amino acid omega 6 and

omega 9. The composition of selected raw ingredients

will influence the amount consumed. This will also

affect the increase of the body weight. The

consumption of food high in fat, sugar, and salt

contributes to the occurrence of overweight and

obesity by changing the expression of gene and

sending dopamine signal in the brain. Several

researches mentioned that high fat diet is a source of

fat which in it includes saturated fatty acid (de Moura

e Dias et.al., 2021). Results indicated that there is a

positive association between body adiposity and HFD

including saturated fatty acid (Norris et.al., 2017).

Weighing was carried out to evaluate whether

there was a difference in the weight of rats fed HFHC

for 14 days with rats fed only standard feed. Rats were

weighed 3 times before HFHC induction, after HFHC

induction on the 7 days, and after HFHC induction on

the 14 days. The average of body weight before and

after induction are presented in Table 2.

Table 2: Average on Body Weight Before and After

Induced HFHC

Group

Pre test

(Mean ± SD)

Post test

(Mean ± SD)

Normal 183,2 ± 3,70 193,0 ± 2,92 >0.005

HFHC 184,2 ± 3,77 210,6 ± 3,36 >0.005

>0.005

In table 2 shows that the administration of the

HFHC diet for 14 days significantly increased body

weight (p>0.005), the mean body weight rose from

184,2 ± 3,77 to 210,6 ± 3,36. Within control group

which was given standard feed, the mean body weight

was lower than the HFHC group. This supports the

findings of a study conducted by Wong (2018) which

concluded that HFHC diet for 6 weeks significantly

increased rats body weight to obesity. Other study on

Sprague dawley rats displayed that the administration

of high fat diet of 61% for 8 weeks did develop

obesity and increase visceral fat accumulation

(Udomkasemsab and Prangthip, 2019).

Rats overweight determined based on the Lee

index. The rats stated if overweight index value Lee

> 300. Heavy rats soul uses scales and the height is

measured using ruler. The average of Lee index can

be seen in Tabel 3.

Table 3: Indicator of Overweight

Grou

Post test

(Mean ± SD)

Lee Indekx p

Normal

193,0 ± 2,92 285,97

±1,77

>0.005

HFHC

210,6 ± 3,36 324,76

±4,62

>0.005

Consumption of food high in fat can cause

excessive fat accumulation in our body which then

resulted in the increase of free fatty acid in adipose

ICSDH 2021 - International Conference on Social Determinants of Health

108

tissue and release a high level of triglycoid (Marques

et.al., 2016). Also, excessive fat accumulation will

also be stored in ectopic tissue like liver, muscle, and

heart which will be indicated by hypertrophy and

hypoxia thus lead to inflammation and insulin

resistance (Longo et.al., 2019).

The incidence of overweight and obesity occur

due to the intake of fat that is way higher than the

outtake thus the excess will be stored in the form of

fat in our body, especially adipose tissue. The

increase in adipose tissue mass causes changes in

adipokine production where adipose tissue plays a

role in energy storage which is an important

endocrine. Furthermore, the enlargement of adipose

tissue and the increase of adipocyte progressive may

disturb the blood flow (McArdle et.al., 2013; Exley

MA et.al., 2014). High fat diets have been proven to

increase body weight, fat deposition, increase of

oxidate stress biomarkers, increase of fasting plasma

glucose and insulin. The occurrence of inflammation

around the blood vessel is triggered by the

accumulation of fat in cellular molecules by releasing

inflammation adipocytes like tumour necrosis factor

(TNF) and interleukin-6 (IL-6).

In overweight, the production of adipocytes is

disrupted, including leptin, resistin, adiponectin,

Monocyte Chemoattractant Protein-1 (MCP-1),

Tumor Necrosis Factor-α (TNF-α), and interleukin-6

(IL-6) (Mesquida et.al., 2020). Animal research

conducted by Feillet-Coundray et.al. (2019) showed

that the administration of high fat/high fructose cause

overweight, glucose intolerance, and increase of IL-

6. Analysis results review done by Tan et.al. (2018)

mentioned that high fat diet for 12 months increased

plasma triglycerides and total cholesterol which led to

increase of oxidative stress biomarker in blood. High

fat diet not only worsen lipid profile but also increase

the accumulation of ROS and trigger damage to

mitochondria. The mechanism of inflammation in

adipose tissue is caused by the activation of

proinflammation line in this case being nuclear

factor-kappaB- (NF-κB-). The occurrence of this

inflammation can be detected by the increase

inflammation biomarker and cytokines. In addition,

the occurrence of inflammation is also linked to the

increase of oxidative stress (de Melo et.al., 2017).

Overweight and obese people go through an increase

in the production of raective oxygen species (ROS)

due to imbalance between prooxidant and antioxidant

(Bondia-Pons et.al., 2012; Yosika et.al., 2020). This

condition contributes to the incidence of metabolic

disordersnamely insulin resistance, type 2 diabetes

mellitus, hyperlipidaemia, and atherosclerosis.

4 CONCLUSIONS

Feeding HFHC diet for 14 days can increase body

weight in overweight induced mice. However, further

studies are needed to further analyse the effect of

HFHC to overall health. Furthermore, this study can

be used as a reference to rear overweight model mice

in shorter period of time.

ACKNOWLEDGEMENTS

We would like to thank all staffs in Laboratory of

Food and Nutrition Study Centre of PAU Gadjah

Mada University Yogyakarta for the support in

rearing the rats during this study.

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