Comparison of Purple Passion Juice (Passiflora edulis var. edulis) and

Simvastatin on Lipid-lowering Effect of Hyperlipidemic Rats Model

Alfi Muntafiah

, Tisna Sendy Pratama

, Anisa Rachmawati

, Dewi Wahyu Wulandari

and Qodri Santosa

Department of Biochemistry, Faculty of Medicine, Jenderal Soedirman University, Purwokerto, Indonesia

Department of Child Health, Faculty of Medicine, Jenderal Soedirman University, Purwokerto, Indonesia

Keywords: Purple passion juice, lipid profile, hyperlipidemia, Simvastatin, propylthiouracil, total cholesterol

Abstract: Hyperlipidemia is a lipoprotein metabolic disorder characterized by high cholesterol and triglycerides in blood

circulation. Non-pharmacological management efforts by consuming healthy foods are often the first choice.

In the previous research, we found the antihypercholesterolemia potential of purple passion juice 4.2

mL/200gBB/day in experimental animals. This study aims to compare purple passion juice's effectiveness to

Simvastatin in improving the lipid profile of the hyperlipidemic rats model. This true experimental study used

32 male Wistar rats, divided into 4 groups: K1 (normal control), K2 (hyperlipidemia control), K3 (purple

passion juice 4.2 mL/200gBB/day), K4 (simvastatin 0.18 mg/200gBB/day). We used pork oil six

mL/200gBB/day and Propylthiouracil (PTU) 12.5 mg/day, divided into two doses, for 14 days, by gavage, to

induced hyperlipidemia. Blood sampling through retro-orbital veins was carried out at the end of induction

(pre-test) and the end of the study (post-test) to measure the lipid profile. T-paired test on total cholesterol

and triglyceride levels of the pre-post test showed a significant decrease in total cholesterol levels in K4

(p=0.00) and a significant decrease in triglyceride levels K3 (p=0.03), while in K1, there was a significant

increase in both parameters. HDL levels decreased significantly in all groups. There was a significant

difference in the one-way ANOVA test for both total cholesterol and triglyceride levels (p <0.05). Post Hoc

test on post-test total cholesterol levels showed a significant difference in K2 v.s K4 (0.00), but there was no

difference between K3 v.s K4 (p=0,54). The study concludes that Simvastatin was more effective in reducing

total cholesterol levels; however, purple passion juice is more effective in lowering triglyceride levels.

1 INTRODUCTION

Cholesterol and triglycerides are the main lipid

components in the blood circulation (Harikumar et

al., 2013). Their normal blood circulation levels play

an essential role in normal physiological processes

(Abumrad and Davidson, 2012). Increasing levels of

these lipid compounds in the circulation are known as

hyperlipidemia (Harikumar et al., 2013). Lifestyle

changes and high calorie, saturated fat, and

cholesterol-rich diet cause this condition (Arsana et

al., 2019). The increase in these compounds in the

circulation can lead to the formation of free radicals,

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initiate cellular damage, and play a role in

atherosclerosis's pathophysiological process

(Harikumar et al., 2013; Nghiem-rao, Mavis and

College, 2014). Management of hyperlipidemia

requires a comprehensive strategy: pharmacological

and non-pharmacological therapy. Pharmacological

therapy uses anti-lipid drugs, including Simvastatin,

a statin group, as the first-line drug. On the other

hand, non-pharmacological treatment is often the first

choice through a healthy lifestyle by consuming foods

and fruits high in antioxidants, phytosterols, low

calorie, and rich fibre (Arsana et al., 2019). The

indigenous systems of medicine have provided

148

Muntaﬁah, A., Pratama, T., Rachmawati, A., Wulandari, D. and Santosa, Q.

Comparison of Purple Passion Juice (Passiﬂora edulis var. edulis) and Simvastatin on Lipid-lowering Effect of Hyperlipidemic Rats Model.

DOI: 10.5220/0010489301480154

In Proceedings of the 1st Jenderal Soedirman International Medical Conference in conjunction with the 5th Annual Scientiﬁc Meeting (Temilnas) Consortium of Biomedical Science Indonesia

(JIMC 2020), pages 148-154

ISBN: 978-989-758-499-2

extensive plant data for the treatment of

hyperlipidemia. Various studies on local natural fruits

and plants, among others, have proven their

effectiveness in treating it (Harikumar et al., 2013;

Alam and Mishra, 2017).

Indonesia is a country rich in various tropical

fruits, which are beneficial for health. Although

scientifically not confirmed about its efficacy, the

practice of using this plant material is widely used as

a preventive or first treatment (Barbalho et al., 2012;

Maqbool et al., 2019). This is based on the general

belief that using these natural ingredients is free from

side effects and is readily available in the surrounding

environment (Kaur, Jasvir; Kaur, Satvinder;

Mahajan, 2013). According to the World Health

Organization (WHO), about 80% of the world's

population, especially developing countries, use this

plant to get it (Soares et al., 2012; Alam and Mishra,

2017). In line with the high public interest in the use

of natural ingredients, the scientific community,

through their research and publication, has

demonstrated the beneficial health effects of

consuming this plant due to its phytotherapeutic

properties (Matsui et al., 2010; Barbalho et al., 2012;

Soares et al., 2012).

Purple passion fruit (Passiflora edulis var. Edulis

Sims) is a tropical fruit that has been successfully

developed in many countries in the world, including

Indonesia. This native Brazilian plant, easy to

cultivate and grow in our environment but is often not

used because of its sour taste. In fact, behind the taste,

this tropical fruit is rich in nutritional and

phytochemicals components, making this exotic fruit

has a beneficial health value. Based on research,

purple passion fruit contains fibre, minerals, phenolic

and ascorbic acids (Ramaiya et al., 2019). In

Indonesia, research and publications regarding the

health benefits of this fruit are still limited.

Previous research has studied the potential of

various parts of the passion fruit plant as a herbal

medicine source. These studies included the effect of

the rind on the maintenance of glycemia (Barbalho et

al., 2012), mesocarp in reducing

hypercholesterolemia (Corrêa et al., 2014; Ostrowski

et al., 2015), fruit peel in cases of asthma (Ross et al.,

2008), and also leaf extract as antidiabetic

(Kanakasabapathi and Gopalakrishnan, 2015).

Exploring the potential of passion juice as an

antihypercholesterolemic agent has also been carried

out in previous studies (Damasceno et al., 2011;

Maricelma da Silva Soares de Souza, Sandra Maria

BArbalho, Debora Christina Damasceno, Marilza

Viera Cunha Rudge, Kleber Eduardo de Campos,

2012). In the previous study, we found the

antihypercholesterolemia potential of purple passion

juice (Muntafiah, Ernawati, et al., 2017). Through the

previous research, purple passion juice at a dose of

4.2 mL/200gBB/day significantly reduced blood

cholesterol levels in experimental animals

(Muntafiah, Ernawati, et al., 2017). Continuing the

previous research, still focusing on purple passion

juice, in this study, we wanted to report the

effectiveness of anti-hypercholesterolemia potential

of purple passion juice compared to the standard drug,

Simvastatin.

2 MATERIALS AND METHODS

2.1 Subjects

This true experimental study used 32 white male rats

(Rattus norvegicus) Wistar, 2-3 months old, and 125-

250 gr weight. The animals were obtained from the

Laboratory of Pharmacology and Experimental

Animals at Jenderal Soedirman University,

Purwokerto. The animals were kept in an

environment with a temperature of 22 ± 2 ° C, 50%

humidity, and a light-dark cycle for 12 hours

(Sambodo et al., 2019). They were acclimatized for

seven days, placed in individual cages with the same

material, shape, and size, received AD II pellet and

aquadest ad libitum. They were divided into four (4)

groups: normal control (K1), hypercholesterolemia

control (K2), treatment with purple passion juice 4.2

mL/200gBB/day (K3), and treatment with

Simvastatin 0.18 mg/200gBB/day (K4).

2.2 Hypercholesterolemia Induction

Induction of hypercholesterolemia was carried out

after the acclimatization period to the K2, K3, and K4

groups. We used pork oil six mL/200gBB/day and

propylthiouracil (PTU) 12.5 mg/day dissolved in 1

mL of water, divided into two doses, by gavage.

Induction was carried out for 14 days (Kusumastuty,

2014; Muntafiah, Ernawati, et al., 2017).

2.3 Purple Passion Juice Preparation

Fresh purple passion juice is obtained from Pulisen,

Boyolali, Central Java. Purple passion fruit is washed,

cut into two parts, and the pulp is taken, then squeezed

with a filter cloth to separate the juice from the seeds.

Making purple passion juice is done every day to

obtain fresh juice (Muntafiah, Pratama and Ati,

2019).

Comparison of Purple Passion Juice (Passiﬂora edulis var. edulis) and Simvastatin on Lipid-lowering Effect of Hyperlipidemic Rats Model

149

2.4 Blood Sampling

Blood samples were collected through the retro-

orbital vein using a hematocrit pipette. Before

sampling, the animals have fasted for 10 hours.

During this time, the experimental animals were still

given aqua dest ad libitum. Blood sampling was

carried out twice, a pre-test (after induction of

hypercholesterolemia) and post-test (end of research).

Furthermore, the sample was inserted into a non-

EDTA tube, centrifuged, and serum was taken.

2.5 Variable Measurement

Examination of lipid profile (total cholesterol,

triglycerides, HDL) was carried out at Biochemistry

Laboratory, Faculty of Medicine UNSOED,

Purwokerto. Total cholesterol levels were examined

using the CHOD-PAP method, using the Dyasis kit.

Triglyceride levels were analysed using Glycerol-3

Phosphate-Oxidase (GPO) method, while HDL levels

were analysed using the CHOD-PAP method. The

sample was examined with a photometer 5010

(Robert Riele GmbH & Co KG, Germany) at a

wavelength of 540 nm (Muntafiah, Ernawati, et al.,

2017).

2.6 Data Analysis

Data analysis was carried out quantitatively. Data

distribution was tested with Shapiro Wilk, and

Levene tested the homogeneity of the data. A paired

t-test was conducted to compare the mean of the pre-

post test of total cholesterol, triglyceride, and HDL

levels in each group—the One Way Anova test was

conducted to compare the mean levels of parameters

tested between groups. One Way ANOVA test was

significant if it was obtained p <0.05 and continued

with post Hoc LSD.

3 RESULTS

3.1 Animal Weight

Weight measurements were carried out five times

during the research period. The first measurement

was carried out at the beginning of acclimatization,

whereas the second was carried out before the

induction. The third measurement was then filled

after hypercholesterolemia induction/before

treatment followed by the fourth measurement, which

carried out in the first week of treatment. The final

measurement was taken at the end of the research.

The average body weight of experimental animals

during the research period is presented in Figure 1.

Figure 1. Bodyweight of the Mice.

Table 1. Lipid profile of the animal model.

Lipid profile

Groups

K1 K2 K3 K4

Total cholesterol (mg/dL)

pre-test 67.5 ± 7.42

125 ± 19.35

104.67 ± 20.32

133 ± 29.79

post-test 113.00 ± 21.79

110.5 ± 16.82

84.17 ± 16.12

79.67 ± 11.38

delta* 45.5 -14.4 -20.50 -53.33

p pre-post ** 0.00 0.20 0.05 0.00

Triglycerides (mg/dL)

pre-test 61.67 ± 18.81

215 ± 88.95

261 ± 96.17

176.67 ± 102.83

post-test 186.83 ± 70.24

161.17 ± 34.44

139.67 ± 28.92

157.33 ± 45.89

delta* 125.16 -53.83 -121.33 -19,34

p pre-post ** 0,02 0,25 0,03 0,6

HDL (mg/dL)

pre-test 85.33 ± 41.10

109.33 ± 24.41

79.67 ± 23.13

82.17 ± 14.46

post-test 46.17 ± 8.57

51.33 ± 13.49

48.67 ± 9.22

37.5 ± 9.29

delta* -39.16 -58 -31 -44.67

p pre-post ** 0,054 0,00 0,03 0,00

* Delta (the mean difference between pre-test and post-test, positive indicates an increase, negative indicates a decrease.

** p<0,05 in each column shows a significant difference between pre-test and post-test.

a,b

Different notations on the same line indicate significant differences.

100

200

300

12345

weight(gr)

weighingtime

JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )

150

3.2 Lipid Profile

Based on Table 1. we can see that the normal control

group (K1) had a mean total cholesterol level of 67.5

mg/dL, triglycerides 61.67 mg/dL, and HDL 85.33

mg/dL. Hyperlipidemia induction using pork oil and

propylthiouracil in the K2, K3, and K4 groups,

significantly increased the total cholesterol and

triglyceride levels. T-paired test on pre-post test total

cholesterol levels showed a significant decrease in K4

groups. On the other hand, in the normal control

group, there was a considerable increase. Likewise,

for triglyceride levels, the paired t-test showed a

significant decrease in triglyceride levels only in K3,

while in K1, there was a considerable increase.

Meanwhile, HDL levels decreased significantly in all

groups. One Way ANOVA test was carried out on

pre-test total cholesterol and triglyceride levels (after

hypercholesterolemia induction) showed a significant

difference in the mean levels of total cholesterol and

triglycerides (p <0.05). Furthermore, the post hoc test

results showed a significant difference between K1

and K2, K3 and K4 (this indicates the success of

hypercholesterolemia induction). Furthermore, one

Way ANOVA test for total cholesterol and

triglyceride levels showed a significant difference (p

<0.05). Post Hoc test on cholesterol levels showed a

significant difference (p <0.05) in K2 v.s K4.

However, there was no difference between K3 v.s K4

(p = 0.54).

4 DISCUSSION

A high-fat diet is directly linked to hyperlipidemia in

humans. To test the herbal effect on reducing

circulating lipids, an experimental study was carried

out with an experimental animal model of

hyperlipidemia in the laboratory (Leite Matos et al.,

2005). Animal models used to identify herbal

remedies' effectiveness can mimic human

pathophysiology (Briggs et al., 2014). In this study,

we made an experimental animal model of

hyperlipidemia. Hyperlipidemia induction was done

by giving pork oil six mL/200gBB/day (Kusumastuty,

2014) and propylthiouracil (PTU) 12.5 mg/day for 14

days (Untari and Pramukantoro, 2020) by gavage.

This induction technique can increase total

cholesterol and triglyceride levels in Wistar rats.

Based on this research, in the regular control group,

the mean total cholesterol was 67.5 mg/dL, and the

average triglyceride level was 61.67 mg/dL.

According to the reference, this average level states

that the range of normal total cholesterol levels for

experimental animals rats aged 8-16 weeks is 37-85

mg/dL and triglycerides 20-114 mg/dL (Giknis and

Clifford, 2008). The results of examining total

cholesterol levels in the normal group were not much

different from previous studies (Muntafiah, Yulianti,

et al., 2017; Muntafiah, Ernawati, et al., 2017).

Meanwhile, in the induced group (K2, K3, K4), total

cholesterol levels were >110 mg/dL, and triglycerides

were >170 mg/dL. The hyperlipidemia induction

technique can increase total cholesterol and

triglyceride levels higher than the previous study. It

was three mL/200gBB/day of pork oil and two

mL/200gBB/day duck egg yolk for ten days

(Muntafiah, Ernawati, et al., 2017. Another one was

giving pure cholesterol of 0.03 grams and 1 gram of

pork oil for 28 days (Sambodo et al., 2019). All the

administration has not propylthiouracil 12.5 mg/day,

and a high-fat feed mixture of quail egg yolk and

ducks egg yolk for 21 days (Untari and Pramukantoro,

2020). Pork oil contains about 38-43% saturated fatty

acids and cholesterol. Pork oil continuously for 14

days resulted in increased cholesterol and triglyceride

levels, accompanied by increased lipoproteins

(hyperlipoproteinemia) in the blood. (Kusumastuty,

2014). Meanwhile, PTU can inhibit thyroid cells in

experimental animals to inhibit thyroid hormone

production and results in hyperthyroidism (Kurniati

et al., 2018). This condition directly affects

lipoprotein metabolism, increasing cholesterol levels

(Untari and Pramukantoro, 2020). Propylthiouracil

(PTU) is a thyroid hormone antagonist. Under normal

circumstances, the thyroid hormone can increase fat

metabolism. PTU is an antithyroid substance that can

inhibit the formation of thyroid hormones that play a

role in lipolysis so that inhibition of this thyroid

increases blood cholesterol concentrations by

expanding endogenous cholesterol biosynthesis

(Diah et al., 2012; Kurniati et al., 2018).

Based on this study, at the end of the research

period, a significant increase in total cholesterol and

triglyceride levels occurred in K1 as a standard

control. Various factors, including dietary factors, can

cause this. In this study, experimental animal feed

with AD II pellets, the fat content in it at least 4%, is

thought to be one factor that causes an increase in

total cholesterol and triglyceride levels. Meanwhile,

in the K3 group, giving purple passion juice 4.2

mL/200gBB/day for 14 days can reduced total

cholesterol and triglyceride levels (Table 1). Previous

research by Soares et al. showed the same thing where

P.edulis 1,000 mg/kg given twice a day for 28 days

showed the effect of improving lipid profile by

increasing HDL levels, lowering total cholesterol

levels, and improving lipid peroxidation in Wistar's

Comparison of Purple Passion Juice (Passiﬂora edulis var. edulis) and Simvastatin on Lipid-lowering Effect of Hyperlipidemic Rats Model

151

Rat (Soares et al., 2012). Something different, in our

study, purple passion juice can reduce triglyceride

levels significantly.

What components are contained in purple passion

juice so that they can reduce total cholesterol and

triglyceride levels? Medical plant research generally

carries out on unrefined plant extract materials that

have various elements. It is said that the bioactive

compounds contained therein can work together

synergistically to produce an effect (Moreira et al.,

2014; Briggs et al., 2014). Likewise, purple passion

juice's antihyperlipidemic potency can be caused by

various bioactive compounds contained in it. The

essential nutrients needed by the body are found in

this juice. This fruit juice contains much fibre, 2.81%

protein, 6.57% carbohydrates, and <0.5% fat content.

In 247 mL (1 cup) of juice contains 24% K, 60-80%

Mg,> 80% P, and 90% Fe of the recommended

dietary allowance of minerals. It has 1137 mg / 100 g

of citric acid (Ramaiya et al., 2019). In addition to

nutritional components, various literature through

their research state that purple passion fruit contains

multiple phytochemical compounds such as

polyphenols, carotenoids, and vitamins (Matsui et al.,

2010) insoluble fibre fraction which is beneficial for

health. Purple passion fruit contains 1,060%

flavonoids, 1.160% carotenoids, and 0.012%

alkaloids. Flavonoids can lower total cholesterol

levels in the blood (Matsui et al., 2010) (Kusumastuty,

2014) through the same mechanism as Simvastatin, a

standardized and first-line drug in lowering

cholesterol levels, by inhibiting the action of the

enzyme 3-hydroxy 3-methyl glutaric coenzyme A

reductase (HMG Co-A reductase) on cholesterol

synthesis in the liver (McFarland et al., 2014).

Besides, flavonoids can also reduce cholesterol

absorption and increase cholesterol conversion into

bile acids so that cholesterol levels in the blood

decrease (Yunarto et al., 2019). Beta carotene can

reduce cholesterol absorption in the intestines and

increase the excretion of cholesterol through faeces.

This situation causes the liver to make cholesterol in

the blood converted into bile acids to reduce blood

cholesterol (Silva et al., 2013). The content of vitamin

C and fibre in purple passion fruit is also thought to

play an active role in lowering blood cholesterol

levels. Vitamin C works by influencing lipoprotein

lipase activity and carnitine synthesis. Lipoprotein

lipase (LPL) is an enzyme that plays a role in

separating triglycerides from chylomicrons and

VLDL. LPL catalyzes the breakdown of triglycerides

into unesterified fatty acids and brings them to the

adipose tissue for reprocessing and storage as

triglycerides. Fibre content can reduce total

cholesterol levels by inhibiting cholesterol

reabsorption from bile salts in the intestine. This fibre

will undergo fermentation in the intestine and then

turn into short-chain fatty acids. It can increase the

intestinal contents' viscosity so that the bile salts'

cholesterol is excreted in the faeces (Matsui et al.,

2010).

5 CONCLUSION

The provision of purple passion juice 4.2

mL/200gBB/day can improve the lipid profile of

hyperlipidemia animals model by reducing total

cholesterol and triglyceride levels. Simvastatin is

more effective in lowering total cholesterol levels;

however, purple passion juice is more effective in

lowering triglyceride levels. The administration of

purple passion juice and Simvastatin 0.18

mg/200gBB/day both had the same effectiveness in

reducing total cholesterol levels in experimental

animals.

ACKNOWLEDGMENTS

This research was funded by the Institute for

Research and Community Service (LPPM), Jenderal

Soedirman University, Purwokerto, Indonesia,

through BLU grant (Competency Improvement

Scheme).

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JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )

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