The Effects of Passiflora Edulis Sims’s Seed Extract on Total

Cholesterol in Atherogenic Young Rats

Dina Keumala Sari

, Sunna Vyatra Hutagalung

, Marianne

, Sri Lestari

, and Muhammad Haekal

Nutrition Departmen, Faculty of Medicine, Universitas Sumatera Utara, Jl. dr. Mansur Kampus USU Medan 20155

Faculty of Tropical Medicine, Mahidol University, Thailand

Pharmacology Pharmacy Department, Faculty of Pharmacy, Universitas Sumatera Utara, Jl. dr. Mansur Kampus USU

Medan 20155

Public Health Departmen, Faculty of Medicine, Universitas Sumatera Utara, Jl. dr. Mansur Kampus USU Medan 20155

Pre-clerkship Medical Doctor Program, Faculty of Medicine, Universitas Sumatera Utara, Jl. dr. Mansur Kampus USU

Medan 20155

Keywords: Passiflora Edulis Sims’s Seed Extract, Cholesterol, Young Rats.

Abstract: Purple passion fruit (Passiflora edulis Sims) is one of antioxidant that contain flavonoid, as a protection

atherosclerosis process. Its juice has been proof to reduced total cholesterol, but its seed also has been

predicted to reduce it. The objective was to find the effects of Passiflora edulis Sims’s seed extract in

different doses in affecting lipid profile serum level in young rats (Rattus novergicus) in fourteen days.

There were four groups containing 20 male rats which were devided into: normal diet group (C0);

atherogenic diet group (C1); atherogenic diet and 50mg/kg body weight (P1); atherogenic diet and

100mg/kg body weight (P2). Results of this study were there was significant difference of body weight,

total cholesterol, and triglyceride, but not for low and high density lipoprotein serum level. The result

reported that body weight showed significant difference between C0, C1, P1, and P2. Total cholesterol

serum between C0 and C1 group (p=0.04), also between C1 and P2 (p=0.003). There was also significant

difference of triglyceride serum level between C0 and P1 (p=0.04) and C0 and P2 (p= 0.03). Based on these

results it can be concluded that Passiflora edulis Sims’s seed extract has potential effect in lowering total

cholesterol and triglyceride serum in young rats.

1 INTRODUCTION

The high incidence of atherosclerosis diseases,

promotes the research for more effective treatments

and looking for better treatment, one of them is with

antioxidant treatment. However, improvement of

lifestyle especially diet therapy, such as lowering

cholesterol food sources is important for the

prevention of cardiovascular disease (García, 2016;

Larson-Meyer et al., 2006; Matsuzawa et al., 2011).

Mixtures of bioflavonoids in the diet and as

supplements have been recognized to play a

protective role by reducing the prevalence of

cardiovascular diseases, one of them is passion fruit.

It has been used widely in folk medicine in tropical

countries as a hermal treatment to treat anxiety,

insomnia, asthma, bronchitis, and urinary tract

infection. The extracts of different parts of the

passion fruit plant contain flavonoids compounds,

but lack of research found flavonoid in the seed

(Baur et al., 2006).

Passiflora edulis Sims (Passifloraceae) is a

woody climber, commonly native of tropical

country, chiefly for its edible fruits. The plant is

commonly called as yellow passion fruit, maracuja,

yellow granadilia, and pomme liane jaune. In

traditional system of medicine, P. edulis is used as

sedative, antiasthmatic and emetic. P. edulis leaves

are used in the treatment of insomnia and

traditionally known to produce a restful sleep

without any narcotic hangover (Kawakami et al.,

2014; Kershaw & Kim, 2017). All parts of the fruits

is edible, including the leaves, that have been

reported to contain a bitter principle maracugine,

resins, acids and tannin exceptionally rich in

ascorbic acid (Kirkland, 2013; Sunitha & Devaki,

2009). The leaves used to treat epilepsy, ulcers and

haemorrhoids, but need further research for the seed.

708

Sari, D., Hutagalung, S., Marianne, ., Lestari, S. and Haekal, M.

The Effects of Passiﬂora Edulis Sims’s Seed Extract on Total Cholesterol in Atherogenic Young Rats.

DOI: 10.5220/0010085107080712

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

708-712

ISBN: 978-989-758-449-7

Although the antioxidant activities of some of

the purple passion fruit peel extract's compounds

have been studied extensively (Lagouge et al., 2006;

Matsui et al., 2010), but activities of the seed has not

been showed beneficieal effects. No data are

available with respect to the clinical usefulness of

Passiflora edulis Sims seed’s extract in lowering

cholesterol total and triglyceride. This research

hypothesized that the Passiflora edulis Sims seed’s

extract, a novel mixture of bioflavonoids, may

attenuate the lowering cholesterol total and

triglyceride in fourteen days.

2 MATERIAL AND METHOD

2.1 Animal Treatment

The study was conducted in accordance with the

ethical principles for animal experimentation and the

study was approved by the ethics committee on

animal research at the Animal Research in Biology

Faculty of Universitas Sumatera Utara, with ethical

number 112/KEPH-FMIPA/2018. In total, 25 young

male Wistar rats (Rattus norvegicus) weighing 180-

250 g that were obtained from the animal research

were used in this study. The rats were housed in a

temperature-controlled room on a 12 h light/dark

schedule with food and water available ad libitum.

All the young rats devided into four groups,

which were normal diet group (C0); atherogenic diet

group (C1); atherogenic diet and 50mg/kg body

weight (P1); atherogenic diet and 100mg/kg body

weight (P2). Normal diet group given normal food

pellet and for atherogenic diet gave atherogenic food

pellet. The ethanolic seed’s extract was diluted in

water and administered to the animals by gavage for

two weeks, after the induction of atherogenic.The

animals were anaesthetised on the 14th day, and

blood was collected by cardiac puncture after a 12-

hour fast. The animals were euthanised, and

collected for blood serum.

2.2 Passiflora edulis Sims’s Seed

Extract

The purple passion fruit (Passiflora edulis f edulis

Sims) belongs to the genus Passiflora of the family

Passifloraceae. It is grown in Sumatera Island, found

in Berastagi Farm, North Sumatera, Indonesia. The

Passiflora edulis Sims seed were dried at 40 °C,

powdered in an industrial blender, extracted by 70%

ethanol maceration, evaporated under reduced

pressure and lyophilised.

2.3 Flavonoid Analysis

The Passiflora edulis Sims’s seed extract was

analyzed by high-performance liquid chroma-

tography (HPLC) on a Hewlett-Packard 1100

instrument equipped with a DAD detector and a

LiChrospher 100 RP-18 column (5 μm), 125 × 4 mm

held at 30°C, as described elsewhere. The solvent

program started from 3.6% B (2% HOAc in

acetonitrile) in solvent A (2% HOAc in water) up to

12% B in 20 minutes, to 20% in 30 minutes, and to

50% B in 45 minutes. Flow rate was set at 1 mL/min

and compounds were monitored by UV absorption

set at 280 nm for phenolic acids, 350 nm for

flavonoids, and 520 nm for anthocyanins. The

identity of the compounds was confirmed by

comparison of their retention time and UV/ visible

spectra with those of authentic materials

(Rodrigues).

2.4 Total Cholesterol Analysis

Cholesterol is measured enzymatically in serum or

plasma in a series of coupled reactions that

hydrolyze cholesteryl esters and oxidize the 3-OH

group of cholesterol. One of the reaction byproducts,

H2O2 is measured quantitatively in a peroxidase

catalyzed reaction that produces a color. Absorbance

is measured at 500 nm. The color intensity is

proportional to cholesterol concentration.

2.5 Low Density Lipoprotein and

High-density Lipoprotein Analysis

A polyclonal antibody specific for LDL has been

pre-coated onto a 96-well microplate with

removable strips. LDL in standards and samples is

competed with a biotinylated LDL sandwiched by

the immobilized antibody and streptavidin-

peroxidase conjugate. All unbound material is then

washed away and a peroxidase enzyme. Incubated in

twelve minutes for LDL and fifteen minutes for

HDL, absorbance is measured at 450 nm.

The Effects of Passiﬂora Edulis Sims’s Seed Extract on Total Cholesterol in Atherogenic Young Rats

709

2.6 Triglyceride Analysis

Triglycerides are measured enzymatically in serum

or plasma using a series of coupled reactions in

which triglycerides are hydrolyzed to produce

glycerol. Glycerol is then oxidized using glycerol

oxidase, and H2O2, one of the reaction products, is

measured as described above for cholesterol.

Absorbance is measured at 500 nm.

2.7 Statistical Analysis

The results were expressed as the mean ± standard

deviation; they were also submitted to one-way

analysis of variance and compared using the LSD

analysis

3 RESULT

The results will be discussed in three subsections,

they were flavonoid analysis, total cholesterol, and

triglyceride serum level of the young rats. Low and

high density lipoprotein did not show significant

difference in all groups, but showed lowering (LDL)

and increasing (HDL) serum level, probably because

of short experimental duration.

3.1 Flavonoid Analysis

In this flavonoid analysis, IC50 value was

calculated, ascorbic acid was used as the reference

standard. The reducing power of the ethanol extract

was carried out (Oyaizu, n.d.). Different

concentrations of the extracts (1000, 500, 250, 125,

62.5, 31.25 μg/ml) were prepared. To all the test

tubes 2.5 ml of sodium phosphate buffer followed by

2.5 ml of 1% potassium ferrocyanide solution was

added. The contents were vortexed well and then

incubated at 50° for 20 min. After incubation, 2.5 ml

of 10 % trichloroacetic acid was added to all the

tubes and centrifugation was carried out at 3000 rpm

for 10 min. To 5 ml of the supernatant, 5 ml of

distilled water was added. To this about 1 ml of 1%

ferric chloride was added to each test tube and

incubated at 35° for 10 min. The absorbance was

read at 700 nm. The reducing power of the extract

was linearly proportional to the concentration of the

sample. Ascorbic acid was taken as reference

standard (Oyaizu, n.d.).

The result of phytochemical analysis was

recorded in Table 1, showed that Passiflora edulis

Sims’s extract containing flavonoid, Tannin, and

Saponin. By comparing with vitamin C standart,

Passiflora edulis Sims’s extract showed strong

antioxidant.

Table 1. Analysis of Flavonoid Content

Sample

Identity

and

sample

Parameter

Results

Analysis Technique

Passiflora

edulis

Sims’s

seed

extract

Density

Phytochemical :

Flavonoid Positive -

Colour

visualisation

Alkaloid

Wagner Negative -

Mayer Negative -

Dragendrof Negative -

Tannin Positive -

Saponin Positive -

Quinon Negative -

Steroid Negative -

Triterpenoid Positif -

Antioxidant IC

DPPH

< 31.25 ppm Spektrophotometry

Vitamin

standard

Density Antioxidant IC

DPPH

6.75 ppm Spektrophotometry

ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches

710

3.2 Total Cholesterol Analysis

The cholesterol total serum level showed lower

serum level between C0 and C1, this may indicate

that atherogenic food pellet for young rats gave

atherogenic effect. Atherogenic food pellet were

high in isaturated fatty acids, cholesterol, and

calorie. There was significant difference between C0

(mean±standart deviation/SD: 84.5±13.7mg/dL) and

P2 (mean±SD: 68.1±6.2mg/dL), it showed that

100mg/bodyweight Passiflora edulis Sims’s seed

extract had lowering effect to total cholesterol serum

level.

Previous research showed that Passiflora edulis

Sims’s seed extract had phytochemical action as

antioxidant (Sun et al., 2007; Szkudelski &

Szkudelska, 2015; Timmers et al., 2011). The

research demonstrated that Passiflora edulis Sims’s

seed extract could lowered blood glucose levels in

mice consuming a high-fat diet. Other previous

study showed anti-hypertensi effect and anti-ageing

effect in rats (da Silva et al., 2013; Fusco & Pani,

2013; Pearson et al., 2008; Tchkonia et al., 2010).

Table 2. Analysis of Total Cholesterol Serum Level

Group Groups Mean

difference

Std

erro

Significance

C0 C1

11.14

10.03

16.49

5.20

4.98

0.44

0.68

0.03

C1 C0

–11.14

–1.11

5.36

5.20

4.98

0.44

0.83

0.29

P1 C0

1.11

–10.03

6.47

5.20

4.98

0.83

0.68

0.21

P2 C0

–5.36

–16.49

–

6.47

4.98

0.29

0.03

0.21

3.3 Triglyceride Analysis

Triglyceride lowering effects showed in group P1

and P2, that would be 50 mg and 100

mg/bodyweight Passiflora edulis Sims’s seed

extract. There was significant difference between C0

(mean±SD: 1.0±0.3mg/dL) and P1 (mean±SD:

0.77±0.3mg/dL). There was also significant

difference between C0 (mean±SD: 1.0±0.3mg/dL)

and P2 (mean±SD: 0.74±0.2mg/dL), it showed that

50 and 100mg/bodyweight Passiflora edulis Sims’s

seed extract had lowering effect to triglyceride

serum level, but based on these result showed that

Passiflora edulis Sims’s seed extract

50mg/bodyweight already showed lowering effect.

Table 3. Analysis of Triglyceride Serum Level

Group Groups Mean

difference

Std error Significance

C0 C1

0.35

0.32

0.35

0.153

0.147

0.04

0.05

0.02

C1 C0

–0.35

–0.02

0.09

0.153

0.147

0.04

0.89

0.95

P1 C0

–0.32

0.02

0.03

0.153

0.147

0.89

0.04

0.83

P2 C0

–0.35

–0.09

–

0.03

0.15

0.95

0.02

0.83

There were several limitations to this study

design, this study occurred over a short time period,

and the number of rats. However, the most effective

dose of Passiflora edulis Sims’s seed extract to be

investigated in future study.

4 CONCLUSIONS

Passiflora edulis Sims’s seed extract has potential

effect in lowering total cholesterol and triglyceride

serum in young rats. This research also concluded

that 100mg/bodyweight Passiflora edulis Sims’s

seed extract had lowering effect on total cholesterol

serum level and 50mg/bodyweight triglyceride

serum level in two weeks in young rats.

ACKNOWLEDGEMENTS

The authors gratefully acknowledge that the present

research is supported by Ministry of Research and

Technology and Higher Education Republic of

Indonesia. The support is under the research grant

TALENTA USU of Year 2018 Contract Number

2590/UN5.1.R/PPM/2018.

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