The Effectiveness of Pare Extracts (Momordica Charantia L) in

Lowering the Level of Blood Glucose on Wistar Rat (Rattus

Norvegicus) with Hyperglycemia

Dedy Syahrizal

, Ghalia Mayuna

, Teuku Mamfaluti

Department of Biochemistry, Faculty of Medicine Universitas Syiah Kuala, Banda Aceh, Indonesia

Department of Internal Medicine, Faculty of Medicine Universitas Syiah Kuala-Rumah Sakit Umum Daerah Dr. Zainoel

Abidin, Banda Aceh, Indonesia

Keywords: Pare (Momordica charantia L), Hyperglycemia, Blood glucose

Abstract: Pare (Momordica charantia L.) is one of the traditional herbal plants used to cure several diseases, one of

which is to decrease the blood glucose level. This research aims to examine the effectiveness of the ethanol

extract in pare to decrease blood glucose level on male Wistar rat (Rattus norvegicus). This was an

experimental research which used experimental animals with pretest-posttest control group design. The

sample of this research were 40 male mice (Rattus norvegicus)which were divided randomly into 5 groups

of treatment; 2 groups were control groups and 3 groups were given pare extract. All of the male mice were

induced by 10% glucose solution to increase their blood glucose level. In order to examine the response,

the level blood glucose was observed in 30 minutes, 65 minutes, 95 minutes, 125 minutes, and 155 minutes

after the glucose induction. The result of blood glucose level was analyzed using the Anova test and

Duncan test. The result of this study shows that pare extract with 200mg/kgBB group, compared to other

dose level, had faster onset to decrease the blood glucose level of the male Wistar mice that was induced by

10% glucose solution. In conclusion, pare extract effectively decreases the blood glucose level in the

glucose-induced male mice.

1 INTRODUCTION

Diabetes mellitus (DM) is a metabolic disorder. One

of the pathological conditions found in DM is

hyperglycemia. Hyperglycemia leads to various

organs’ disorders. Therefore, diabetes mellitus

usually aims to overcome the hyperglycemic

condition. Some herbal plants are thought to be able

to cure the hyperglycemic condition.

DM treatment can be done in several ways;

i.e. diet (nutritional therapy), insulin, and oral

hypoglycemic drugs (OHO). The long-term usage of

OHO not only requires big budget but also causes

some side effects. The side effects of OHO depend

on the type of the drug and the way it works.

Therefore, people often use herbal remedies that

have not been standardized yet, e.g. pare

(Momordica charantia L).

Previous research has shown that pare extract

can lower the blood glucose levels of alloxan-

induced mice to destruct pancreatic beta cells (type 1

diabetes). However, there is no single study that

examines the direct effects of pare on

hyperglycemia. This study will examine the

effectiveness of pare as anti-hyperglycemic on

glucose-induced mice.

2 RESEARCH METHODOLOGY

This was a laboratory experimental research using

Completely Randomized Design (RAL) with pretest

posttest control group design method. The

maintenance of sample animals and blood sampling

were done at the Faculty of Veterinary Unsyiah; the

Herbarium Test were conducted at Biology

Laboratory; while the pare extract (Momordica

charantia.L.) were processed in the Chemical

Laboratory of Faculty of Math and Natural Sciences

(FMIPA). The study was conducted in September to

October 2017. The sample in this study were wistar

mice (Rattus norvegicus). The Inclusion Criteria are:

Syahrizal, D., Mayuna, G. and Mamfaluti, T.

The Effectiveness of Pare Extracts (Momordica Charantia L) in Lowering the Level of Blood Glucose on Wistar Rat (Rattus Norvegicus) with Hyperglycemia.

DOI: 10.5220/0008791200760080

In Proceedings of the 2nd Syiah Kuala International Conference on Medicine and Health Sciences (SKIC-MHS 2018), pages 76-80

ISBN: 978-989-758-438-1

male; age 3-4 months; body weight 150-200 grams;

healthy condition (active and not disabled); The

Exclusion Criteria: mice have been used for other

studies; and Drop Out Criteria: mice died during the

study. The subject size for each group was measured

using Lameshow formula and got 9 mice per group.

2.1 Research Instruments and

Materials

For maintenance and treatment, there were: cage and

pad, gastric sonde, animal scales, lancet, GlucoDR

glucometer, GlucoDR glucose test strip. For the

manufacture of pare extract, there were: knifes,

blenders, filters, measuring cups, dropper, reaction

tube, tube rack, stir bar, reaction tube shelf, and

vacum rotary evaporator. For the glucose induction,

there was: 1 cc syringe.

For maintenance and treatment, there were:

male wistar mice (Rattus norvegicus), rat blood,

water, and pellets. For the production of pare

extracts, there were: pare (Momordica charantia.L)

and ethanol 70%. For the induction of glucose, there

was: 10% glucose.

2.2 Research Procedures

The Preparation and Maintenance of Samples

Prior to the treatment, the mice were given an

adaptation period for one week, then they were

fasted for 12 hours. During the adaptation process,

their weight and activities were continuously

observed. The mice moved actively and none of

their body weighed less than 200 grams during and

after the adaptation process. Therefore, no samples

were released. Mice were placed in cages that had

previously been dried in the sun to be sterilized and

were given the husk pads. The cage is cleaned and

the husk pad is replaced at least every 3 days.

The mice were fed twice a day and given

aquadest with ad libithum. The pellet used was T79-

4P, with the composition as follows: fish meal,

soybean meal, wheat fraction, rice bran, vitamin A,

vitamin D3, vitamin E, vitamin K, vitamin B2,

vitamin B6, vitamin B12, niacin, calcium D,

panthethonate, choline chloride, minerals, and

antioxidants. The nutrient content of pellets were

protein (16-18%), fat (4%), fiber (8%), ash (12%),

moisture content (12%) (Smith, 1988). The pellet

was given using gastric sonde by inserting a dull

pointed oral needle or cannula into the mouth, then

slowly launched through the ceiling toward the back

until the esophagus into the stomach. During the

adaptation process, the weight and activities of the

mice continuously observed.

Herbarium Test

The herbarium test was conducted at the Research

Laboratory of Department of Biology at Faculty of

Mathematics and Natural Sciences (FMIPA). The

herbarium test was performed to confirm the

taxonomy of pare (Momordica charantia L) and the

following is the taxonomic result:

Kingdom : Plantae

Division : Magnoliophyta

Class : Magnoliopsida

Order : Cucurbitales

Family : Cucurbitaceae

Genus : Momordica

Species : Momordica charantia L

The Preparation and Extraction of Pare

The pare extract was made from 5000 grams of fresh

pares which were crushed using mortal. After that,

the thanol solvent was added, put into the container,

and closed. It was left for two days, isolated from

the sunlight and stirred, then filtered for maserate.

The mixture was macerated using ethanol with the

same procedure, the maceration was performed until

a clear maserate is obtained. Then, all ethanol

maserates were mixed and evaporated using a rotary

vacuum evaporator at a temperature of ± 40 ° C until

a thick ethanolic extract was obtained.

The Determination of Pare Extract Dose

Medical Administration Volume (VAO) is the

volume of dosage administered to animals in ml

units. In a previous study, Adewole et. Al. used

doses at the intervals of 50 mg/kgBW to 400

mg/kgBW. Therefore, it was obtained that the

volume of drug administration (VAO) for pare

extract to be administered is 20 mg/ml, 40 mg/ml,

and 80 mg/ml.

The Determination of Metformin Dose

Human Equivalent Dose (HED) is used to determine

the dose conversion factors used from animals to

humans, using the Body Surface Area (BSA). The

ultimate effect of Metformin are to reduce the

production of liver glucose (gluconeogenesis), and

improve peripheral glucose uptake (Perkeni, 2015).

Metformin has a dose of 1500-2500 mg/dL. The side

effects that occur are dyspepsia, diarrhea and lactate

The Effectiveness of Pare Extracts (Momordica Charantia L) in Lowering the Level of Blood Glucose on Wistar Rat (Rattus Norvegicus)

with Hyperglycemia

accidosis. Therefore, to reduce complications, the

initial oral dose of metformin given is 1500 mg

(Suherman, 2009).

The Determination of Glucose Volume Given to

the Samples

The volume of glucose given was based on the VAO

recommendation. A 10% glucose solution was given

after the initial blood glucose measurement was

performed. 2 ml of glucose solution was orally given

once. Previous research by Sari proved that the 2 ml

of 10% glucose solution can increase blood glucose

levels. The peak glucose level will occur for 30

minutes to 1 hour and return to normal within 2

hours.

The Treatment of Sample Animals

The white-mouse treatment group was divided into 5

groups. The first group of mice were given 10%

glucose and aquadest. The second group of mice

were given 10% glucose, and comparative drugs

(metformin). The third group of mice were given

10% glucose, and the pare extract at a dose of 100

mg/kgBW. The fourth group of mice were given

10% glucose and the extract at a dose of 200

mg/kgBW. The fifth group of mice were given 10%

glucose and the extract at a dose of 400 mg/kgBB

referring to the dose range in the previous study by

Adewole

. The post test were conducted done 4

times, that is at minute 65, minute 95, minute 125

and minute 155.

Data Analysis

The data of how blood glucose works was being

tested for its normality and homogeneity, to

determine whehther the data is normally distributed

or not. The normally-distributed data was then

proceeded using the Anova test with Duncan's

multiple-range test.

Research Ethics

The ethics of this study were based on the highest

respect to the sample animals as one of God's

creatures. Referring to the research ethics, the

research guarantees was submitted to the Ethics

Commission of Faculty of Medicine Unsyiah to

obtain the etchical clearance prior to the research.

The ethical clearance of this research is written in

letter Number: 42/KE/FK/2017.

3 RESULT AND DISCUSSION

The data on the average blood glucose level was

measured and the results can be seen in the table 1

and figures

Table 1 The Average Blood Glucose Level of the Sample Animals

Legend:

P0 : negative control

P1 : Metformin

P2 : extract 100 mg/kgBB

P3 : extract 200 mg/kgBB)

P4 : extract 400 mg/kgBB)

Notes :

1. The value of blood glucose levels in units of mg/dl

2. Different Superscripts shows significant differences

Group Minutes

0 30 65 95 125 155

P0 85,2

172,5

159,7

158.5

159,4

143,7

P1 86,2

196,8

151

117,6

98,6

83,6

P2 82,4

174,3

147,3

135,5

119,1

110,5

P3 89,2

176,6

126,4

117,4

105,8

95,1

P4 85,5

182,5

150

115,3

103,6

85,5

SKIC-MHS 2018 - The 2nd Syiah Kuala International Conference on Medicine and Health Sciences

Figure 1 Graph of average decrease in blood

glucose level of the sample animals

In this study, the blood glucose level of the fasting

mice ranged between 82.4-86.2 mg/dl. This shows

that all mice had normal fasting blood glucose

levels. After 10% glucose was inducted, there was a

125% increase in mice’s blood glucose level. This

situation indicates that the induction of glucose had

successfully made all mice experiencing the

hyperglycemic conditions. Proven that all mice have

hyperglycemic condition, the intervention was done

by giving metformin, pare extract with dose 100

mg/kgBB, 200 mg/kgBB, and 400 mg/kgBB.

In the first 30 minutes after the intervention by

pare extract at minute 65, a significant decrease was

found in blood glucose levels in the group with

200mg/KgBB of pare extract. This was a significant

reduction compared to not only the negative control

group, but also with the standard treatment group,

i.e. metformin. This suggests that among all the

doses tested, the dose of 200mg/kg (P = 0.016) has

the most rapid effect in lowering blood glucose

levels, as compared with metformin.

After 60 minutes intervention of pare extract at

minute 95, there was a significant difference of

blood glucose level in metformin group, 200

mg/KgBB and 400mg/KgBB of pare extract

compared to the negative control group. However,

there was no significant difference among the three

groups, which indicates that the three groups had the

same capacity to lower blood glucose levels.

In the 125

minute, there was a significant

difference between P2 (100mg/kgBW) and

metformin (p = 0.009), but the potential was not as

big as metformin P3 (200mg / kgBB) and P4

(400mg / kgBW) (p = 0.000 ).

In the 155

minute, blood glucose levels

decreased in all treatment groups and metformin was

the lowest, but the difference was not significant

with P3 (200mg/kgBB) and P4 (400mg/kgBW), so it

was considered the same.

The Duncan test results also showed the average

decreased of blood glucose levels, the negative

control group was the smallest and not statistically

significant. This is because the aquadest given to this

group have no effect on the decrease of blood

glucose. Changes in blood glucose levels that occur

in the negative control group tended to be result of

the body's physiological response to the increased

glucose intake. After the glucose was given, the

glucose levels will increase initially but will return

to normal within 2 hours (Price, 2006).

Momordica charantia L extract given to the

sample animals decrease their blood glucose level,

i.e. male mice (Rattus norvegicus) induced by

glucose compared to the negative control. The

decrease in blood glucose levels in sample animals

was due to the active compounds that contribute to

decreased blood glucose levels in pare extract, they

are alkaloids, saponins, flavonoids and charantin.

Charantin works by activating AMP-activated

protein kinase (AMPK) which increases the

synthesis of glycogen and also increases the glucose

uptake in liver and muscle cells (Bagchi, 2012).

Alkaloids lowers blood glucose by inhibiting the

glucose absorption in the intestine, increasing

glucose transport in blood, stimulating glycogen

synthesis and inhibiting glucose synthesis by

inhibiting glucose 6-phosphatase enzyme, 1.6-

bisphosphatase fructose, and increasing glucose

oxidation through 6-phosphate glucose. The 6-

phosphatase glucose and 1,6-biphosphatase fructose

are enzymes that play a role in gluconeogenesis.

Inhibition of these two will decrease glucose

formation from substrates other than carbohydrates

(Tachibana, 2001).

Saponins works in the same way as the class of

α-glucosidase enzyme inhibitors to prevent the

absorption of glucose, so that the amount of

monosaccharides absorbed by the intestine

decreases. Saponin compounds that act as

antihyperglycemia is triterpene saponins, which

prevents the glucose transport to the small intestine

which is a place for glucose absorption; therefore

preventing the rise of glucose level in blood

(Mohammady Elatar, 2012).

Another compound in Momordica charantia L

that plays a role in lowering the blood glucose level

is flavonoids. Flavonoids have the hypoglycemic

effects with several mechanisms, i.e. by inhibiting

the glucose absorption, increasing glucose tolerance,

acting like insulin, increasing the glucose uptake by

peripheral tissues and regulating enzymes that play a

role in carbohydrate metabolism (Brachmachari,

2011).

From the above description, it can be seen that

Momordica charantia L has active substances that

potentially act as anti-hyperglycemia. However, in

this study we did not isolate each of the active

The Effectiveness of Pare Extracts (Momordica Charantia L) in Lowering the Level of Blood Glucose on Wistar Rat (Rattus Norvegicus)

with Hyperglycemia

ingredients.

Concerning the effective dose in lowering blood

glucose levels, it was found that the doses of P3

(200mg/kgBB) and P4 (400mg/kgBW) can lower

blood glucose levels inthe same way of certain anti-

hyperglycemic, i.e. metformin. In addition, the

200mg/kgBB extract has the most rapid onset of

action compared to other doses, including

metformin. The 200mg/kgBB can lower blood

glucose levels at minute 65, while metformin at 95

minutes. It was noted that at minute 65, the blood

glucose level was significantly decreased in

hyperglycemic mice. At the same time, however,

there were no significant decrease in the blood

glucose levels of the other groups.

4 CONCLUSION

Based on the research results and discussion above,

it can be concluded that the ethanol extract of pare

(Momordica charantia L) effectively decrease the

blood glucose level of male Wistar mice (Rattus

norvegicus) induced by glucose. The most effective

dose for the pare ethanol extract (Momordica

charantia L) to lower the blood glucose level of

Wistar (Rattus norvegicus) male mice induced by

glucose was 200 mg/kgBW.

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SKIC-MHS 2018 - The 2nd Syiah Kuala International Conference on Medicine and Health Sciences