Antihyperglycemic Activity of Ethanolic Herb Extract of Ceplukan

(Physalis angulata L.) in Diabetic Hypercholesterolemia in Male

Hamsters

Elly Wardani

, Dwitiyanti, Sediarso, Dwina Puspandiyah

Faculty of Pharmacy and Science, Universitas Muhammadiyah Prof. Dr. Hamka, Jakarta, Indonesia

Keywords: Physalis angulata L., Ceplukan herb extract, Antihyperglycemic, blood glucose

Abstract: Diabetes mellitus is a disease characterized by hyperglycemia as well as progressive changes to the

pancreatic beta cell structure. This study was conducted to determine the antihyperglycemic activity of

ceplukan herb extract (Physalis angulata L.) in alloxan-induced male Syrian hamsters and high cholesterol

feed. The study used 24 hamsters divided into six groups. Group I were given a standard diet and regular

drinking water, Group II were given metformin dose 61,66 mg/kg body weight (BW), Group III were

alloxan-induced and high cholesterol feed, Groups IV, V and VI were given extract dose 60, 120 and 240

mg/kg BW respectively. The animals were induced alloxan monohydrate as well as were given high

cholesterol feed during treatment. Blood sampling was performed on the 29th and 44th day using a clinical

spectrophotometer. The results show significant differences between treatment groups (α <0,05), followed

by Tukey test. In conclusion, the preparation of herbal extract test ceplukan at doses of 120 and 240 mg/kg

BW could reduce blood glucose levels by 50.84% and 43.41% which is equivalent to metformin dose of

61.66 mg/kg BW with a percentage of 53.12%.

1 INTRODUCTION

Diabetes mellitus (DM) is a group of metabolic

disorders characterized by hyperglycemia and

abnormalities in carbohydrate, fat, and protein,

resulting from insulin secretion abnormalities,

insulin work or both (Dipiro, 2014). Diabetes

mellitus is divided into two types, namely type I DM

(5%-10% of cases), which is caused by damaged

pancreatic β cells suffered by hereditary or genetic

and type II DM (90% of cases) caused by insulin

resistance due to increased lipolysis, free fatty acid

production or increased glucose production in the

liver (Dipiro, 2015).

DM in Indonesia is a serious threat to health

development, can cause blindnesss, kidney failure,

diabetic feet (gangrene) so it must be amputated,

heart disease and stroke (Depkes, 2013). The World

Health Organization (WHO) Global Status Report

on NCD in 2010 stated that 60% of the cause of

death of all ages in the world is due to non-

communicable diseases and DM is ranked 6th as the

cause of death. Approximately 1.3 million people

die from diabetes and 4% die before the age of 70

years. In 2030, it is estimated that DM will be

ranked 7th as the cause of death worldwide and that

Indonesia will have 21.3 million people with DM

(Depkes, 2013). This condition will make Indonesia

on the 4

position after the United States, China and

India as countries that have the largest diabetics with

the largest population in the world (Depkes, 2012).

One of the medicinal plants that can be used as

an antidiabetic agent is ceplukan (Physalis angulata

L.). In West Java, ceplukan has been used as

diabetes drug (Sutjiatmo et al. 2011). Chemical

compounds of this plant include alkaloids,

flavonoids, saponins, fisalin, sterols/terpenes and

citric acid (Depkes, 1995). Traditionally, this plant

has been used to treat boils, and diabetes (Depkes,

1995).

Previous research has shown that the ethanol

extract of ceplukan leaves at 100 mg/kg BW can

reduce blood glucose levels in mice with a

percentage of 56.34% (Kasali, 2016). In isolation

research from ethanol extract of ceplukan fruits at 25

mg/kg BW and 50 mg/kg BW for 15 days can

decrease rat blood glucose level to 38,75% and

27,55% respectively (Raju, 2015). Another study has

Wardani, E., Dwitiyanti, ., Sediarso, . and Puspandiyah, D.

Antihyperglycemic Activity of Ethanolic Herb Extract of Ceplukan (Physalis angulata L.) in Diabetic Hypercholesterolemia in Male Hamsters.

DOI: 10.5220/0008240601330137

In Proceedings of the 1st Muhammadiyah International Conference on Health and Pharmaceutical Development (MICH-PhD 2018), pages 133-137

ISBN: 978-989-758-349-0

133

reported that the fraction of chloroform herb

ceplukan at a dose of 0,5 mg/20g BW, 1 mg/20g

BW and 2 mg/20g BW can reduce blood glucose

levels in mice and the dose 2 mg/20g BW can

reduce blood glucose levels of mice equally to

glibenclamid 0,013 mg/20g BW (Sunaryo et al.

2012).

2 MATERIALS AND METHODS

2.1 Materials

The ingredients used are ceplukan herb (Physalis

angulata L.). Determination was done at

“Herbarium Bogoriense” Botani section, Pusat

Penelitian Biologi - LIPI Cibinong.

Alloxan monohydrate from Sigma-Aldrich Co.,

USA. Metformin HCL from Sohan Healthcare Pvt

Ltd, Maharashtra. High cholesterol feed, glucose kit

reagent was using commercial kits purchased from

Human Diagnostics Worldwide Germany.

2.2 Animal Subjects

The experiment used twenty-four male Syrian

hamsters (Mesocricetus auratus), aged 3–4 months

and weighed around 80 g. The animals were housed

under standard environmental conditions. The study

protocol was approved by the Health Research

Ethics Committee of Universitas Muhammadiyah

Prof. Dr. Hamka, West Jakarta, Indonesian. The

reference number for approval was 02/18.05/005.

The animal subjects were acclimatized in a cage

for approximately one week to adapt to the new

environment. The animals were divided into 6

groups consisting of 4 hamsters.

2.3 Extraction

One kilogram of ceplukan dried powder (Research

Institute for Spices and Medicinal

Plants/BALITTRO, Bogor, Indonesia) was

macerated with 70% ethanol for 6 hours and was

stirred occasionally to reveal the active ingredients.

The mixture was allowed to stand for 18 hours. The

maceration result was then separated by filtration.

The reproduction process was repeated three times.

The maceration result was concentrated with a

vacuum rotary evaporator until it became a viscous

extract (Department of Health RI, 2008). The

concentrated result was put in a water bath at 40–

C for one day to obtain an extract with a constant

weight.

2.4 Phytochemical Screening

The phytochemical screening of ceplukan herb

extract included an examination of alkaloids using

Bourchardat reagents, testing flavonoids with

methanol, concentrated HCL and magnesium

powder. Saponin test were perfomed with foam

formation, tannin test with FeCl

1% reagent, and

Steroid testing with ethanol, concentrated H

were done if there is a red or purple color change

indicating the presence of triterpenoids and the green

color indicates steroid presence (Depkes, 1995).

Extract yield was determined by calculating the dry

extract weight obtained from the weight of dry

powder before extraction.

2.5 Category and Animals Subject

Treatment

The experiment was done with a complete

randomized design, using 24 white male, hamsters

divided into six groups consist of 4 rats.

a. Group I: The group was given a standard diet

and regular drinking water.

b. Group II: Positive Control, was given alloxan

monohydrate, high cholesterol feed and

comparative preparations.

c. Group III: Negative Control, was given alloxan

monohydrate, high cholesterol feed and Na-

CMC 0,5%.

d. Group IV: was given alloxan monohydrate, high

cholesterol feed and ethanolic herb extract of

ceplukan dose I.

e. Group V: was given alloxan monohydrate, high

cholesterol feed and ethanolic herb extract of

ceplukan dose II.

f. Group VI: was given alloxan monohydrate, high

cholesterol feed and ethanolic herb extract of

ceplukan dose III.

2.6 Method of Glucose Levels

Measurement

Blood sampling was taken on Day 29 and Day 44;

hamsters were first anaesthetized using ketamine

dose 10 mg/kg BW (Lacy et al. 2009). Blood

collection was performed at the eye orbital sinuses

after the animals were previously preoccupied for 12

hours. The blood was taken as much as one mL, and

was collected in a microtube. The blood was

centrifuged at 4000 rpm for 15 minutes to obtain the

serum (Suharmiati, 2003). Serum was taken ten μL,

was mixed with enzyme reagent (glucose reagent

kit) 1000 μL, then was homogenized using vortex

MICH-PhD 2018 - 1st Muhammadiyah International Conference on Health and Pharmaceutical Development

134

for 1 minute and was incubated for 5 minutes at

C blood glucose level were tested with a clinical

photometer (Human, 2012).

2.7 Data Analysis

The data using statistical analysis show that the

decrease in percentage data from the initial and late

glucose levels, baseline levels are levels after

alloxan induction and high cholesterol feed while the

final content levels after treatment. The data were

tested for normality and homogeneity and were then

analyzed with one-way ANOVA test with 95%

significance level (α = 0,05). If there is a significant

difference, the data were then analyzed with the

Tukey test (Santosa, 2010).

3 RESULTS AND DISCUSSION

Based on the results of the determination, indicating

that the plant is Physalis angulata L. included in

Solanaceae. Data extraction of ceplukan herb such

as weigh ceplukan fresh and dry, weigh of ceplukan

extrac and extract yield are given in Table 1.

Parts of ceplukan that used is the herbaceous

part. A report by The Department of Health (1995)

stated that chemical compounds found in ceplukan

include alkaloids, flavonoids, saponins, fisalin,

sterols/terpene and citric acid. Alkaloids, flavonoids,

saponins, and steroids could be obtained by

phytochemical screening. (Table 2). The results of

extract quality characteristics showed specific smell,

thick form and had a drying loss of 9.59% can be

seen in table 3.

Metformin was used as a comparative drug in

this study because it is a first-line antidiabetic drug

against DM2 patients with a history of obesity. In

this study, the test animals not only being treatment

to become hyperglycemia, but also induced to be

hypercholesterolemia. Then, induction of alloxan

will damage cells β Langerhans pancreatic cells to

inhibit the production of insulin. Therefore

metformin has a mechanism of increasing peripheral

tissue sensitivity of insulin (Suharmiati, 2003) and it

was chosen as a comparative drug preparation in this

study because the mechanism was aligned with the

alloxan to correct the damage caused by alloxan.

Feeding for the hypercholesterolemic hamsters

was performed for 44 days to maintain the stability

of cholesterol. On Day 22, alloxan was induced

intraperitoneally with a dose of 180 mg/kg BW.

Before blood sample collection, the hamsters were

fasted for 12 hours to avoid the effect of increased

glucose level after meal. Measurement of baseline

blood glucose level of the test animals were

performed on Day 29 to determine whether the

induced alloxan had successfully increased the blood

glucose level. After the level of glucose and

cholesterol were stated to increase pathologically

(the level of complication was achieved), then the

extract was orally administered by using feeding

tube. The extract was given every morning for 14

days. On Day 44, the final blood glucose level was

checked to see whether there is a decrease or not.

The data of the percentage of decrease of blood

glucose level was examined by normal distribution

test {(α = 0,202) > 0,05} and homogeneity test {(α =

0,104) > 0,05} which show that the data is normally

distributed and homogeneous. One-way ANOVA

test was run to test whether the mean of % decrease

in the blood glucose level is the same or

significantly different. The result {(α = 0,000) <

0,05} shows that there are significant differences

between all groups. Then the one-way ANOVA

results were further examined with the Tukey test to

Table 2: Results of Phytochemical Extract Screening

No. Chemical

com

oun

Extract

1. Alkaloi

2. Saponin +

3. Tanin -

4. Flavonoi

5. Steroi

6. Triter

enoi

ote: “+” (positive), “-“ (ne

ative)

Table 1: Data extraction results

No. Type Result

1. Ce

lukan Fresh 15 k

2. Ce

lukan Dr

1,5 k

3. Ce

lukan Powde

1,1 k

4. Macerate of Powde

1 kg

5. Extract 158,86 g

6. Extract Yiel

15,886 %

Table 3: Extract characteristics

No. Type of Test Extract

1. Organoleptic

Smell S

ecific

Colo

Brown Green

Form Thick, bitte

2. Flavors 9,59%

Antihyperglycemic Activity of Ethanolic Herb Extract of Ceplukan (Physalis angulata L.) in Diabetic Hypercholesterolemia in Male

Hamsters

135

know which group has, or has no significant

difference between normal control, negative control,

positive control, Dose 1, Dose 2 and Dose 3.

The-ability to decrease blood glucose is related

to the biological activity of compounds in ceplukan

plants. The compounds contained are flavonoids,

alkaloids, steroids, and saponins. The chemical

content of steroids can stimulate the release of

insulin from the pancreas to lower blood glucose

levels (Sediarso et al. 2008). Insulin will then work

to increase the transport of glucose from the blood

into cells by increasing the permeability of the cell

membrane to glucose. Once inside the cell, glucose

will then be used to generate energy. The liver and

muscles will convert glucose into glycogen and will

then be stored there. This conversion causes the

blood glucose level in the body decreases slowly

(Gustina, 2012).

Figure 1 shows the plasma glucose levels of the

experimental animals. The highest decrease in blood

glucose was shown by Dose 2 (120 mg/kg BW)

which was 50,84% compared to Dose 3 (240 mg/kg

BW) which was 43,41% and Dose 1 (60 mg/kg BW)

which was 34,74%. All pharmacologic responses

must have the maximum effect (Emax), regardless

of the high concentration that occurs, a point is

reached when adding the concentration does not

increase the response (Katzung et al. 2014). This

occurs in Dose 2 that was smaller than Dose 3, but

Dose 2 gave a greater decrease effect compared to

Dose 3, the maximum effect occurred in Dose 2;

although Dose 3 twice higher. Therefore, this study

shows that Dose 2 (120 mg/kg BW) has a better

activity in lowering blood glu

cose by 50,84%.

4 CONCLUSION

It can be concluded that ceplukan extract can lower

blood glucose levels in male hamsters with

hypercholesterolemia and hyperglycemia. Dose 2

(120 mg/kg BW) has a better activity in lowering

blood glucose by 50,84% and equivalent to

metformin dose 61,66 mg/kg BW which was

53,12%.

FUTURE RECOMMENDATION

The next stage of the research may focus on the

content of plant compounds and fractionation phase

as a decrease in blood glucose levels.

REFERENCE

Departemen Kesehatan RI. 1995. Farmakope Indonesia.

Edisi IV. Departemen Kesehatan Republik

Indonesia, Jakarta. Hal 7, 410

Departemen Kesehatan RI. 1995. Materia Medika

Indonesia. Jilid VI. Departemen Kesehatan RI.

Jakarta. Hal. 195,199

Departemen Kesehatan RI. 2008. Farmakope Herbal.

Edisi 1. Depkes RI. Jakarta

Depkes RI. 2012. Kemitraan Pemerintah Dan Swasta

Dalam Pengendalian Diabetes Melitus Di Indonesia.

http://www.depkes.go.id/article/view/2053/kemitraan-

pemerintah-dan-swasta-dalam-pengendalian-diabetes-

melitus-di-indonesia-.html. Diakses 8 Februari 201

Depkes RI. 2013. Diabetes Melitus Penyebab Kematian

Nomor 6 Di Dunia: Kemenkes Tawarkan Solusi

Cerdik Melalui Posbind.

http://www.depkes.go.id/article/view/2383/diabetes-

melitus-penyebab-kematian-nomor-6-di-dunia-

kemenkes-tawarkan-solusi-cerdik-melalui-

posbindu.html. Diakses 8Februari 2017

Dipiro JT, Talbert RL, Yee GC, Matzke GR, Wells BG,

Posey LM. 2014. Pharmacotherapy: A

Phatophysiologic Approach. Ninth edition. USA: The

McGraw-Hill Companies. Page. 2546

Dipiro JT, Talbert RL, Yee GC, Matzke GR, Wells BG,

Posey LM. 2015. Pharmacotherapy Hanbook. Ninth

Figure 1: The average percentage decrease in blood

glucose levels in the administration of ceplukan herb

extract

MICH-PhD 2018 - 1st Muhammadiyah International Conference on Health and Pharmaceutical Development

136

edition. USA: The McGraw-Hill Companies. Page.

161

Gustina NMR. 2012. Aktivitas Ekstrak, Fraksi Pelarut dan

Senyawa Flavonoid Daun Sukun (Artocarpus altilis)

Terhadap Enzim α-Glukosidasi sebagai Antidiabetes.

Dalam: Skripsi. Bogor. Institut Pertanian Bogor

Human. 2012. Glucose Liquicolor. Human Gesellschaft

for Biochemica and Diagnostica mbH. Germany

Kasali FM, Fokunang CN, Ngoupayo Joseph, Ngameni

Bathelemy, Fokunang ET, Njinkio Borgia, Kechia

Frederick, Kadima JN, Marthe Tsague, Victor Oyono,

Mbacham Wilfred, Ngadjui BT. 2016. Evaluation of

the Antidiabetic Properties of Hydro-Alcoholic Extract

and Its Fractions from Physalis peruviana L. Leaves

on Streptozotocin-Induced Diabetic Wistar Rats.

Journal of Diseases and Medicinal Plants Vol.2 No.6.

Cameroon

Katzung BG, Masters SB, Trevor AJ. 2014. Farmakologi

Dasar dan Klinik Ed 12. Penerbit Buku Kedokteran

EGC, Jakarta. Hal 54

Lacy CF, Abreg JA, Amstrong LL, Goldman MP, Lance

LL. 2009. E-book Drug Information Handbook. Edisi

17. Lexi-Comp for the American Pharmacists

Association

Raju Proka, Mamidala Estari. 2015. Anti-Diabetic

Activity Of Compound Isolated From Physalis

angulata Fruit Extracts In Alloxan Induced Diabetic

Rats. The American Journal Of Science And Medical

Research. Vol 1 No.1. India

Santosa S. 2010. Statistik Parametrik. Elex Media

Komputindo. Jakarta

Sediarso, Sunaryo H, Amalia N. 2008. Efek Antidiabetes

dan Identifikasi Senyawa Dominan dalam Fraksi

Kloroform Herba Ceplukan (Physalis angulata L.).

Jurnal Farmasi Indonesia.Vol. 4. Jakarta

Suharmiati. 2003. Pengujian Bioaktivitas Anti Diabetes

Mellitus Tumbuhan Obat. Majalah Cermin Dunia

Kedokteran No. 140. Surabaya. Hal. 8-13

Sunaryo H, Kusmardi, Trianingsih W. 2012. Uji Aktivitas

Antidiabetes Senyawa Aktif Dari Fraksi Kloroform

Herba Ceplukan (Physalis angulata L.) Terhadap

Penurunan Kadar Glukosa Darah dan Perbaikan Sel

Langerhans Pankreas Pada Mencit yang Diinduksi

Aloksan. Jurnal Farmasains. Vol 1 No. 5. Jakarta.

Hal. 251

Sutjiatmo AB, Sukandar EY, Ratnawati Yulia,

Kusmaningati Suswini, Wulandari Asri, Narvikasari

Suci. 2011. Efek Antidiabetes Herba Ceplukan

(Physalis angulata Linn.) Pada Mencit Diabetes

Dengan Induksi Aloksan: dalam Jurnal Farmasi

Indonesia Vol.5 no.4 Juli. Hal. 166-171

Antihyperglycemic Activity of Ethanolic Herb Extract of Ceplukan (Physalis angulata L.) in Diabetic Hypercholesterolemia in Male

Hamsters

137