Effect of Ethanol Extract of Celery (Apium graveolens L) against

Urea and Creatinine Level in Male Wistar Rats on Ethylene Glycol

Induced Nephrolithiasis

Marline Nainggolan

, Kasmirul Ramlan Sinaga

, Siti Morin Sinaga

, Sony Eka

Nugraha

Department of Pharmaceutial Biology,

Faculty of Pharmacy, Universitas Sumatera Utara, Medan, 20155 Indonesia.

Department of Pharmaceutical Technology,

Faculty of Pharmacy, Universitas Sumatera Utara, Medan, 20155, Indonesia.

Department of Pharmaceutical Chemistry,

Faculty of Pharmacy, Universitas Sumatera Utara, Medan, 20155, Indonesia.

Keywords: Nephrolithiasis, Ethylene Glycol, Celery, Urea, Creatinine

Abstract: The use of celery in Indonesia was known as cooking spice to enrich the taste of food, has not been widely

used as herbal medicine either in the form of juice or extract. The Pre-clinical testing of celery herbs in addition

to diuretics it is also to treat sore eyes, sprains, rheumatism, hypertension, and as a hair tonic. The celery Herb

is rich in the content of flavonoids and potassium that can be used to dilute kidney stones. The aim of this

study was to investigate effect of ethanol extract of celery (Apium graveolens L) against urea and creatinine

level in male wistar rats on ethylene glycol induced nephrolithiasis. In vivo test was performed male wistar

which divided into 6 groups. Group I as normal control without any tratment, Group II as negative control

was induced ethylene glycol, Group III as positive control was given Batugin Elixir, Group IV, V and VI were

given ethanol extract of celery a dose of 50, 100 and 150 mg/kgBW respectively. Blood was collected by

cardiac puncture then measured level of urea and creatinine. The data were analyzed statistically using

ANOVA (analysis of variance). The results showed that ethanol extract of celery has an effective activity at

a dose of 150 mg / kgBW (P < 0.05) to negative group in decreasing level of urea and creatinine male wistar

rats on ethylene glycol induced nephrolithiasis and it showed a dose-dependent manner.

1 INTRODUCTION

Urolithiasis is a formation of crystal stone that formed

in the kidney and known as nephrolithiasis, its stone

also found in the urinary tract, bladder and ureters

(Selvam et al., 2001). The prevalence of urolithiasis

>10% with expected recurrence rate of >50%. The

incidence of nephrolithiasis in worldwide is high, and

around 80% of nephrolithiasis cases are found as form

of calcium oxalate or mixed with stone of calcium

phosphate (Brikowski et al., 2008).

Epidemiological

studies reported that the cases of nephrolithiasis

disease are higher between the ages of 20–40 in both

in men and women, men is more prevalent as 12%

than in women as 6% (Lopez et al, 2010).

Various study regarding to nephrolithiasis disease

has been isvestigated, such as radiological treatment

and laboratory determination. Both are not efficiently

to determine the mechanisms and causes of stone

deposite in the kidney and urinary tract (Tiselius et

al., 1980). However, some factors that would be

responsible induced kidney stone formation have

been studied recently (Curhan et al, 1997). It is

reported that the urine is saturated with material

compund which is a insoluble substance, it is leads to

formation of crystals and aggregate into stone form

(Sakhaee et al, 1993).

The nephrolithiasis needs

curative and preventive treatment. Nowdays, there are

no good modern medicine, that can dissolve stone

form in the part of kidney or urinary tract, therefore

the physicians is considering to alternative medicine

or treatment to heal the nephrolithiasis disease (Coe

at al, 1992).

The other medical treatment of

nephrolithiasis was surgical removal treatment of the

stones (Uribarri et al, 1989). Recently treatment

including extracorporeal shock wave lithotripsy

(ESWL) adn percutaneous nephrolithotomy (PCNL)

are not guarantee the recurrence of the disease

(Srisubat et al., 2009). Moreover, it can cause various

side effects such as bleeding, increasing blood

742

Nainggolan, M., Sinaga, K., Sinaga, S. and Nugraha, S.

Effect of Ethanol Extract of Celery (Apium graveolens L) against Urea and Creatinine Level in Male Wistar Rats on Ethylene Glycol Induced Nephrolithiasis.

DOI: 10.5220/0010087107420746

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

742-746

ISBN: 978-989-758-449-7

pressure and also kidney fibrosis (Wadhwa et al,

2007).

In the traditional cure, most of the sources

were taken from plants which has been proved

empirically in community that it is believe to heal

many disease. Herbal medicines are known have

lower side effects and efficacious than modern

medicine and it also reduce the prevalent of the

recurrece cases. Phytochemical properties such as

flavonoids spread widely in plants, and its biological

effect such as antioxidant have been reported in

various studies such as,

cancer, coronary heart

diseases and

diabetes (Hopper et al., 2008).

Herbal

Medicine have lower side effects compare to

chemical modern drugs because its antioxidant

activity has a role in lowering the toxicity of drugs

(Bent et al, 2004).

Today herbal medicine are used as

an alternative treatment to chemical drugs including

in treating nephrolithiasis (Butterweck and Khan,

2009).

Recent studies reported that the high content

of flavonoids plant could effectively inhibit the

formation of kidney stone in form of CaOx stones in

vivo and in vitro, correlating with its biological

activity such as anti-inflammatory, diuretic,

antioxidant, antibacterial properties and other

protective activity (Rad et al, 2011). Based on the

explanation, it showed that the flavonoids or

flavonoid-rich plant extracts has potential activitiy as

anti-nephrolithiasis.

Celery (Apium graveolens L) is a apiaceae family

plant, and grow in Europe and the tropical and sub

tropical regions of Africa and Asia. Beneficial

compounds are found in celery such as flavonoids

vitamins A and C, frocoumarin glycosides, limonene

and, selinene (Sowbhagya, 2014). Flavonoids content

of celery are the reason that celery has potential as

anti-urolithiasis activities

2 MATERIAL AND METHODS

2.1 Chemical

Ethanol 96%, aqudest, ethylene glycol, ammonium

chloride, Creatinine kit, Ureum kit

2.2 Plant Collection

Celery herb (Apium graveolens L) was collected from

local market at Padang Bulan, Northern Sumatra,

Indonesia. The plant samples authenticated by

Indonesian Institute of Science, Research Center of

Biology, Bogor, Indonesia.

2.3 Extraction of Celery Herb

An amount of 500 g dried Celery herb were crushed

in a blender, then macerated in ethanol 80 % for 5

days thereafter countinue to remecerated for 2 days.

The solvent was evaporated at low pressure with a

temperature of not more than 40 ° C using a Rotary

evaporator, then dried using freeze dryer (Nugraha et

al, 2018).

2.4 Phytochemical Screening of

Ethanol Extract Celery Herb

Phytochemical screening carried out on ethanol

extract celery herb includes examining the chemical

secondary metabolites of alkaloids, flavonoids,

glycosides, tannins, saponins, triterpenoids, and

steroids

2.5 Animals and Blood Sample

An amount of 30 male wistar rats used in this study,

weighing around of 180-220 g. The blood sample

was collected by cardiac puncture.

2.6 Experimental Design

The animals were given solution of 0.75% ethylene

glycol and also solution of 2% ammonium chloride

as much as 1% body weight orally to induce

Nephrolithiasis for 14 days.

Group I : as nornal control

Group II : as negative control (Na-CMC 0,5 %)

Group III : as positive control. Rats were received

Batugin Elixir 0.5 ml/ 200 g BW

Group IV : Rats were received ethanol extract of

celery herb at a dose of 50 mg/kg BW

Group V : Rats were received ethanol extract of

celery herb celery at a dose of 100 mg/kg BW

Group VI : Rats were received ethanol extract of

celery herb at a dose of 150 mg/kg BW

Treatment was given orally after ethylene

glycol induced. Treatment were given as long as 10

days, than in the day of 11, the blood was collected

from cardiac to determine the level of urea and

creatinine. Determination level of urea and creatinine

were measured using “Cobas Integra”.

2.7 Statistical Data Analysis

Data were analyzed using ANOVA and continue to

Tukey’s Multiple Comparison Test. Significance P

values were set at 0.05. Values for all measurements

are expressed as the mean ± SD

Effect of Ethanol Extract of Celery (Apium graveolens L) against Urea and Creatinine Level in Male Wistar Rats on Ethylene Glycol

Induced Nephrolithiasis

743

3 RESULT

3.1 Phytochemical Screening

Qualitative analysis of the various phytochemical

properties in the celery herb were done by qualitative

determination methods. the celery herb contains of

glycosides, alkaloids, steroids, tannins and flavonoids

compounds.

3.2 Level of Urea and Creatinine

Level of urea and creatinine was illustrated the

effectiveness of ethanol extract of celery herb to

inhibit nephrolithiasis that caused by ethylen glycol

induction. The level of urea and creatinine showed in

table 1 and table 2. Some study showed that ethylene

glycol induced the formation of kidney stone form in

albino Wistar rat. The process of supersaturation in

the urine which is contact to various forming

compound is the major factors in the kidney stone

deposition (Brikowski et al., 2008).

Using ethylene

glycol as an inducer of kidney stone formation is the

most suitable method and it is similar to kidney stone

that found in humans which cause the kidney damage

(Brikowski et al., 2008). Degradation of renal

function characterized by elevated levels of urea and

creatinine (Meier et al, 2004). In the present study, we

investigated the effect of celery (Apium graveolens L)

ethanol extract against urea and creatinine level in

male wistar rats on ethylene glycol induced

nephrolithiasis.

Urea is the final product of protein metabolism

and amino acids containing nitrogen One of the

important roles of the kidneys is to eliminate these

potentially toxic substances from the body. If there is

a decrease in kidney function, the blood urea nitrogen

level (BUN) increases ( Meier et al., 2004). BUN

measurements was one of kidney health marker.

Measurement of blood serum urea level of rats was

done to determine the effect of induction of ethylene

glycol 0.75% and 2% ammonium chloride to kidney

damage, and the effect of variation dose ethanol

extract celery herb on blood serum urea level of rat.



Table 1: Level of urea

No Group Level of urea Mean + SD

1 Normal control 40 41 41 42 39 40.6 + 1.1*

2 Negative control 57 56 58 55 56 56.4 + 1.1

3 Positive control 43 44 42 42 43 42.8 + 0.8*

4 Extract 50 mg/kgbw 47 48 50 49 47 48.2 + 1.3*

5 Extract 100 mg/kgbw 47 45 45 46 44 45.4 + 1.1 *

6 Extract 150 mg/kgbw 39 42 43 41 37 40.4 + 2.4 *

Where

* (Significantly different to negative control) P<0.05

# (Significantly different to positive control) P<0.05

Table 1: Level of creatinine

No Group Level of creatinine Mean + SD

1 Normal control 0.26 0.27 0.27 0.25 0.24 0.25 + 0.013 *

2 Negative control 0.45 0.49 0.48 0.45 0.48 0.47 + 0.018

3 Positive control 0.24 0.23 0.24 0.25 0.24 0.24 + 0.07*

4 Extract 50 mg/kgbw 0.37 0.44 0.45 0.42 0.45 0.42 + 0.033

5 Extract 100 mg/kgbw 0.37 0.40 0.39 0.35 0.37 0.37 + 0.019 *

6 Extract 150 mg/kgbw 0.33 0.30 0.28 0.29 0.30 0.30+ 0.018 *

Where

* (Significantly different to negative control) P<0.05

# (Significantly different to positive control) P<0.05

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

744

Creatinine was a metabolism molecule product

which is produced from the muscle metabolism.

Creatinine was an important molecule in the process

of energy production in the muscle, it is produced

from creatine. Every day, about 2% of the creatine

in the body, it was converted into creatinine form

(Horio, 2014). This molecules are transported into

the kidneys through blood vessel. The kidneys filter

out and remove of the creatinine in the urine. The

creatinine was a reliable indicator to determine the

function of.kidney.

The results of rat blood serum testing in each

group were statistically analyzed by spss to compare

the differences between treatment groups to negative

control and positive control. Based on Table 1, it

showed that the effective dose of ethanol extract

celery herb was 150 mg/kg BW and it showed dose

dependent manner. Level of urea in treatment group

of ethanol extract 150 mg/kgBW was 40.6 showed

there was no different to positive control (P>0.05)

and significant different to negative control (P<0.05).

Table 2 showed that effective dose of ethanol

extract celery herb to creatinine level of rat induced

ethylen glycol was 150 mg/kgBW with level of

creatinine was 0.30. Level of creatinine was not

different to positive control (P>0.05) and significant

different to negative control (P<0.05). Content of

flavonoids in ethanol extract celery herb prevents

oxidative stress in the kidney by increasing

glutathione s-transferase (GSH) antioxidant activity ,

increasing GSH synthesis and trapping directly ROS

formed by donating H atoms to free radicals resulting

in non-reactive free radical compounds and non-

reactive radical flavonol compounds that can

improve kidney function (Kang et al, 2016). In this

research, elevation of urea and creatinine were

decreased by treatment with ethanol extract celery

herb and thereby improved kidney functions in

nephrolithiatic condition.

4 CONCLUSION

Ethanol extract of celery herb at the dose of 50,100

and 150 mg/kgbw have effect in decreasing level of

urea and creatinine. Treatment dose which has

effective activity was 150 mg/kgbw and it showed

dose dependent manner.

ACKNOWLEDGEMENT

Research funded by Universitas Sumatera Utara in

accordance to the contract of research

implementation TALENTA Universitas Sumatera

Utara funding year 2018 No. 2590/UN5.1.R/PPM/

2018, date : 16 March 2018.

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