Histopathology of Rat Kidney Organ Due to Ethanol Extract of

Phaleria Macrocarpa Treatment Induced by Isoniazid

Ali Napiah Nasution* and Sri Lestari R. Nasution

Faculty of Medical, Universitas Prima Indonesia, Indonesia

Keywords: Phaleria macrocarpa, INH, Histopatology of the kidney.

Abstract: The use of anti-tuberculosis drugs has side effects on human organs, especially the kidneys. Phaleria

macrocarpa is now widely used as traditional medicine. The content of phaleria macrocarpa contains high

antioxidants and this fruit is spread in North Sumatra. This study aims to determine the histopathological

picture of induced kidney organ isoniazid (INH) by giving ethanol extract of phaleria macrocarpa. Animals

test used in this study were 9 wistar rats which were divided into 3 treatment and experiments groups for 10

days. The results showed that the histopathology of kidney organ caused by INH were highly damage.

However, giving Ethanol Extract of Phaleria macrocarpa at a dose of 3.4 g / 200 g was able to protect the

kidneys (nephroprotector) from the toxicity of isoniazid drugs with minimal damage or almost normal. The

administration of phaleria macrocarpa extract together with the anti-tuberculosis drug INH shows the

protective effect of the kidneys against the INH drug.

1 INTRODUCTION

In 2030 the World Health Organization (WHO)

devised a strategy to end tuberculosis (WHO, 2019).

Indonesia also aims to become a tuberculosis-free

country by 2050 (MOH RI, 2018). An adherence to

taking anti-tuberculosis drugs plays an important role

in the success of the treatment of tuberculosis

(Sitorus, Fatmawati and Rahmaniah, 2017).

Development of rapid industrialization generated

many negative consequence for environmental

pollution affecting human health. The previous

research of Nasution et all., (2015) explained that

P.macrocarpa contained hydrogen stretching due to

inter and intramolecular interaction of alcohol, phenol

and carboxylic acid. This functional group will

contribute to preventing metal ion from destroying

organs.

The kidney is the second most common organ

after the livertarget destroyer by xenobiotics. This

isdue to many chemical substancesexcreted in the

urine. One of the most common parts of the

kidneydamage caused by chemicals is the proximal

tubule. Some compounds that can be xenobiotic

include alcohol / ethanol andgentamicin (Panjaitan,

2003).Microscopic changes onkidney including

changes in the structure of the glomerulus, swelling

or enlargementkidneys and increased numbers of fat,

protein and water cells. This effect willchange the

ability of the kidneys to function normally (Booggan,

2003).

The most common side effect in patients taking

first-line OAT is nephrotoxicity (Pratiwi, 2018).

Histologically the administration of isoniazid causes

mild to moderate kidney damage (Muzika et al.,

2016).

Phaleria macrocarpa fruit is one of the ingredients

of traditional medicine that oftenly used to treat

various diseases and has antioxidant, antimicrobial,

and anti-inflammatory activity (Hendra et al., 2011).

Ramadhan Research, 2019 also stated that the

phaleria pacrocarpa fruit has an effect as a

nephroprotector in mice which induced by

paracetamol.

The results of Parapaga, Durry and Lintong, 2018

stated that antioxidant activity can inhibit and prevent

oxidative damage. Antioxidants can also reduce the

production of free radicals and protect body cells

from oxidative stress (Octaria, 2019).

According to Yatman (2012), the antioxidant

process throughoxidation and reduction reactions that

form oxidizing free radicalswith reactive oxygen.

Because of its reactivity, free radicals willoxidizes

beneficial substances for the body causing a

numberdamaged body tissue. Hence it is easily

Nasution, A. and R. Nasution, S.

Histopathology of Rat Kidney Organ Due to Ethanol Extract of Phaleria Macrocarpa Treatment Induced by Isoniazid.

DOI: 10.5220/0010296202430247

In Proceedings of the International Conference on Health Informatics, Medical, Biological Engineering, and Pharmaceutical (HIMBEP 2020), pages 243-247

ISBN: 978-989-758-500-5

243

oxidized, free radicals, for that matterperoxyl radical

(ROO) will oxidize xanthone rapidly so that it is

radicalthe peroxyl will turn into R-H. This change

occurs because of the oxygen moleculereduced by

garsinone B as a xanthone derivative, the reaction is

inhibitoryfree radicals of various types.Free radicals

can interfere with cellular function by

performinglipid peroxidation resulting in damage to

cell membranes. This damage can becauses changes

in electric charge in cells, changes in osmotic

pressure,causes cell swelling and ends in cell death.

Nakatni et al., (2004) conducted a study on the in

vitro anti-inflammatory activity of γ-mangostin

against the synthesis of PGE2 and cyclooxygenase

(COX) in glioma cells mouse C6. These two

compounds and enzymes are the most important

mediators in the inflammatory reaction. From this

research results can be made:mangostindirect PGE2

production in the inflammatory process.

In order to develop research on extracts of the

crown of the godsas a herbal medicine, this research

was conducted with the aim to find outthe effect of

giving the extract of the Dewa's Crown on the

histopathological picture of the kidney of rats which

was induced by isoniazid and the optimal dose of the

extract had an effect on the liver and kidneys. The

results obtained are expectedused as a source of

information about the benefits of extract of the crown

of gods especiallyon the kidneys and serve as the

basis for further researchdevelopment of mangosteen

peel extract as a standardized herbal medicine.

This study aims to determine the

histopathological picture of kidney organ induced by

INH (Isoniazid) by giving ethanol extract of Phaleria

macrocarpa fruit.

Nasution et al., (2019) Cd(II) ion is a heavy metal

that has atoxic abilty in the human body. phaleria

macrocarpa has been used as anticancer, Diabetes

Mellitus and antimicrobe because its consists of

flavonoid, steroid and tannin. The result of

exsperimental rats exposed witg Cd(II) ion, there are

significant decreasing of all the observed parameters

including MAD, SGOT and SGPT with percentage

71,5 %, 72,1 % and 93, 6 % respectively. The rats

given with the antidote of phaleria macrocarpa flesh

fruit were able to protect the liver from damage due

to exposure to Cd(II) as seen from the decrease in

liver function enzyme parameters namely SGOT and

SGPT

2 METHOD

This research (conducted at the Pharmacy

Laboratory, University of North Sumatra in

December 2019) used an experimental design with a

post-test only control group design pattern.

Experimental samples were 9 male Wistar white rats

(Rattus Novergicus), weight 150-200 grams were

used. Rats were in accordance with the code of

conduct issued by the Ethics Committee of the

Faculty of Medicine, University of Prima Indonesia.

The tools used were laboratory glass, analytical

balance, blender, incubator, oven, rotary evaporator,

water bath, animal scales, animal house, injection

equipment, surgical instruments, microtome, object

glass, cover glass, light microscope. The ingredients

used were phaleria macrocarpa, aquadest, isoniazid,

CMC-Na 0.5%, 10% formalin buffer, 96% ethanol,

preparations dyes.

The phaleria macrocarpa fruit was separated from

the skin and seeds and washed with running water.

The phaleria macrocarpa meat was cut into small

pieces and then dried and blended until smooth into

powder, then put into a container that can be tightly

closed.

The 500 mg simplicia powder was macerated with

96% ethanol, then evaporated with a rotary

evaporator at ≤70oC. Re-evaporated with waterbath

(water bath) until the extract results become thick

(Lestari, 2019; Ramadan, 2019).

Nephroprotector Effectiveness Test Samples were

divided into 3 groups, so that each group amounted to

3 rats. The treatment of each group was as follows:

Control group (P1) without treatment (only aquades).

Treatment Group 2 (P2) isoniazid 300 mg / 70 kg BW

induced. Treatment group 3 (P3) was given ethanol

extract of Crown Fruit of a dose of 3.4 g / 200 g BW

with isoniazid induction dose of 300 mg / 70 kg BW.

The treatments were orally contucted once per day for

10 days. At the end of the experiment, the rats were

anesthetized with chloroform before being dissected,

kidney organs were taken and then put into containers

containing 10% formalin buffer that had been labeled.

Preparations made with a thickness of 4-6 mm,

stained with HE and viewed under a microscope to

see histopathological changes.

3 RESULTS

3.1 Histopathology of the Kidney

Organs

The results of histopathology of the renal organs

examined in the control group are given in Figure 1.

In the control rat the glomerulus and renal tubules

were seen in normal condition, the nucleus was

clearly visible and the renal capsule was also clearly

HIMBEP 2020 - International Conference on Health Informatics, Medical, Biological Engineering, and Pharmaceutical

244

visible. Mice induced with isoniazid 300 mg / 70 kg

bw (KP2) were found with moderate glomerular

atrophy with necrosis. We found damage in the

kidneys as shown in Figure 2.

Mice given Ethanol Extract of Phaleria

Macrocarpa (EEPM) with isoniazid induction (KP3)

were seen with mild glomerular atrophy with

necrosis. In contrast to those who were only given

INH only in group 2, there was a lot of damage and

necrosis in the kidney organs. The administration of

the extract phaleria macrocarpa fruit can proverly

protect the kidneys from isoniazid induction.

Figure 1. Histopathology of rat kidney organ controls (P1).

10 x 10 magnification

Figure 2. Histopathology of rats kidney organ (P2) = INH

300 mg / 70 kg bw.

Description: glomerular atrophy (black arrow).

Figure 3. Histopathology of rats kidney organ (P3) =

EEPM + INH. Description: glomerular atrophy (black

arrow).

According to Zakhari et al., (2006), the results of

alcohol metabolism that occur inliver, namely the formation

of acetate, acetyldehyde, and increased reactive

oxygenspecies (ROS) enter the systemic blood circulation

which can damagethe structure of the cells of the extra

hepatic tissue namely the brain, lung, heart and

tissuekidney. ROS which are highly reactive can cause the

breakdown of molecular complexescellular (Wu and

Caderbaum, 2005). After ethanol intoxication,the balance

between prooxidants and antioxidants is disturbed

thuscauses oxidative stress from biomolecules, such as fat,

protein, or DNA,and ultimately cause cell damage (Das and

Vasudeven, 2007).

According to Yatman (2012), the process of

antioxidants through oxidation reactions andreduction

which forms free radicals which are oxidizing agents

withoxygenreactive. Because of their reactivity, these free

radicals willoxidize substances that arebeneficial to

thebody, causing a number of body tissues to be damaged.

Bybecause it is easily oxidized, free radicals, in this case

theperoxyl radical (ROO) willoxidizes xantones quickly, so

that the peroxyl radical will changeto be R-H. This

changeoccurs because the oxygen molecule is reduced by

garsinon Bas a xanthone derivative, its reaction can inhibit

free radicals of varioustype.

The molecular structure of each of the cell and tissue

flavonoid groupsthe body is constantly exposed to the

damaging effects caused by free radicalsand reactive

oxygen species (ROS) free radicalsnormally formed during

oxygen metabolism or induced by damageexogenous. Free

radicals can interfere with cellular function by

performinglipid peroxidation resulting in damage to cell

membranes. This damage can becauses changes in electric

charge in cells, changes in osmotic pressure,causes cell

swelling and ends in cell death. Flavonoidscan interfere

with more than 3 different free radical generating systems,

andcan also enhance endogenous antioxidant function.

Antioxidant activityhere is the antioxidant mechanism of

flavonoids, namely binding radicalsdirectly (direct radical

scanning venging), through nitric oxide, xanthinoxidase,

immobilization of leukocytes, interactions with other

enzyme systems (Nijveldtet al., 2001).

4 RESULTS

This research was conducted to determine the

effectiveness of etal extract of Phaleria macrocarpa

fruit on nephroprotectors in isoniazid-induced white

rats. Treated the rats with the EEPM at a dose of 3.4

g / 200 g bw can neutralize kidney damage due to

isoniazid with kidney histology indicators in mice.

Based on the results of research in the negative

control group (mice without treatment that was only

given aquadest) had a slight inflammation.

Supposedly in the negative control group there was

no damage to hepatocyte cells, this might be caused

by external variables that could not be controlled,

such as the psychological condition of the rat and the

Histopathology of Rat Kidney Organ Due to Ethanol Extract of Phaleria Macrocarpa Treatment Induced by Isoniazid

245

initial condition of the liver and kidney of the rat

before treatment. Changes in the environment of mice

also affect the behavior patterns of mice (Suharyadi,

Sukohar and Muhartono, 2014).

The histopathological picture assessed in the

kidney organ in the form of glomerular atrophy is

characterized by a decrease in the size of the

glomerular tissue due to the reduced number of cells.

The widening of the tubular lumen was due to the

presence of a protein cast or protein deposition on the

tubules.

According to Anggriani(2008), microscopic

picture of epithelial cells of the proximal tubuleswell

with granular cytoplasm due to shifting of

waterextracellular into cells. This shift occurs

because the toxin causeschanges in the surface

electric charge of tubular epithelial cells, active

transport of ions and acidsorganic, and the ultimate

concentrating ability of the kidneycauses the tubule to

be damaged, the flow decreases. Picture of swelling

of these cellscalled albuminous degeneration or

parenchymal or cloudy degenerationswelling (cloudy

swelling). This may cause the tubular lumenproximal

narrowing to close. According to researchconducted

by Liu et al., (2002) who stated that giving

ethanol50% with a dose of 10 ml / kg of rats can cause

kidney damage.

The treatments with the Phaleria macrocarpa

extract for 10 days did not cause any toxicity, because

there were no deaths and poisoning in white rats

during the study. Based on the histopathology that has

been studied, the EEPM is able to protect the kidneys

due to isoniazid induction because it has high

antioxidants.

The antioxidant content of flavonoids as

inhibitors of CYP3A activity and acts as a free radical

scavenger (Hassanin, Abd El-Kawi and Hashem,

2013). Flavonoid compounds contained in the

phaleria macrocarpa fruit have an influence in

protecting the kidneys.

5 CONCLUSIONS

The results showed that an effect after giving an

EEPM at a dose of 3.4 g / 200 g bw on the kidney

tissue damage were highly reduced as evidenced by

histopathological examination. Preventive use of the

EEPM can act as a protective kidney when

consuming OAT INH together.

6 SUGGESTION

The next researcher is suggested to design experiment

using the phaleria macrocarpa fruit in the different

dosage of concentration with a longer trial period.

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