The Effects of Aqueous Extract of Jamaican Cherry (Muntingia

calabura) on D-galactose-induced Liver Damage in

BALB/c Mice (Mus musculus)

Cempaka Jaga Pramudita

, Dwi Nur Ahsani

, Ika Fidianingsih

, Evy Sulistyoningrum

Faculty of Medicine, Universitas Islam Indonesia, Yogyakarta, Indonesia

Department of Histology, Faculty of Medicine, Universitas Islam Indonesia, Yogyakarta, Indonesia

Keywords: D-Galactose, Muntingia calabura, Hepatocyte Damage, Jamaican Cherry, Manja Roenigk, Aqueous Extract

Abstract: D-Galactose induces oxidative stress that will damage the hepatocytes. The aqueous extract of Muntingia

calabura leaves (AQMC) is known to have a potential antioxidant to prevent hepatocyte damage caused by

oxidative stress. This study aimed to determine the effects of AQMC leaf extract on liver damage induced by

D-Galactose in BALB/c Mus musculus. This was experimental research with a completely-randomized design

involving 20 hepatic samples (paraffin blocks and HE staining) from 5 groups. The groups included K1

(healthy), K2 (D-galactose-induced), K3 (D-galactose + AQMC 35 mg/kgBW), K4 (D-galactose + AQMC

75mg/kgBW), and K5 (D-galactose + vitamin C 28mg/KgBW). D-galactose was administered for 6 weeks

prior to the therapy (vitamin C or AQMC given for 4 weeks). Liver damage was observed in all fields of view

and described comprehensively. The degree of hepatocyte damage was calculated using the Manja Roenigk

scoring and analyzed using One-way ANOVA with post-hoc Tukey’s HSD test (CI = 95%, α = 0.05). AQMC

leaf extract could reduce D-galactose-induced liver damage in BALB/c Mus musculus. The hepatocyte

damage in the groups given AQMC therapy was less than that in the D-galactose negative control group (K1

= 73.5 + 2.39, K2 = 92.00 + 5.24, K3 = 69.00 + 2.79, K4 = 76.25 + 4.42, K5 = 77.25 + 6.48; p = 0.029).

AQMC at a dose of 35mg/KgBW showed more effective therapeutic potential against D-galactose (K3 =

0.003, K4 = 0.027; post-hoc toward K2). AQMC administration could reduce liver and hepatocyte damage of

BALB/c Mus musculus induced by D-Galactose at a potential dose of 35 mg.

INTRODUCTION

Aging can raise the levels of free radicals (ROS) in

the blood and tissues. Excessive ROS in an aging

process is induced by an imbalance between ROS and

antioxidant. Studies show that D-galactose induced in

animal models can describe the process of oxidative

stress in aging. D-galactose increases plasma

Malondialdehyde (MDA) levels (Sulistyoningrum et

al., 2019), raises MDA levels in the liver, and reduces

hepatic antioxidant (SOD) levels (Hadzi-Petrushev et

al., 2015). Induced D-galactose will also activate the

p-53 pathway, thus leading to cell apoptosis, and

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stimulate the p-21 pathway that plays a role in the cell

cycle (Bo-Htay et al., 2018).

Aging reduces the ability of the liver to regenerate

during cellular injury. Research shows that the

hepatocytes in aged rats lose the capability of entering

mitosis (Biondo-Simões et al., 2006), and the cells’

ability to recognize growth factors, such as EGF, also

decreases in aging rats (Schmucker & Sanchez,

2011). The reduced regenerative capability increases

the likelihood of cell damage and apoptosis. Cell

apoptosis in the liver is susceptible not only to

oxidative stress but also to genomic instability and

lipotoxicity (Zhong et al., 2017).

In addition to increasing cell apoptosis, the aging

process in the liver is also histologically marked by

202

Pramudita, C., Ahsani, D., Fidianingsih, I. and Sulistyoningrum, E.

The Effects of Aqueous Extract of Jamaican Cherry (Muntingia calabura) on D-galactose-induced Liver Damage in BALB/c Mice (Mus musculus).

DOI: 10.5220/0010490102020207

In Proceedings of the 1st Jenderal Soedirman International Medical Conference in conjunction with the 5th Annual Scientiﬁc Meeting (Temilnas) Consortium of Biomedical Science Indonesia

(JIMC 2020), pages 202-207

ISBN: 978-989-758-499-2

dilation of blood vessels, increasing number of

Kupffer cells, and escalation of cell degeneration

(Hashish, 2016). Therapy to reduce aging effects on

the liver should therefore be considered in order to

prevent age-related liver diseases, including hepatic

cirrhosis, liver cancer, or non-alcoholic fatty liver

disease (NAFLD)

Muntingia calabura (MC) is among the medicinal

plants with highly potential preventive properties

against oxidative-stress-induced cell damage. The

administration of methanol extract of Muntingia

calabura leaves (MMC) can prevent increased uric

acid levels and damage to the proximal renal tubules

in DM model rats (Safrida & Sabri, 2019). In

addition, MMC leaf extract given as premedication

for hepatotoxic substances (CCl4) has proved to

reduce the levels of parenchymal liver damage

(Zakaria et al., 2019). The potential protective role of

MC has also been observed in non-methanolic

extracts. A number of studies report the

administration of MC ethanol extract that can

suppress an inflammatory process and prevent gastric

ulcers in ethanol-induced rats (Aziz Ibrahim, 2012;

Lin et al., 2017; Sarimanah et al., 2017). Combined

MC-Ficus carica infusion is able to reduce SGOT and

SGPT levels in paracetamol-induced rats (Lalihatu &

Sudharmono, 2019). Furthermore, the administration

of MC as premedication can prevent carbonated-

drink-induced or ethanol-induced liver damage

(Murti et al., 2016).

2 MATERIAL AND METHODS

2.1 Research Design and Subjects

This research was purely experimental with a

completely-randomized design. The experiment was

conducted from April to October 2016 at the

Laboratory of Histology and Anatomical Pathology

of the Faculty of Medicine, Universitas Islam

Indonesia Yogyakarta after passing the ethical review

from the ethics committee of the Faculty of Medicine

of Universitas Islam Indonesia. This study used 20

livers of BALB/c mice (Mus musculus) obtained

from a previous study (Sulistyoningrum et al., 2019).

The inclusion criterion was the hepatic tissue

obtained from a previous research protocol. If

damage to the hepatic tissue was found

(microscopically or macroscopically) and thereby

resulting in difficulty to interpret the results, the tissue

was excluded. The 20 samples were obtained from 5

groups named K1 (healthy normal group), K2

(negative control group, D-galactose-induced), K3

(dose-1 treatment group, D-galactose-induced and

35mg/kgBW AQMC leaf extract), K4 (dose-2

treatment group, D-galactose-induced and

75mg/kgBW AQMC leaf extract), and K5 (positive

control group, D-galactose-induced and

28mg/KgBW vitamin C). D-galactose was

administered for 6 weeks while the treatment

(administration of AQMC or vitamin C) was given

daily for 4 weeks after induction. The hepatic tissue

was then transversely embedded in a paraffin block

and stained with HE staining.

2.2 Observation of Hepatic

Histomorphological Changes

The liver damage was observed descriptively and

semi-quantitatively. The changes observed in all

fields of view consisted of inflammation,

degeneration, necrosis, indistinctive cell boundaries,

and affected size of the central vein (Zulfi et al.,

2013). The levels of liver damage were calculated on

the basis of Manja Roenigk scoring on 50 cells (1:

normal, 2: inflammation, 3: degeneration, 4: necrosis)

in six identical fields of view for each sample (total

magnification of 400x). Inflammation was marked by

lymphocyte invasion, degeneration was manifested as

clear cytoplasm and giant cells in hepatocytes

(Sookoian et al., 2016), and necrosis was indicated by

changes in hepatocyte nuclei (pyknosis, karyorrhexis,

and karyolysis). The final score of liver damage in

each sample was obtained by multiplying the number

of cells by the Manja Roenigk scoring (Zulfi et al.,

2013). In addition, cell boundaries were classified

into distinctive, indistinctive, and fairly distinctive.

The size of central vein was then compared with that

of the healthy control group to obtain three categories

of size change named normal, slightly enlarged (>1.5-

2-fold), and enlarged (>3-fold).

2.3 Statistical Analysis

An analysis was performed on the semi-quantitative

data of liver damage scores. The normality was

examined using the Saphiro Wilk test while the

significance test involved One-way ANOVA

followed by Post-Hoc Tukey’s HSD test. All of the

statistical tests were done at a confidence level of

95% (α = 0.05)

3 RESULTS

The hepatic histomorphological changes during the

aging process (D-galactose-induced) include

The Effects of Aqueous Extract of Jamaican Cherry (Muntingia calabura) on D-galactose-induced Liver Damage in BALB/c Mice (Mus

musculus)

203

inflammation, degeneration, necrosis and a change in

the central vein size (Figure 1). The hepatic

histological changes were evident in the negative

control group (K2), and inflammation was found in

all of the study groups. The administration of AQMC

leaf extract improved the histopathological features

of D-galactose-induced liver damage in mice.

Meanwhile, the administration of vitamin C resulted

in histopathological features resembling those of

AQMC leaf extract at a dose of 75mg/kgBW (Table

1). The hepatocyte damage found in this study

showed a significant difference (p = 0.029, Table 2),

which was mainly found between the AQMC leaf

extract treatment group and the K2 group (dose of

35mg/KgBW: 0.003, dose of 75mg/KgBW: 0.027,

Table 3).

Table 1: Histological features of liver damage in different groups.

Inflammation Degeneration Necrosis Cell boundaries Central vein size

K1 + - - distinctive normal

K2 ++ +++ ++ indistinctive enlar

K3 + + + distinctive normal

K4 + ++ + fairly distinctive slightly enlarge

K5 ++ ++ + fairly distinctive slightly enlarge

Note: - normal, + mild, ++ moderate, +++ severe. K1: healthy normal group, K2: negative control group (D-galactose-induced), K3:

dose-1 treatment group (D-galactose-induced and 35 mg/kgBW AQMC leaf extract), K4: dose-2 treatment group (D-galactose-

induced and 75 mg/kgBW AQMC leaf extract), K5: positive control group (D-galactose-induced and 28 mg/kgBW vitamin C).

Figure 1. Hepatic histological features with HE staining and 40x objective magnification in all groups (in alphabetical order

for K1, K2, K3, K4, K5). Black arrow: normal liver, white arrow: inflammation, yellow arrow: degeneration, green arrow:

necrosis, VS: central vein.

JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )

204

Table 2: Mean Anova of hepatocyte damage in different

groups.

Group n Mean+SD p

K1 4 73.5+2.39 0.029

K2 4 92.00+5.24

K3 4 69.00+2.79

K4 4 76.25+4.42

K5 4 77.25+6.48

*: (p<0.05)

Table 3: Total score of hepatocyte damage from Post-Hoc

test.

 K1 K2 K3 K4 K5

K1  0.011* 0.493 0.674 0.567

K2 0.011*  0.003* 0.027* 0.036*

K3 0.493 0.003*  0.276 0.218

K4 0.674 0.027* 0.276  0.878

K5 0.567 0.036* 0.218 0.878 

4 DISCUSSION

This research shows that D-Galactose induces cell

death in the liver. The highest degree of liver damage

was found in the D-Galactose-induced group with no

treatment (neither AQMC leaf extract nor vitamin C).

The administration of AQMC leaf extract provides

improved features of hepatic architecture in that the

dose of 35mg/kgBW mirrored the normal features

while the dose of 70 mg/KgBW approximated the

features in the group given vitamin C. The total score

of liver damage was significantly different in all study

groups. A significant difference was particularly

evident between the group with AQMC leaf extract at

a dose of 35 mg/kg BW and the positive control

group. There was an insignificant difference between

the hepatic histological features of the mice receiving

AQMC leaf extract and those of the healthy group.

Therefore, it indicates that the administration of

AQMC leaf extract (especially at a dose of

35mg/kgBW) can improve the histological features of

aging liver (D-galactose-induced). Such histological

changes have the same features with those of the liver

of healthy mice (not induced by D-galactose).

When induced to the body, D-Galactose increases

the levels of galactose, and galactose will reduce to

galactitol, a compound that cannot be further

metabolized, thereby accumulating intracellularly

and increasing the cell osmotic pressure. This process

will eventually lead to swollen cells.

Histologically, this feature is recognized as

ballooning degeneration/cell swelling with clear

cytoplasm (Ye et al., 2014). D-Galactose will also

reduce free amine groups in amino acids or proteins,

thus leading to AGEs formation through glycation as

well as ROS formation (Parameshwaran et al., 2010).

The administration of D-galactose for 6 weeks

correspondingly led to an elevated level of free

radicals (plasma MDA levels) by approximately 3-

fold of that of the healthy group (Sulistyoningrum et

al., 2019).

Muntingia calabura has a hepatoprotective effect.

A study shows that the administration of methanol

extract of Muntingia calabura leaves (MMC leaf

extract) followed by soft drink administration can

significantly prevent increases in SGOT and SGPT

levels (Siddiq et al., 2019). MMC leaf extract

administered for 7 days followed by the

administration of hepatotoxic substances (CCL4 or

paracetamol) can prevent further liver damage

(Zakaria et al., 2019) (Mahmood et al., 2014). MMC

leaf extract as premedication for CCl4 administration

is able to prevent elevated ALT levels, increased pro-

inflammatory cytokines (NO, TNF-α, IL-β, IL-6),

and higher ratio of the liver weight to the body

weight. MMC leaf extract administered to rats at

therapeutic doses of 250 mg/kgBW and 500

mg/KgBW results in a liver weight ratio closer to that

of the healthy group. ALT levels in the 500

mg/KgBW treatment group are similar to those of the

positive control group (receiving N-acetyl-cysteine

therapy and induced by CCl4). MMC leaf extract

(doses of 50.250 and 50 mg/kgBW) as premedication

before induction by CCl4 is also able to significantly

increase the antioxidant levels (SOD and CAT) in the

body. Histopathological features of liver damage

become minimal in the 500 mg/kgBW treatment

group (Zakaria et al., 2019). MMC leaf extract as

premedication is also able to prevent liver damage

induced by paracetamol. In line with the report by

Zakaria et al., (2019), MMC leaf extract as

premedication is able to prevent an increase in the

relative weight of the liver but unable to prevent

increased levels of liver enzymes (ALT, AST, and

ALP). Minimal necrosis and inflammation are found

in the group receiving MMC leaf extract as

premedication (Mahmood et al., 2014).

The doses of 35 mg/KgBW and 70 mg/KgBW for

the mice in this study are equivalent to the doses of

250 mg/KgBW and 500 mg/KgBW in experimental

rats. This indicates that the findings of this study are

in accordance with previous studies in which the

potency of MMC leaf extract is found at both doses

(Mahmood et al., 2014; Siddiq et al., 2019; Zakaria et

al., 2019). In contrast to the potential dose of MMC

leaf extract at 500 mg/kgBW, that of AQMC leaf

extract in this study is found at 250 mg/kgBW. This

The Effects of Aqueous Extract of Jamaican Cherry (Muntingia calabura) on D-galactose-induced Liver Damage in BALB/c Mice (Mus

musculus)

205

difference is likely caused by the different treatment

given to the experimental animals. The main

objective of this study was to examine the therapeutic

effects of AQMC leaf extract on damaged liver while

the study using MMC leaf extract aimed to

investigate the protective effects on the induction of

liver damage. Another reason for the difference is the

compound concentrations in MC due to the extraction

process. Kolar, Kamble and Dixit, (2011) report that

AQMC leaf extract has a total flavonoid content

exceeding that of MMC leaf extract but with a lower

phenolic content. The antioxidant activity of MMC

leaf extract is apparently higher than that of AQMC

leaf extract.

AQMC leaf extract has more benefits than other

types of extract. Compared to petroleum ether and

ethyl acetate extracts, the total phenolic content of

AQMC leaf extract is twice higher with a better

hepatoprotective effect (indicating only minimal

inflammation). In addition, premedication using

AQMC leaf extract is able to suppress increasing

hepatic enzyme levels (ALT, AST, and ALP) in rats

induced by paracetamol. By comparison with the

doses of 50 and 250 mg/KgBW, the dose of 500

mg/KgBW in rats shows the best potency (in terms of

the liver weight ratio, hepatic histological features,

antioxidant levels, and liver enzyme levels) (Zakaria

et al., 2018).

The protective effect of MC is associated with its

antioxidant activities in the flavonoid compounds,

such as catechin, gallocatechin, epigallocatechin

narigenin, and quercetin in MC (Pereira et al., 2018).

The hydroxyl complex in the phenol compounds in

MC can inhibit proton donation in ROS formation

(Balakrishnan et al., 2011). The polyphenol

compounds in MC inhibit glycosidation reactions and

have an anti-glycation activity by inhibiting RAGEs

signaling (Sadowska-Bartosz & Bartosz, 2015). The

saponins in MC also slow down aging through

activation of the AKT FOXO3a pathway and the

Nuclear factor-erythroid 2-related factor-2 pathway.

This process will improve the expression and

functions of antioxidant enzymes, such as superoxide

dismutase-2 (SOD-2), catalase, glutathione

reductase, glutamate-cysteine ligase, and heme

oxygenase-1 (Khan Y et al., 2015). In addition to the

antioxidant potential, research by Rofiee et al., (2015)

shows that the protective effect of MC is manifested

through the bile acid biosynthesis and arachidonic

acid metabolism.

This study has a limitation in that the effects of

AQMC leaf extract on impaired liver function (levels

of ALT, AST and ALP enzymes) were not

investigated. However, the findings of this study have

been able to illustrate that MMC leaf extract has a

therapeutic effect on liver damage, particularly on an

aging liver. Further research with reference to the

therapeutic effects of AQMC leaf extract is required

by taking into account the various stimuli of both

acute and chronic tissue damage.

5 CONCLUSION

The administration of AQMC leaf extract can reduce

liver and hepatocyte damage of BALB/c Mus

musculus induced by D-Galactose. The potential dose

of AQMC leaf extract is 35 mg.

REFERENCES

Aziz Ibrahim, I. A. (2012). Leaves Extract of Muntingia

Calabura Protects Against Gastric Ulcer Induced by

Ethanol in Sprague-Dawley Rats. Clinical and

Experimental Pharmacology, 01(S5).

https://doi.org/10.4172/2161-1459.s5-004

Balakrishnan, K. P., Narayanaswamy, N., & Duraisamy, A.

(2011). Tyrosinase inhibition and anti-oxidant

properties of Muntingia calabura extracts: In vitro

studies. International Journal of Pharma and Bio

Sciences, 2(1), 294–303.

Biondo-Simões, M. D. L. P., Matias, J. E. F., Montibeller,

G. R., Siqueira, L. C. D., Nunes, E. D. S., & Grassi, C.

A. (2006). Effect of aging on liver regeneration in rats.

Acta Cirurgica Brasileira, 21(4), 197–202.

https://doi.org/10.1590/S0102-86502006000400002

Bo-Htay, C., Palee, S., Apaijai, N., Chattipakorn, S. C., &

Chattipakorn, N. (2018). Effects of d-galactose-induced

ageing on the heart and its potential interventions.

Journal of Cellular and Molecular Medicine, 22(3),

1392–1410. https://doi.org/10.1111/jcmm.13472

Hadzi-Petrushev, N., Stojkovski, V., Mitrov, D., &

Mladenov, M. (2015). D-galactose induced changes in

enzymatic antioxidant status in rats of different ages.

Physiological Research, 64(1), 61–70.

https://doi.org/10.33549/physiolres.932786

Hashish, H. A. (2016). Effect of age on the sildenafil impact

on the histological and ultra-structure of the liver in

male albino rat. Journal of Histology and

Histopathology, 3(1), 5. https://doi.org/10.7243/2055-

091x-3-5

Khan Y, M. A., Mundasada, S. C., & Ramadas, D. (2015).

Antioxidant Activity : Root, Leaves and Fruits Aqueous

Extracts of MuntingiaCalabura. Journal of Innovations

in Pharmaceuticals and Biological Sciences, 2(4), 363–

368.

Kolar, F. R., Kamble, V. S., & Dixit, G. B. (2011).

Phytochemical constituents and antioxidant potential of

some underused fruits. African Journal of Pharmacy

JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )

206

and Pharmacology, 5(18), 2067–2072.

https://doi.org/10.5897/AJPP11.475

Lalihatu, M. R., & Sudharmono, U. (2019). Effectiveness

of Boiled Cherry Leaf (muntingia calabura L) and Figs

Leaves (Ficus Carica) Toward SGOT SGPT Serum of

Male Wistar Strain Rats with Acute Hepatitis Models.

Abstract Proceedings International Scholars

Conference, 7(1), 716–726.

https://doi.org/10.35974/isc.v7i1.2076

Lin, J. T., Chang, Y. Y., Chen, Y. C., Shen, B. Y., & Yang,

D. J. (2017). Molecular mechanisms of the effects of

the ethanolic extract of Muntingia calabura Linn. fruit

on lipopolysaccharide-induced pro-inflammatory

mediators in macrophages. Food and Function, 8(3),

1245–1253. https://doi.org/10.1039/c6fo01735e

Mahmood, N. D., Mamat, S. S., Kamisan, F. H., Yahya, F.,

Kamarolzaman, M. F. F., Nasir, N., Mohtarrudin, N.,

Tohid, S. F. M., & Zakaria, Z. A. (2014). Amelioration

of paracetamol-induced hepatotoxicity in rat by the

administration of methanol extract of Muntingia

calabura L. Leaves. BioMed Research International,

2014. https://doi.org/10.1155/2014/695678

Murti, F. K., Amarwati, S., & Wijayahadi, N. (2016).

Terhadap Gambaran Mikroskopis Hepar Tikus Wistar.

Kedokteran Diponegoro, 5(4), 871–883.

Parameshwaran, K., Irwin, M. H., Steliou, K., & Pinkert, C.

A. (2010). D-galactose effectiveness in modeling aging

and therapeutic antioxidant treatment in mice.

Rejuvenation Research, 13(6), 729–735.

https://doi.org/10.1089/rej.2010.1020

Pereira, G. A., Arruda, H. S., de Morais, D. R., Eberlin, M.

N., & Pastore, G. M. (2018). Carbohydrates, volatile

and phenolic compounds composition, and antioxidant

activity of calabura (Muntingia calabura L.) fruit. Food

Research International, 108(March), 264–273.

https://doi.org/10.1016/j.foodres.2018.03.046

Rofiee, M. S., Yusof, M. I. M., Abdul Hisam, E. E., Bannur,

Z., Zakaria, Z. A., Somchit, M. N., Teh, L. K., & Salleh,

M. Z. (2015). Isolating the metabolic pathways

involved in the hepatoprotective effect of Muntingia

calabura against CCl4-induced liver injury using

LC/MS Q-TOF. Journal of Ethnopharmacology, 166,

109–118. https://doi.org/10.1016/j.jep.2015.03.016

Sadowska-Bartosz, I., & Bartosz, G. (2015). Prevention of

protein glycation by natural compounds. Molecules,

20(2), 3309–3334.

https://doi.org/10.3390/molecules20023309

Safrida, S., & Sabri, M. (2019). Effect of Muntingia

calabura l. Stem bark extracts on uric acid concentration

and renal histopathology in diabetic rats. Medicina

(Lithuania), 55(10).

https://doi.org/10.3390/medicina55100695

Sarimanah, J., Ketut Adnyana, I., Sukandar, E. Y., &

Kurniati, N. F. (2017). The antirheumatic activity of

Muntingia calabura L. Leaves ethanol extract and its

fraction. Asian Journal of Pharmaceutical and Clinical

Research,

10(1), 84–86.

https://doi.org/10.22159/ajpcr.2017.v10i1.14102

Schmucker, D. L., & Sanchez, H. (2011). Liver

regeneration and aging: A current perspective. Current

Gerontology and Geriatrics Research, 2011.

https://doi.org/10.1155/2011/526379

Siddiq, M. N. A. A., Marliyati, S. A., Riyadi, H., &

Winarsih, W. (2019). Effects of Kersen leaves extract

(Muntingia calabura L.) on SGOT and SGPT levels of

soft drink induced mice. Jurnal Gizi Dan Pangan,

14(2), 69–76.

https://doi.org/10.25182/jgp.2019.14.2.69-76

Sookoian, S., Castaño, G. O., Scian, R., San Martino, J., &

Pirola, C. J. (2016). Heat Shock Protein 27 is down-

regulated in Ballooned Hepatocytes of Patients with

Nonalcoholic Steatohepatitis (NASH). Scientific

Reports, 6. https://doi.org/10.1038/srep22528

Sulistyoningrum, E., Rosmelia, R., Hamid, M. K., &

Nuraini, W. S. T. (2019). Anti-aging effects of

Muntingia calabura leaves extract in D-galactose-

induced skin aging mouse model. Journal of Applied

Pharmaceutical Science, 9(9), 23–29.

https://doi.org/10.7324/JAPS.2019.90904

Ye, Y., Jia, R. R., Tang, L., & Chen, F. (2014). In vivo

antioxidant and anti-skin-aging activities of ethyl

acetate extraction from idesia polycarpa defatted fruit

residue in aging mice induced by D-galactose.

Evidence-Based Complementary and Alternative

Medicine, 2014. https://doi.org/10.1155/2014/185716

Zakaria, Z. A., Mahmood, N. D., Mamat, S. S., Nasir, N.,

& Omar, M. H. (2018). Endogenous antioxidant and

LOX-mediated systems contribute to the

hepatoprotective activity of aqueous partition of

methanol extract of Muntingia calabura L. leaves

against paracetamol intoxication. Frontiers in

Pharmacology, 8(FEB), 1–14.

https://doi.org/10.3389/fphar.2017.00982

Zakaria, Z. A., Mahmood, N. D., Omar, M. H., Taher, M.,

& Basir, R. (2019). Methanol extract of Muntingia

calabura leaves attenuates CCl4-induced liver injury:

possible synergistic action of flavonoids and volatile

bioactive compounds on endogenous defence system.

Pharmaceutical Biology, 57(1), 335–344.

https://doi.org/10.1080/13880209.2019.1606836

Zhong, H. H., Hu, S. J., Yu, B., Jiang, S. S., Zhang, J., Luo,

D., Yang, M. W., Su, W. Y., Shao, Y. L., Deng, H. L.,

Hong, F. F., & Yang, S. L. (2017). Apoptosis in the

aging liver. Oncotarget, 8(60), 102640–102652.

https://doi.org/10.18632/oncotarget.21123

Zulfi, Z., Ilyas, S., & Hutahaean, S. (2013). Pengaruh

Pemberian Vitamin C Dan E Terhadap Gambaran

Histologis Ginjal Mencit (Mus Musculus L.) Yang

Dipajankan Monosodium Glutamat (Msg). Saintia

Biologi, 1(3), 1–6.

The Effects of Aqueous Extract of Jamaican Cherry (Muntingia calabura) on D-galactose-induced Liver Damage in BALB/c Mice (Mus

musculus)

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