176 Abdul Hamid et al.

/ J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185

Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)

ISSN 1673-1581 (Print); ISSN 1862-1783 (Online)
www.zju.edu.cn/jzus; www.springerlink.com
E-mail: [email protected]

Nephroprotective effects of Zingiber zerumbet Smith

ethyl acetate extract against paracetamol-induced
nephrotoxicity and oxidative stress in rats

Zariyantey ABDUL HAMID1, Siti Balkis BUDIN†‡1, Ng WEN JIE1,

Asmah HAMID1, Khairana HUSAIN2, Jamaludin MOHAMED1
(1Department of Biomedical Science, Faculty of Health Sciences, Universiti Kebangsaan Malaysia,
Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia)
(2Drug and Herbal Research Center, Faculty of Pharmacy, Universiti Kebangsaan Malaysia,
Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia)
†
E-mail: [email protected]
Received Apr. 21, 2011; Revision accepted Aug. 30, 2011; Crosschecked Feb. 17, 2012

Abstract: Paracetamol (PCM) overdose can cause nephrotoxicity with oxidative stress as one of the possible
mechanisms mediating the event. In this study, the effects of ethyl acetate extract of Zingiber zerumbet rhizome
[200 mg per kg of body weight (mg/kg) and 400 mg/kg] on PCM-induced nephrotoxicity were examined. Rats were
divided into five groups containing 10 rats each. The control group received distilled water while other groups were
treated with extract alone (400 mg/kg), PCM alone (750 mg/kg), 750 mg/kg PCM+200 mg/kg extract (PCM+
200-extract), and 750 mg/kg PCM+400 mg/kg extract (PCM+400-extract), respectively, for seven consecutive days.
The Z. zerumbet extract was given intraperitoneally concurrent with oral administration of PCM. Treatment with Z.
zerumbet extract at doses of 200 and 400 mg/kg prevented the PCM-induced nephrotoxicity and oxidative impair-
ments of the kidney, as evidenced by a significantly reduced (P<0.05) level of plasma creatinine, plasma and renal
malondialdehyde (MDA), plasma protein carbonyl, and renal advanced oxidation protein product (AOPP). Furthermore,
both doses were also able to induce a significant increment (P<0.05) of plasma and renal levels of glutathione (GSH)
and plasma superoxide dismutase (SOD) activity. The nephroprotective effects of Z. zerumbet extract were confirmed
by a reduced intensity of renal cellular damage, as evidenced by histological findings. Moreover, Z. zerumbet extract
administered at 400 mg/kg was found to show greater protective effects than that at 200 mg/kg. In conclusion, ethyl
acetate extract of Z. zerumbet rhizome has a protective role against PCM-induced nephrotoxicity and the process is
probably mediated through its antioxidant properties.

Key words: Zingiber zerumbet, Antioxidant, Oxidative stress, Nephrotoxicity, Paracetamol

doi:10.1631/jzus.B1100133 Document code: A CLC number: R965; R285.5

1 Introduction worldwide (Gunnell et al., 2000), and its overdose is

commonly associated with hepatic (Nelson, 1995)
Paracetamol (PCM) or acetaminophen was first and renal damages (Placke et al., 1987). PCM toxicity
discovered in 1889 and is a widely used nonprescrip- is mediated by the activity of its reactive metabolite
tive analgesic and antipyretic agent (Brown, 1968). known as N-acetyl-p-benzoquinoneimine (NAPQI),
PCM toxicity is one of the major causes of poisoning which is detoxified by intracellular glutathione (GSH)
(Borne, 1995). Therefore, an overdose of PCM will
‡
Corresponding author
saturate the conjugation pathways of GSH and cause
© Zhejiang University and Springer-Verlag Berlin Heidelberg 2012 depletion of cellular GSH. This subsequently led to a
 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185 177

reduced capacity of GSH to detoxify NAPQI. In- termined by measuring the levels of plasma creatinine
creased level of NAPQI mediates oxidative damage, (indicator of renal function), endogenous antioxidants
and thus enhances cellular injuries and organ dys- [GSH and superoxide dismutase (SOD)], and oxida-
function, including renal damage (Hart et al., 1994). tive stress markers [malondialdehyde (MDA), ad-
Although the occurrence of nephrotoxicity is vanced oxidation protein product (AOPP), and pro-
less common than that of hepatotoxicity as reported tein carbonylation], and by histological change
by Blakely and McDonald (1995), a number of stu- analysis.
dies have documented that PCM-induced renal tubu-
lar damage and acute renal failure can occur even in
the absence of liver damage (Carpenter and Mudge, 2 Materials and methods
1981), and can be the primary manifestation of PCM
2.1 Plant materials
toxicity (Boutis and Shannon, 2001). As PCM over-
dose is shown to mediate severe renal damage, which Fresh rhizomes (7 kg) of Z. zerumbet were col-
can be life-threatening, antidotes or treatments that lected from Temerloh, Pahang, Malaysia and authen-
could offer maximum protection against PCM- ticated by a plant taxonomist at Department of Botany,
induced renal injury are therefore required. To date, Faculty of Science and Technology, Universiti Ke-
N-acetyl-cysteine (NAC) has been used clinically for bangsaan Malaysia (UKM), and were deposited as a
the treatment of PCM toxicity (Dilger and Baker, voucher specimen at the herbarium of UKM, Bangi,
2007). Selangor, Malaysia. The specimen was cleaned and
Current evidence suggests that PCM-induced chopped into small pieces and then air-dried at room
hepatorenal injury is mediated by oxidative damage temperature for three days.
(Das et al., 2010; Kheradpezhouh et al., 2010).
2.2 Plant extraction
Therefore, the alternative treatments for PCM toxicity
could be achieved using a natural compound with The air-dried rhizomes of Z. zerumbet were se-
antioxidant activity. Zingiber zerumbet (L.) Smith, or quentially soaked at room temperature in n-hexane,
locally known as lempoyang or wild ginger, belong- ethyl acetate, and then methanol for 72 h. The resul-
ing to the Zingiberaceae family (Saadiah and Halijah, tant extracts were filtered and evaporated to dryness
1995) has been shown to possess a number of bio- in vacuo to yield crude extracts of hexane, ethyl ace-
logical activities, including anti-cancer (Huang et al., tate, and methanol. All the crude extracts were stored
2005; Rashid et al., 2005; Sharifah Sakinah et al., at 4 °C until tested for bioassay. Prior to use, the Z.
2007; Abdul et al., 2008), anti-inflammatory (Jaga- zerumbet ethyl acetate extract was dissolved in di-
nath and Ng, 2000; Somchit and Shukriyah, 2003), methyl sulfoxide (DMSO) and diluted in phosphate
antimicrobial (Abdul et al., 2008; Kaderi et al., 2010), buffer saline (PBS; pH 7.4).
and antioxidant properties (Ruslay et al., 2007). Z.
2.3 Experimental protocol
zerumbet has been shown to contain flavonoid com-
pounds that exhibit the antioxidant properties (Pietta, All procedures involving the use of laboratory
2000), and the ethyl acetate extract of the plants has animals were reviewed and approved by the Animal
been shown to exhibit strong antioxidant activities Ethics Committee of UKM. Fifty male Sprague-
(Ruslay et al., 2007). In a recent study, zerumbone, Dawley rats (230–250 g) were obtained from the
which is the active compound of the Z. zerumbet UKM Animal Resource Unit. The animals were
rhizome, has been shown to protect against cispla- housed in a controlled environment with room tem-
tin-induced renal dysfunction by preventing lipid perature and a 12-h light-dark cycle. Animals were
peroxidation and preserving antioxidant (Ibrahim et fed mouse pellet and fresh water ad libitum for a week
al., 2010). prior to experiments. Rats were randomly divided into
In this study, we determined the potential pro- five groups containing 10 animals each and all
tective effects and antioxidant activities of ethyl ace- treatments were given daily for seven days. PCM was
tate extract of the Z. zerumbet rhizome against administered orally, while Z. zerumbet extracts at 200
PCM-induced nephrotoxicity. The effects were de- and 400 mg per kg of body weight (mg/kg) were
178 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185

delivered intraperitoneally concurrent with PCM GSH was quantified using 5,5′-dithiobis-2-
administration. The selected doses of Z. zerumbet nitrobenzoic acid (DTNB) according to the protocols
extract were based on Hemabarathy et al. (2009) who as described by Ellman (1959). The reaction between
showed the hepatoprotective effects of aqueous ex- GSH and DTNB formed a yellow-colored complex
tracts of Alpina Galanga (Zingiberaceae family) on that was measured spectrophotometrically at 420 nm.
the PCM-induced hepatotoxicity. Rats in Group I MDA was determined based on the production
served as the control group and were administered of thiobarbituric acid reactive substances (TBARS),
distilled water only. Groups II received 400 mg/kg Z. the absorbance of which was measured spectropho-
zerumbet extract alone, while Group III received tometrically at 532 nm (Esterbauer and Cheeseman,
750 mg/kg PCM alone. In Group IV, rats were treated 1990). The concentration of TBARS was determined
with 750 mg/kg PCM and 200 mg/kg Z. zerumbet from the extrapolated standard curve of serially di-
extract. Meanwhile, rats in Group V were treated with luted 1,1,3,3-tetraethoxypropane (TEP). The final
750 mg/kg PCM and 400 mg/kg Z. zerumbet extract. concentration of MDA was then expressed in nmol/g
On Day 8, all animals were weighed and anaesthe- protein.
tized with diethyl ether. Protein carbonyls were estimated according to
the procedure as described by Levine et al. (1990)
2.4 Sample preparation
based on the reaction of carbonyl compounds with
Blood was collected via cardiac puncture using a 2,4-dinitrophenyl hydrazine (2,4-DNPH) for 1 h and
10 ml syringe and 25G needle after 24 h of last precipitations with 20% trichloroacetic acid (TCA,
treatment. Blood was collected in EDTA tubes and 0.2 kg/L). The released carbonyl compounds were
plasma was obtained after centrifugation at measured by spectrophotometer at 380 nm. The
3 000 r/min for 10 min at 4 °C. Plasma sample was amounts of carbonyls are calculated based on the total
stored at −20 °C until further analysis. The kidney amount of protein contents in each sample.
was obtained after the rat was sacrificed, washed in AOPP of renal homogenate was measured ac-
ice-cold 11.5 g/L KCl to remove blood, and quickly cording to Witko-Sarsat et al. (1996). Renal homo-
blotted and weighed on digital scale (B303 Mettler- genate was centrifuged at 2 500 r/min for 10 min at
Toledo, Switzerland). For histological studies, the 40 °C. The supernatant was then added to a reaction
kidney was preserved in 10% formalin while for bi- mixture containing 50% acetic acid and 1.16 mol/L
ochemical analysis; the kidney was minced into small potassium iodide (KI) in PBS solution. The absor-
pieces and homogenate with 11.5 g/L KCl in 4 ml/g bance was recorded at 340 nm and the concentration
ratio using homogenizers (Ultra Turrax T25). of AOPP was determined from the extrapolated
standard curve of serially diluted AOPP standard
2.5 Biochemical analysis
solution using 500 μmol/L chloramines stock.
Creatinine level was determined according to
2.6 Histological analysis
Jaffe’s reaction, based on creatinine reaction with
picric acid in alkaline solution to form a red solution Examination of renal histology was performed
measured spectrophotometrically at 520 nm (Slot, according to routine histology techniques. Briefly,
1965). The level of creatinine was extrapolated from after the animal was sacrificed, the kidney was har-
the standard graph and the values were then stated as vested, rinsed in normal saline, and sectioned into
milligram per deciliter (mg/dl). small pieces. The sectioned tissue was then fixed in
SOD activity was assayed according to the me- 10% formalin, dehydrated in stepwise with increasing
thod as described by Beyer and Fridovich (1987) concentration of ethanol solution (50% to 100%), and
based on the reaction of SOD with nitrotetrazolium embedded in paraffin. Using a microtome, tissue
blue chloride (NBT·2HCl), Triton-X 100 and ribof- sections of 4-µm thickness were produced, fixed
lavin and the absorbance was measured with a spec- overnight on the slide, subsequently stained with
trophotometer at 560 nm. One unit (1 U) of SOD was hematoxylin and eosin (H&E), and were then ob-
defined as the amount of SOD required for 50% in- served under a light microscope (Olympus BX41,
hibition of NBT·2HCl reduction. Japan).
 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185 179

2.7 Statistical analysis

Statistical analysis was done using SPSS version
18.0. Data were analyzed using Shapiro-Wilk nor-
mality test and one way analysis of variance
(ANOVA) was used for comparison between groups
followed by post-hoc Tukey test. Data were expressed
as mean±standard deviation (SD) and P<0.05 showed
statistically significance.

3 Results
3.1 Effects of Z. zerumbet extract on body weight,
kidney weight, and food and water intake
In all experimental groups, no mortality was
observed during the period of the study. Reduced
water intake and food consumption, as well as re- Fig. 1 Effects of ethyl acetate extract of Z. zerumbet on
duced body weight, were observed in PCM group as body weight (a) and kidney weight (b) of rats after seven
compared to control (Fig. 1a). In contrast, groups days of treatments
receiving PCM concurrent with administration of Each column represents mean±SD (n=10). a P<0.05 versus
control group; b P<0.05 versus PCM group. 400-extract,
either 200 or 400 mg/kg of Z. zerumbet extract
PCM, PCM+200-extract, and PCM+400-extract groups
weighed significantly higher (P<0.05) than PCM were treated with 400 mg/kg extract alone, 750 mg/kg PCM
group. Meanwhile, no significant alteration of body alone, 750 mg/kg PCM+200 mg/kg extract, and 750 mg/kg
weight was recorded from rats treated with 400 mg/kg PCM+400 mg/kg extract, respectively
of extract only. Furthermore, the kidney weight was
significantly lower (P<0.05) in all treated groups as
compared to the control, and the reduction was mar-
kedly noticeable in the PCM group (Fig. 1b).
3.2 Effect of Z. zerumbet extract on creatinine level
A significantly increased (P<0.05) level of
plasma creatinine was observed in plasma samples of
the PCM group (Fig. 2). However, supplementation
with Z. zerumbet extract at 200 and 400 mg/kg sig-
nificantly prevented (P<0.05) further elevations of Fig. 2 Effects of ethyl acetate extract of Z. zerumbet on
creatinine, with obvious effects observed in the plasma creatinine in rats after seven days of treatments
Each column represents mean±SD (n=10). a P<0.05 versus
PCM+400 mg/kg extract group. There were no
control group; b P<0.05 versus PCM group; c P<0.05 versus
nephrotoxic effects observed in rats administered PCM+200-extract group
400 mg/kg of the Z. zerumbet extract alone, as evi-
denced by no significant difference in plasma creati- 200 or 400 mg/kg of Z. zerumbet extract significantly
nine levels as compared to the control group. prevented (P<0.05) PCM-induced GSH reductions,
with greater effects demonstrated in the PCM+
3.3 Effect of Z. zerumbet extract on antioxidant
400 mg/kg extract group, which was able to maintain
status
the level of GSH despite the presence of PCM. Fur-
Administration of PCM for seven consecutive thermore, supplementation of 400 mg/kg of Z. ze-
days caused a significant reduction (P<0.05) of GSH rumbet extract alone showed no significant difference
level in both plasma and renal homogenate (Figs. 3a in plasma and renal levels of GSH as compared to the
and 3b). In contrast, administrations with either control group.
180 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185

Fig. 3 Effects of ethyl acetate extract of Z. zerumbet on the levels of plasma GSH (a), renal GSH (b), plasma SOD
activity (c), and renal SOD activity (d) in rats after seven days of treatments
Each column represents mean±SD (n=10). a P<0.05 versus control group; b P<0.05 versus PCM group; c P<0.05 versus
PCM+200-extract group

In addition to the GSH level, the activity of SOD and Z. zerumbet extract at 200 and 400 mg/kg,
enzymes in plasma and renal tissue was also deter- however, reduced the oxidative stress, as evidenced
mined. The group treated with PCM alone showed by significantly reduced (P<0.05) level of renal ho-
significantly lower (P<0.05) plasma SOD activity as mogenate and plasma MDA, plasma protein carbonyl,
compared to the control (Fig. 3c). Administration of and renal AOPP than those in the PCM group. Inte-
extracts at both doses significantly increased (P<0.05) restingly, administration of both doses brought the
plasma SOD activity than the PCM group. In addition, MDA level of renal homogenate nearly to the value of
supplementation with 400 mg/kg extract together the control group. In addition, no significant induc-
with PCM further increased plasma SOD activity than tion of oxidative stress was observed in rats admi-
the PCM+200 mg/kg extract groups, and its plasma nistered with 400 mg/kg of the extract alone, as evi-
SOD activity was almost similar as that in the control denced by no significant difference in the value of
group. In contrast to the activity of plasma SOD, the renal homogenate and plasma MDA, plasma protein
renal homogenate SOD activity did not show any carbonyl, and renal AOPP than in control.
significant difference among all experimental groups
3.5 Effect of Z. zerumbet extract on histological
(Fig. 3d).
features of the kidney
3.4 Effect of Z. zerumbet extract on oxidative stress
The histological features of the kidneys from
Administration of PCM for seven consecutive various treatments groups are as presented in Fig. 5.
days induced oxidative stress with a significant in- Renal sections from the control and 400 mg/kg Z.
crease (P<0.05) in the levels of renal homogenate and zerumbet group showed no histological changes, as
plasma MDA, plasma protein carbonyl, and renal evidenced by the normal appearance of glomeruli and
AOPP (Fig. 4). Concurrent administration of PCM tubules (Figs. 5a and 5b). In contrast, kidney sections
 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185 181

Fig. 4 Effects of ethyl acetate extract of Z. zerumbet on the levels of plasma MDA (a), renal MDA (b), plasma protein
carbonyl (c), and renal AOPP (d) in rats after seven days of treatments
Each column represents mean±SD (n=10). a P<0.05 versus control group; b P<0.05 versus PCM group, c P<0.05 versus
PCM+200-extract group

Fig. 5 Kidney sections of each group (hematoxylin and eosin stained)

(a) Control group showing normal appearance of glomeruli (G) and tubules (T); (b) 400 mg/kg extract group exhibiting
normal features of glomeruli (G) and tubules (T); (c) PCM group showing severe glomerular (G) and tubular damages (T);
(d) PCM+200 mg/kg extract group showing moderate glomerular (G) and tubular destruction (T); (e) PCM+400 mg/kg
extract group showing almost normal appearance of glomeruli (G) and tubules (T)
182 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185

from the PCM group exhibited altered architecture lowitz, 2000). In our study, administration of
with extensive destruction of glomeruli and tubular 750 mg/kg PCM caused significant elevations of
structures, as demonstrated by marked necrotic areas plasma and renal MDA. In contrast, treatment with
(Fig. 5c). On the other hand, administration of Z. ze- ethyl acetate extract of Z. zerumbet abrogated this
rumbet extract at 200 (Fig. 5d) or 400 mg/kg (Fig. 5e) elevation with greater protection being shown with
concurrent with PCM administration, showed pro- the 400 mg/kg extract. The ability of Z. zerumbet to
tective effects on the kidney, as evidenced by the less protect lipid peroxidation is in agreement with Ruslay
severe tubular and glomerular damages, with better et al. (2007), who demonstrated that ethyl acetate
protection being observed in the PCM+400 mg/kg fractions of Z. zerumbet possess remarkable lipid
extract group. These histological observations were in peroxidation inhibition and radical scavenging activ-
line with biochemical findings and all these results ities. Pietta (2000) also showed that the ethyl acetate
support each other. fraction of Z. zerumbet contains flavonoid glycosides
known as afzelin and diacetylafzelin, and derivatives
of kaempferol, a 3-flavonol that was known to pos-
4 Discussion sess antioxidant property. In addition, Ibrahim et al.
(2010) reported that pretreatment with zerumbone, a
PCM-induced nephrotoxicity has been pre- natural compound isolated from fresh rhizomes of Z.
viously documented by a number of studies (Mitchell zerumbet protected against cisplatin-induced neph-
et al., 1977; McMurtry et al., 1978; Newton et al., rotoxicity by attenuating the production of MDA and
1983). In this study, we demonstrated that the ad- simultaneously promoting GSH production. There-
ministration of 750 mg/kg PCM for seven consecu- fore, we believe that the effects manifested by the
tive days was able to induce nephrotoxicity in rats, as crude ethyl acetate extract of Z. zerumbet to overcome
demonstrated by the significantly increased level of PCM-induced nephrotoxicity in our study were me-
plasma creatinine. The nephrotoxic effect of PCM diated by the presence of these bioactive compounds.
was further confirmed by the severely destructed Excess reactive oxygen species (ROS) can
renal tissue, as shown in the histological analysis. promote protein oxidation, forming protein carbonyls
This finding is in accordance with that of Abdel- (Chevion et al., 2000; Margetis et al., 2009) and
Zaher et al. (2007) who showed that the administra- AOPP, which have been described as reliable markers
tion of PCM at 750 mg/kg induced nephrotoxicity in to estimate the degree of oxidant-mediated protein
rats. Administration of ethyl acetate extract of Z. damage (Alderman et al., 2002). The significant
zerumbet concurrent with PCM exposure prevented elevation of plasma protein carbonyls and renal ho-
PCM-induced nephrotoxicity. Furthermore, the pro- mogenate AOPP levels in the PCM group indicates
tective effect was found to be more remarkable in the the presence of oxidative stress and upregulation of
PCM+400 mg/kg extract group than in the PCM+ NAPQI production, which stimulates protein oxida-
200 mg/kg extract group. tions. Concurrent delivery of Z. zerumbet extract at
To date, a number of studies have suggested the both doses, however, reduced significantly the pro-
involvement of oxidative stress as the mediator of duction of plasma protein carbonyls and renal ho-
PCM-induced nephrotoxicity (Das et al., 2010; Khe- mogenate AOPP, and this demonstrates the capacity
radpezhouh et al., 2010; Yousef et al., 2010). Hart et of the Z. zerumbet extract to overcome ROS produc-
al. (1994) pointed out that PCM toxicity was shown tion, presumably through its antioxidants property.
to enhance NAPQI production. The accumulated GSH is a well-known non-enzymatic antioxidant
NAPQI induces greater formation of free radicals, that is responsible as the frontier defense mechanism
which are responsible in mediating cellular damages during oxidative stress and the depletion of intracel-
and renal toxicity. lular GSH in a prolonged oxidative stress (Kaplowitz,
Elevated levels of MDA in tissue have been re- 2000). As demonstrated in our findings, a significant
garded as an indicator for cellular damage due to decline in both plasma and renal homogenate GSH
excess lipid peroxidation processes that occur during levels in PCM-treated rats is in line with the previous
malfunction of the antioxidant defense system (Kap- finding that oral administration of 750 mg/kg PCM to
 Abdul Hamid et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2012 13(3):176-185 183

rats for seven days reduced renal cellular contents of ical and histological analyses. However, despite no
GSH (Abdel-Zaher et al., 2007). The administration nephrotoxic evidence, rats administered with
of Z. zerumbet ethyl acetate extract at 200 and 400 mg/kg extract of Z. zerumbet alone showed sig-
400 mg/kg preserved GSH levels and the effects were nificantly reduced kidney weight as compared to
remarkably effective especially in rats received control rats. Therefore, at this stage, we cannot as-
400 mg/kg Z. zerumbet extract. Ibrahim et al. (2010) certain the definitive reason for the above condition;
reported that zerumbone compound in Z. zerumbet however, we suggest that the Z. zerumbet extract,
extract indirectly induces the biosynthesis of GSH when administered at 400 mg/kg, may affect the
and provides a protective intracellular mechanism, kidney without remarkable alteration of renal histol-
presumably as a free radical scavenger for toxic ogy. Thus, the administration of the Z. zerumbet ex-
agents. Hence, we can speculate that the Z. zerumbet tract at 400 mg/kg or higher in rats may possess a risk
extract in our study may induce the synthesis of en- of toxicity, and further study is required to elucidate
dogenous GSH, which protects against PCM-induced this issue.
oxidative damage and greater effects at 400 mg/kg
treated rats are presumably conferred through higher
amounts of zerumbone present in the extracts. 5 Conclusions
Administration of the Z. zerumbet extract at 200
The administration of Z. zerumbet ethyl acetate
and 400 mg/kg was again able to maintain the SOD
extract was found to have protective effects and an-
activity during PCM overdose. Improvement in SOD
tioxidant activities against PCM-induced nephrotox-
activities does provide evidence to an overall im-
icity, as evidenced by the biochemical status and
provement in endogenous antioxidant defense system
histological findings. The most remarkable effects
(Hemabarathy et al., 2009). This relates to the fact
were observed when the Z. zerumbet extract was
that Z. zerumbet has been shown to have high anti-
delivered at 400 mg/kg as compared to a lower dose
oxidant activities through induction of the endogen-
of 200 mg/kg of the extract. This indicates that the
ous antioxidants that reduce free radical activity
amount of antioxidant compounds present in Z. ze-
(Nakamura et al., 2004). In contrast, SOD activities
rumbet contributes significantly to its antioxidant
measured from renal homogenate were not signifi-
property. Together, the absence of renal damage and
cantly different among all groups. This finding is
supportive evidence of its antioxidant properties may
supported by Hemabarathy et al. (2009) who dem-
suggest the potential applications of Z. zerumbet as an
onstrated that no significant alteration of liver SOD
alternative antidote against PCM-induced nephro-
levels was observed during PCM-induced hepato-
toxicity, as well as a novel antioxidant.
toxicity. This finding suggests that GSH is responsi-
ble as the foremost antioxidant defense mechanism
against PCM-induced oxidative damage, and the Acknowledgements
significant alteration of GSH levels will appear before
the depletion of SOD activities. The authors are grateful to the Faculty of Health
As demonstrated in this study, rats exposed to Sciences (FSK), Universiti Kebangsaan Malaysia for
PCM in the absence of the Z. zerumbet extract showed financial assistance. The authors would like to ac-
severe cellular damage. In contrast, concurrent knowledge everyone in the faculty who contributed
treatment with the Z. zerumbet extract managed to directly or indirectly to this research.
protect against severe cellular lesions, with only mild
or moderate degeneration of glomerular and tubular References
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