Biomedicine & Preventive Nutrition 4 (2014) 225230

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Original article

Protective effect of curcumin on chloroform as by-product of water

chlorination induced cardiotoxicity
Afrah F. Salama a , Ehab Tousson b , Kamal A.F. Shalaby c , Hind T. Hussien a,
a
Biochemistry Section, Department of Chemistry, Faculty of Science, Tanta University, Egypt
b
Department of Zoology, Faculty of Science, Tanta University, Egypt
c
Biochemistry Department, Faculty of Science, Ain Shams University, Egypt

a r t i c l e i n f o a b s t r a c t

Article history: Chloroform (CHCl3 ) is one of the volatile organic compounds detected most frequently in both ground and
Received 18 June 2013 surface water. This study aimed to evaluate the efcacy of curcumin (CMN) to attenuate CHCl3 toxicity
Accepted 13 February 2014 and cellular dysfunction in cardiac tissue of female albino rats. Fifty rats were divided into 5 groups,
1st group was control; 2nd group rats were intoxicated with 150 mg CHCl3 /kg BW; 3rd group rats were
Keywords: treated with 50 mg CMN/kg BW; 4th group rats were treated with 50 mg CMN/kg BW for 30 days then
Chloroform intoxicated with 150 mg CHCl3 /kg BW for 60 days and 5th group rats were intoxicated with 150 mg
Curcumin
CHCl3 /kg BW plus 50 mg CMN/kg BW, respectively. Treatment was continued for 90 days. The levels of
Heart
Antioxidant enzymes
lipid peroxidation, myeloperoxidase (MPO) and xanthine oxidase (XO) were increased and the activities
Cardiac toxicity of antioxidant enzymes, protein content and endogenous antioxidants were decreased in cardiac tissues
in rats treated with CHCl3 in comparison with control group. Serum cholesterol, triglycerides and LDL-C
levels were increased while high HDL-C was decreased in rats treated with CHCl3 in comparison with
control group. Treatment with CMN helps in improving the adverse effect of CHCl3 toxicity; also our
histological results conrm this nding. The present study could be concluded that CMN has protective
and ameliorative effects against CHCl3 induced oxidative stress.
2014 Published by Elsevier Masson SAS.

1. Introduction P450-2E1 is the enzyme that catalyses the conversion process.

Chloroform (CHCl3 ) is the most prevalent by-product of water
disinfection with chlorine-based chemicals [1] and is also formed
in large quantities as a by-product of chlorination of cooling water
in power plant and in the process of bleaching paper [2]. There-
fore, a large part of the human population may be chronically
exposed to chloroform from different sources. Also, drinking water
has been considered the main one. This aliphatic organic compound
is representative of a large group of organic compound having a
widespread use in industry as dispersant, solubilizer and diluent.
It is one of the top 20 priority environmental toxicant [3] and is
classied as a group 2B carcinogen [4]. The acute toxicity demon-
strated by CHCl3 is due to its biotransformation to nucleophilic
bi-functional metabolite phosgene that reacts with glutathione
to fom diglutathionyl dithiocarbonate or is directly metabolized
to carbon dioxide and chloride free radicals [5]. Cytochrome
Corresponding author.
E-mail address: hindtarik@yahoo.com (H.T. Hussien).
Phosgene, formed in the biotransformation process of CHCl3 , bind
covalently to tissue macromolecules resulting in impairment of cel-
lular vital functions. CHCl3 like most other liver damaging agents,
such as acetaminophen and alcohol or viral hepatitis release reac-
tive oxygen species (ROS) at the site of injury [6]. The acute toxic
effects of CHCl3 in animals are similar to those observed in humans
and the main target organs are the liver, kidney and the central
nervous system [7].
CHCl3 toxicity is basically mediated by free radical; there-
fore a free radical scavenger such as curcumin has been in
use in CHCl3 insults. Curcumin, an important constituent of
turmeric (Curcuma longa L.), has been widely used for centuries
as an indigenous medicine [8]. Curcumin exhibits a wide range
of pharmacological effects such as antioxidant, antitumor, anti-
inammatory and hepatoprotective effects of curcumin against
1,2-dimethylhydrazine-induced colon cancer and alcohol as well as
carbon tetrachloride induced hepatotoxicity [9]. The assay of lipid
peroxidation and antioxidant enzymes in cardiac tissue of chloro-
form treated animals has evolved as a reliable method for screening
protective effect of curcumin during chloroform induced toxicity in
female rats.

http://dx.doi.org/10.1016/j.bionut.2014.02.004
2210-5239/ 2014 Published by Elsevier Masson SAS.
226 A.F. Salama et al. / Biomedicine & Preventive Nutrition 4 (2014) 225230
2. Materials and methods
2.1. Chemicals
Chloroform (CHCl3 ), curcumin (Curcuma longa L.) and other
ne chemicals were obtained from sigma chemical company St
Louis, U.S.A. All other chemicals and reagents used were of ana-
lytical grade. The doses of chloroform (CHCl3 ) and curcumin were
150 mg/kg BW [10] and 50 mg/kg BW [11].
2.2. Animals
Fifty adult female albino rats (Rattus norvigicus) weighting about
110 10 g were obtained from the breeding unit of Egyptian orga-
nization for biological products and vaccines, Helwan, Egypt. The
rats were kept in the laboratory for one week before the experimen-
tal work and maintained on a standard rodent diet (20% casein, 15%
corn oil, 55% corn starch, 5% salt mixture, and 5% vitaminized starch;
Egyptian Company of Oils and Soap Kafr-Elzayat, Egypt) and water
available ad libitum. Light was on a 12:12 hr light to dark cycle. The
experimental protocol was approved by Local Ethics Committee
and Animals Research.
2.3. Experimental protocol
The rats were randomly and equally divided into 5 groups (10
animals each).
G1: rats were given the vehicle (corn oil) only at a dose volume
of 2.0 ml/kg BW/day by oral gavage for 90 successive days (control
group).
G2: rats were intoxicated with chloroform by gavage at a dose
of 150 mg/kg BW/day dissolved in corn oil for 90 successive days.
G3: rats were treated with curcumin by gavage at dose of
50 mg/kg BW/day for 90 successive days.
G4: rats were treated with curcumin at a dose 50 mg/kg BW/day
by gavage for 30 days then given chloroform at a dose 150 mg/kg
BW/days by gavage for 60 successive days.
G5: rats were co-treated with curcumin at a dose50 mg/kg
BW/day by gavage plus chloroform at a dose 150 mg/kg BW/days
by gavage for 90 successive days.
At the end of experiment, blood samples were individually col-
lected from the inferior vena cava of each rat in heparinized and
non-heparinized glass tubes. Blood serum was separated by cen-
trifugation at 3000 rpm for 15 minutes. The collected serum was
stored at 18 C. The concentration of cholesterol, triglyceride,
high-density lipoprotein-cholesterol (HDL-C) and low density
lipoprotein-cholesterol (LDL-C) were determined with Kits from
ELLTECH.
Heart was immediately removed; washed using chilled saline
solution then weighted. Tissues were minced and homogenized
(10% W/V), separately in ice-cold sodium phosphate buffer (0.01 M,
pH 7.4) containing 1.15% KCl in a potter El Vehjem type homoge-
nizer. The homogenate was centrifuged at 10.000 g for 15 min at
4 C and the resultant supernatant was used for determination of
biochemical parameter.
The activity of MPO was determined by using O diansidine and
hydrogen peroxide according to Buchman and Lindenstrom [12].
XO activity was determined according to the method described
by Waud and Rajagopalan [13]. CAT activity was measured spec-
trophotometrically at 240 nm by calculating the rate of regarding
to H2 O2 the substrate of the enzyme [14]. The glutathione perox-
idase activity (GSH-Px) was measured according to the method of
Gross et al. [15]. According to the method of Mesbash et al. [16], the
extent of lipid peroxidation was measured in the term of thiobar-
bituric acid reactive substance (TBARS) formation was measured.
Tissue total antioxidants capacity were measured by the method of
described by Benzie and Strain [17]. Total thiol content and non pro-
tein thiol (GSH) were performed according to Sedlak and Lindsay
[18]. The protein content of tissues was determined by the method
described by Lowry et al. [19].
Three rats from each group were anesthetized with Thiopental.
The thorax was opened with surgical incision on the sternum and
the perfusion was done from left ventricle and right atrium. A rins-
ing solution was perfused before the xation solution (10% neutral
buffer formalin). To make rinsing solution, 9.0 g NaCl, 25 g polyvinyl
pyrrolidone, 0.25 g heparin, and 5.0 g procain-HCL were dissolved
in one liter of water by thorough stirring. The pH was adjusted
to 7.35 with 1 N NaOH and twice ltered through Millipore lters
of 3.0 m or less pore size. The perfusion of both solutions was
performed by using a scalp vein attached to a 50cc syringe. Hearts
were immediately removed taking care to handle specimens gently
and a portion of left ventricular was xed in 10% neutral buffered
formalin for standard histological processing parafn sections. Sec-
tions were used for haematoxylin and eosin stains as a routine
method after Bancroft and Stevens [20] for general morphological
evaluation.
2.4. Statistical analysis
One-way analysis of variance (ANOVA) was used to assess sig-
nicant difference among treatment groups. For each signicant
effect of treatment, the Dunnett lest was used for comparisons. The
criterion for statistical signicance was net at P 0.05 or P 0.01
[21].
3. Results
The present study was intended to characterize the role of
oxidative stress in the mechanism of CHCl3 toxicity by study-
ing the change in same parameters after sub chronic exposure.
Table 1 shows signicant decrease in body weight gain in group
treated with CHCl3 and group treated with CMN then CHCl3 while
a signicant increase in relative organ weight of heart in both
groups II and IV as compared with the control. Table 2 shows the
changes in lipid prole in different groups under study; serum
cholesterol, LDL-C and TG were signicantly higher in group II
(P < 0.01) and group IV (P < 0.05) as compared with the control.
Mean serum HDL were signicantly lower (P < 0.01) in groups II and
IV as compared with the control. On the contrary, we showed that
curcumin alleviated the effect of CHCl3 in group treated with cur-
cumin plus CHCl3 . Also, curcumin depleted signicantly the level
of cholesterol, TG, LDL-C in group treated with CMN. Table 3 shows
signicant increase of MDA and reduction of protein content in
(P < 0.01) in both groups II, IV as compared with the control. In
contrast, we showed signicant decrease (P < 0.01) in MDA con-
centration of group treated with CMN and signicant as compared
with the control group. On the other hand, we showed insigni-
cant change in protein content of group III as compared with the
control. Total thiol content, GSH content and TAC were signicant
decrease in both groups II, IV in comparison to its correspond-
ing of group I, in the contrast, we showed signicant increase
(P < 0.01) in group treated with CMN as compared with curcumin
(Table 3).
The activities of GSH-Px and CAT in heart of all studied groups
were shown in Table 3, a signicant decrease (P < 0.01) of both
groups II, IV in correspondence with the control. GSH-Px and CAT
enzymes activities account signicant increase in group treated
with CMN as compared with the control. The present results
showed that CHCl3 signicantly elevated (P < 0.01) the activities
of xanthine oxidase and myeloperoxidase in heart of rats treated
with CHCl3 (Table 3). Although the current study reected that the
 A.F. Salama et al. / Biomedicine & Preventive Nutrition 4 (2014) 225230 227

Table 1
Body weight gain (g). Heart relative weights (g/100 g BW) of female albino rats treated with chloroform (CHCl3 ), curcumin (CMN), curcumin then chloroform and combination
between curcumin and chloroform compared to the control group, (n = 10).

Parameters Experimental groups

Control CHCl3 CMN CMN then CHCl3 CMN + CHCl3

BWG 59.1 5.1 5. 1 0.6 **

58.7 3.15 9 0.2 **
50 1
Heart 42 0.01. 0.53 0.02* 0.42 0.02 0.49 0.02* 0.44 0.047

The signicance of difference was analyzed by one-way ANOVA and Dunnett test (compare all vs. control) using computer program. Values are expressed as means SE.
ANOVA was signicant at P < 0.05. Dunnett test was signicant from corresponding control value at * P < 0.05 and ** P < 0.01.

Table 2
Lipoprotein prole of female albino rats treated with chloroform (CHCl3 ), curcumin (CMN), curcumin then chloroform and combination between curcumin and chloroform
compared to the control group, (n = 10).

Lipid prole(mg/dl) Experimental groups

Control CHCl3 CMN CMN then CHCl3 CMN + CHCl3

Cholesterol 71 1.58 110.1 0.9** 45.4 0.57** 80.8 0.8* 72.5 0.71
TG 143 2.9 170 1.4** 137 2.5* 163 1.5* 145 3.8
HDL-C 33.8 0.67 22.1 0.54** 40.4 0.7** 27.1 0.7** 32.2 0.627
LDL-C 27 1.4 36.7 1.4** 19 1.2* 35.5 0.65* 33.3 3.8

The signicance of difference was analyzed by one-way ANOVA and Dunnett test (compare all vs. control) using computer program. Values are expressed as means SE.
ANOVA was signicant at P < 0.05. Dunnett test was signicant from corresponding control value at * P < 0.05 and ** P < 0.01.

activities of XO and MPO in heart of group III were shown signicant
decrease (P < 0.05) in comparison with group I.
Light microscopy of the left ventricle sections in control rat
group (G1) showed normal myobrillar structure with striations,
branched appearance and continuity with adjacent myobrils
(Fig. 1A). Also, rat ventricle sections in curcumin group (G3)
revealed a normal myobrillar structure with striations as in
control group (Fig. 1B). Left ventricle section in intoxicated
with chloroform group (G2) showed many of abnormalities as
hydrophobic changes of myobrillar structure with striations,
myocardial hypertrophy, severe focal necrosis of cardiac myocytes
associated with inammatory cell inltration and focal haemorr-
hage (Fig. 1C and D).
Left ventricle sections in rats that treated with curcumin for 30
days then given chloroform for 60 successive days (G4) revealed
mild tissue injury with mild myocardial hypertrophy and mild focal
necrosis of cardiac myocytes associated with inammatory cell
inltration (Fig. 1E and F). Left ventricle sections in rats that treated
with curcumin and chloroform for 90 successive days (G5) showed
a few tissue injuries with mild focal hemorrhage and myocardial
cell inltration (Fig. 1G and H).
4. Discussion
The present study was carried out to investigate the protective
effects of curcumin (CMN) on chloroform induced oxidative stress
and biochemical alterations in cardiac tissues rats. The results of
the current study indicated signicant decrease in the body weight
gain of group treated with CHCl3 and group treated with CMN
then CHCl3 as compared with control group, while, the presence
of curcumin with CHCl3 alleviated its toxic effects. The result of
the present study showed that elevation in relative organ weight
of heart in animals treated with CHCl3 in comparison with the
control (Table 1). Frey and Olson [22] mentioned that cardiac hyper-
trophy is certainly an adaptive enlargement inside myocardium
(heart muscle) addressing the many stresses which would grad-
ually results in heart failure and loss of life.
The present data indicated that cholesterol; triglycerides and
LDL-C were signicantly increased in CHCl3 treatment group and
group treated with CMN then CHCl3 while HDL-C levels were
decreased and this is in accordance with the results reported
by Ruddick et al. [23] and Revis et al. [24]. We suggested that
CHCl3 exposure lead to disturbance of lipid metabolism and an

Table 3
Changes in the levels of TBARS (nmole/g), T.thiol (mMole/g), GSH (mmole/g), TAC (mole/g) and t.protein (mg/g) and the activities of CAT (mole/min/g), GSH-PX
(mMole/min/g), XO (mole/min/g) and MPO (nmole/min/g) in cardiac tissue of female albino rats treated with chloroform (CHCl3 ), curcumin (CMN), curcumin then chloroform
and combination between curcumin and chloroform compared to the control group, (n = 10).

Parameters Experimental groups

Control CHCl3 CMN CMN then CHCl3 CMN + CHCl3

TBARS 35 0.6 50 0.3 **

30 0.6 **
45 0.1 **
36 0.2
T.thiol 22.3 0.8 13.7 0.9** 26.2 0.9** 15.6 0.8** 21.9 0.6
GSH 18.8 0.5 13.1 1** 22.1 0.8* 15.1 0.6** 17.3 0.8
TAC 1.017 0.02 0.7 0.007** 1.8 0.13** 0.84 0.03* 1.04 0.02
T.protein 32.5 0.5 24.1 0.8** 32.1 0.8 28.6 0.63** 35.2 1
CAT 1.2 0.04 0.8 0.022** 2.2 0.045** 1 0.03** 1.3 0.03
GSH-Px 18.8 0.9 11.4 1.1** 21.9 1* 14.8 0.5** 18.4 0.3
XO 2.7 0.03 4.8 0.3** 2 0.1* 3.8 0.2** 2.8 0.09
MPO 4.5 0.3 6.5 0.4** 3.3 0.1* 5.9 0.3** 4.6 0.04

The signicance of difference was analyzed by one-way ANOVA and Dunnett test (compare all vs. control) using computer program. Values are expressed as means SE.
ANOVA was signicant at P < 0.05. Dunnett test was signicant from corresponding control value at * P < 0.05 and ** P < 0.01. TBARS: thiobarbituric acid reactive substance;
GSH-Px: glutathione peroxidase activity; XO: xanthine oxidase; MPO: myeloperoxidase.
228 A.F. Salama et al. / Biomedicine & Preventive Nutrition 4 (2014) 225230

Fig. 1. AH. Photomicrographs of the cardiac myocytes of rat left ventricle stained by HE. A. Micrograph from control group of the cardiac myocytes of rat left ventricle
displays normal myobrillar structure with striations, branched appearance and continuity with adjacent myobrils. B. Micrograph from curcumin group (G3 ) revealed a
normal myobrillar structure with striations. C and D. Micrographs of the cardiac myocytes in intoxicated with chloroform group (G2 ) showed myocardial hypertrophy and
severe focal necrosis of cardiac myocytes associated with inammatory cell inltration. E and F. Micrographs of the cardiac myocytes in rats that treated with curcumin
and then given chloroform (G4 ) revealed mild tissue injury with mild myocardial hypertrophy and mild focal necrosis of cardiac myocytes associated with inammatory
cell inltration. G and H. Micrographs of the cardiac myocytes in rats that treated with curcumin and chloroform (G5 ) revealed showed a few tissue injury with mild focal
hemorrhage and myocardial cell inltration.
 A.F. Salama et al. / Biomedicine & Preventive Nutrition 4 (2014) 225230
elevation of serum cholesterol. So, reduction in HDL-C and
increased in cholesterol, TG and LDL-C revealed that CHCl3 induced
cardiovascular diseases, myocardial infarction and our histopatho-
logical examination emphasized these results.
XO is nal enzyme in the conversion of hypoxanthine to xan-
thine a subsequently; to uric acid. XO requires the reduction of
molecular oxygen for urine oxidation and generates the superox-
ide anion radical and subsequently other partially reduced oxygen
species [25]. Our results showed that CHCl3 signicantly elevated
the activity of XO in cardiac tissue of rats treated with CHCl3 by
enhancement superoxide anion suggesting its possible role in the
toxicity. Ali et al. [26] reported that the mammalian avin hydrox-
ylases (XO and aldhyde oxidase) catalyze the biotransformation of
CHCl3 and produce ROS while catalyzing the reaction. However,
the nature of ROS implication in CHCl3 toxicity is not yet well
known. Products of lipid peroxidation may lead to change in biolog-
ical membranes, therefore these changes result in serious cellular
injury. An increase was observed in the formation of MDA in the
cardiac tissue of rats which were exposed to CHCl3 . It is suggested
that ROS play a critical role in the accumulation of neutrophil in tis-
sues after ischemia, where activated neutrophil are also a potential
source of ROS. MPO play a basic role in the production of oxidant
by neutrophil [27].
In our study, the tissue damage produced by increasing the
activity of MPO in heart of rats administrated chloroform and in
group treated with the CMN then CHCl3 as compared with the
control.
Ali et al. [26] claimed that halomethanes like CHCl3 increase
the levels of MDA content in liver. Although Abbassi et al. [28]
recorded a slight but statistically signicant increase of MDA in sin-
gle dose of CHCl3 24- and 48 hour groups only at the non-cytotoxic
dose (150 mg/kg) of this halomethanes. Also, we observed deple-
tion of total protein content in heart of group treated with CHCl3
as compared with the control. Organisms may have an endoge-
nous protective antioxidant defend system against the damages
of free radicals SOD, CAT and GSH-Px are enzymatic antioxidants
that catalyze detoxication reactions of toxic oxygen metabolism
[29]. The mentioned damages may be limited by non-enzymatic
antioxidants such as vitamin A, E and C, melatonin, glutathione and
others [30]. GSH seem to be determinant in the CHCl3 detoxica-
tion process. Consequently, the latter could trigger GSH depletion
[31]. The cellular pool of GSH is maintained by the activity sta-
tus off the enzyme glutamyl cycle ligase catalytic subunit (GCLC;
formerly known as GCS), which in turn is regulated by oxidative
stress [32]. According to the present experimental, we showed a
signicant decrease in both GSH and total thiol content in cardiac
tissue of rats treated with CHCl3 and rats treated with CMN then
CHCl3 as compared with the control. The present study contributed
information towards establishing an association between CHCl3
induced oxidative stress and TAC as it declare signicant decrease
in cardiac tissue of group treated with CHCl3 and group treated with
CMN then CHCl3 group. This may be due to increased utilization of
this antioxidant to counter lipid peroxidation. In the current study,
we demonstrated that CHCl3 treatment decreased signicantly CAT
activity of heart in rats treated with CHCl3 and group treated with
CMN then CHCl3 group in comparison with the control. This is in
agreement with Ali et al. [26]. In the contrary, Abbassi et al. [28]
reported that CAT activity was not affected by CHCl3 after sub acute
(exposure 10 day). Our Observation may be explained by the fact
that CAT is sensitive to ROS and that the oxidative stress resulting
from CHCl3 exposure is strong enough damage. Our current study
recorded a signicant decrease in the activity of GSH-Px of heart
in rats treated with CHCl3 and group treated with CMN then CHCl3
group. In accordance with Ali et al. [26], when treated with sin-
gle dose of chloroform (1500 mg/kg BW) and observed depletion
in GSH-Px activity of hepatic cells. This may be explained by its
229
inactivation resulting from the accumulation of ROS as estimated
by the increase of reduced cytochrome c reaching 60% after acute
exposure and previously proposed by Blum and Fridovich [33].
Besides, since GSH is substrate for GSH-Px, GSH depletion is likely
to have led to the accumulation of peroxides which could be trans-
formed to ROS through the fenton reaction [34]. Although certain
compounds have been tested for the detoxication of CHCl3 [28],
there is no previous study carried out with curcumin. In the current
study, treatment with curcumin alone did not change signicant
effects on the body weight gain and relative organ weight of heart
as compared with the control group. This nding is in accordance
with Asai and Miyazawa [35]. Our study showed that treatment
with CHCl3 plus CMN decreased cholesterol, triglycerides and LDL-
C, and increased HDL-C (Table 2). These results are in agreement
with Ruddick et al. [23], Inano et al. [36] and Nagata and Saito [37].
Schoonjans et al. [38] hypothesized that CMN is effective in lower-
ing TG by induction multiple of intra- and extracellular fatty acid
catabolism and utilization pathways.
We showed that curcumin acts as strong inhibitor to XO in
heart of group treated with CMN. Lin and Shih [39] reported
that curcumin can inhibit XO. In contrast, Pauff and Hille [40]
found that curcumin exhibit no inhibitory activity against XO. Also,
Shen and Ji [41] reported that curcumin bind weakly to XO while
its degradation products such as trans-6-(4-hydroxy-3-methoxy
phenyl)-2, 4-ioxo-5-hexenal, ferulic aldhdye, ferulic acid, feruloyl
methane and vanillin show inhibitory activities against XO. Besides,
we showed that administration of curcumin lead to decrease in
MPO enzyme activity in cardiac tissue. Kato et al. [42] showed
an inhibitory effect of curcumin on MPO-catalyzed tyrosylation
which attributed to quenching of the tryrosyl radical by poly phenol
and/or to an interaction between MPO and curcumin. Our results
showed that curcumin treatment returned the increased MDA lev-
els back to the control in co-treated group and decreasing in group
treated with CMN alone. This result is hand on hand with the
ndings of Chandran and Venugopal [43] who reported that admin-
istration of curcumin reserved the change induced by nicotine,
supporting the hypothesis that curcumin is effective antioxidative
agent. The present study demonstrated that total protein content
in heart did not change as compared with the control. These nd-
ings were in accordance with Nagata and Saito [37]. Treatment
with curcumin signicantly enhanced the total antioxidant capac-
ity, GSH and total thiol status in heart as compared with the control,
also attenuating the effect of CHCl3 in group treated with CMN
plus CHCl3 as compared with the control. Our results are compat-
ible with Khalil and Ali [44] and Kalpana and Menon [45]. Biswas
et al. [46] reported that curcumin has the ability to decrease sin-
glet oxygen, HO. and other anion radicals in cell free medium.
Another mechanism by which curcumin may prevent GSH deple-
tion, it could be mediated via increased biosynthesis of GSH. Up
regulation of GCS lead to increased GSH levels in the cell [47].
Also, treatment of rats with curcumin brings back the levels of
(CAT and GSH-Px) enzymes activities to near normal levels in group
treated with CMN plus CHCl3. Besides, GSH-Px and CAT enzymes
activities account signicant increase in cardiac tissue in group
treated with curcumin as compared with the control (Table 3). This
result is in accordance with Kalpana and Menon [45] and Masuda
et al. [48].
5. Conclusion
Our results reported that CHCl3 , capable of caused marked alter-
ation in some biochemical parameters, induced oxidative damage
and inhibited the activities of antioxidant enzymes. While, cur-
cumin administrated in combination with chloroform minimized
its hazardous. In addition, curcumin alone prove to be benecial
230 A.F. Salama et al. / Biomedicine & Preventive Nutrition 4 (2014) 225230
in decreasing the levels of free radicals and lipid, and increasing
antioxidant enzymes. Consequently, the exposure to chloroform
should be reduced and attention paid to disinfection by-products
of water chlorination. Furthermore, using diets rich in curcumin
could be benecial in alleviating chloroform toxicity.
Disclosure of interest
The authors have not supplied their declaration of conict of
interest.
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