Protective effect of Curcuma longa L. extract on CCl4-induced acute hepatic stress
Lee et al. BMC Res Notes
Protective effect of Curcuma longa L. extract on CCl -induced acute hepatic stress 4
Geum‑Hwa Lee 0
HwaY‑oung Lee 0
Min‑Kyung Choi 0
HanW‑ool Chung 0
HanJ‑ung Chae 0
0 Department of Pharmacology and New Drug Development Institute, Chonbuk National University Medical School , Jeonju, Chonbuk 561‐180 , Republic of Korea
Background: The Curcuma longa L. (CLL) rhizome has long been used to treat patients with hepatic dysfunction. CLL is a member of the ginger family of spices that are widely used in China, India, and Japan, and is a common spice, coloring, flavoring, and traditional medicine. This study was performed to evaluate the hepatoprotective activity of CLL extract and its active component curcumin in an acute carbon tetrachloride (CCl4)‑ induced liver stress model. Methods: Acute hepatic stress was induced by a single intraperitoneal injection of CCl4 (0.1 ml/kg body weight) in rats. CLL extract was administered once a day for 3 days at three dose levels (100, 200, and 300 mg/kg/day) and curcumin was administered once a day at the 200 mg/kg/day. We performed alanine transaminase (ALT) and aspartate transaminase (AST). activity analysis and also measured total lipid, triglyceride, and cholesterol levels, and lipid peroxidation. Results: At 100 g CLL, the curcuminoid components curcumin (901.63 ± 5.37 mg/100 g), bis‑ demethoxycurcumin (108.28 ± 2.89 mg/100 g), and demethoxycurcumin (234.85 ± 1.85 mg/100 g) were quantified through high liquid chromatography analysis. In CCl4‑ treated rats, serum AST and ALT levels increased 2.1‑ and 1.2‑ fold compared with the control. AST but not ALT elevation induced by CCl4 was significantly alleviated in CLL‑ and curcumin‑ treated rats. Peroxidation of membrane lipids in the liver was significantly prevented by CLL (100, 200, and 300 mg/kg/day) on tissue lipid peroxidation assay and immunostaining with anti‑ 4HNE antibody. We found that CLL extract and curcumin exhibited significant protection against liver injury by improving hepatic superoxide dismutase (p < 0.05) and glutathione peroxidase activity, and glutathione content in the CCl4‑ treated group (p < 0.05), leading to a reduced lipid peroxidase level. Conclusion: Our data suggested that CLL extract and curcumin protect the liver from acute CCl4‑ induced injury in a rodent model by suppressing hepatic oxidative stress. Therefore, CLL extract and curcumin are potential therapeutic antioxidant agents against acute hepatotoxicity.
Hepatotoxicity; CLL extract; Lipid peroxidation; GSH; Curcumin
Curcumin, the pure active component of Curcuma longa
L. (CLL) and the yellow pigment that is a characteristic
feature of curry , has been studied for its
anti-inflammatory , immunoregulatory , and other beneficial
effects in models of hepatic dysfunction. Researchers
have performed a variety of studies to develop natural
products that improve hepatic function, including studies
of CLL extracts. The mechanism of the protective effects
of CLL against hepatic dysfunction has been suggested
to be the inhibition of tumor necrosis factor
(TNF)induced apoptosis [4, 5]. As a polyphenolic antioxidant,
curcumin has been suggested to inhibit the activation of
fibrosis in vitro by reducing cell proliferation and
inducing apoptosis . The antioxidant effects of CLL extracts
and curcumin have also been studied in a rat model of
carbon tetrachloride (CCl4)-induced liver injury [6, 7].
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Similarly, the hepatoprotective activity of Silybum
marianum , Tridax procumbens , Andrographis
paniculata , and Eucommia ulmoides  have been
studied to develop herbal medicines and functional foods
to improve hepatic function. The systems used for
developing natural products and medicines are usually based
upon severe hepatic dysfunction models such as liver
necrosis, necrosis, and cirrhosis . However, people
are frequently exposed to subclinical hepatic stress, and,
if a hepatotoxin-associated hepatic event exceeds the
hepatic capacity for clearing these toxins, hepatic
function can be transiently decreased, leading to acute liver
failure. Chemicals often cause subclinical injury to the
liver that manifests only as abnormal liver enzyme levels,
such as AST. Drug-induced liver injury is responsible for
5% of all hospital admissions and 50% of all cases of acute
liver failure . Although treatment of acute liver
failure is considered clinically important [14, 15], few
preventive products or medicines have been established. In
this study, we evaluated the hepatoprotective activity of
CLL extract and curcumin in a CCl4-induced acute liver
toxicity rat model. Compared with other studies of
CCl4induced liver cirrhosis, fibrosis and other severe hepatic
toxicities [16–19], our research design is an
acute/transient toxicity study based on “a low dose of chemical
toxin and one time exposure without histological
abnormalities.” Considering that transient or acute toxicity can
be more frequently happening to human, the
acute/transient toxicity model might have a more importantly
clinical meaning than other chronic/severe toxicity. We also
sought to determine whether the antioxidant properties
of CLL extracts or curcumin are involved in the
protective effects against acute liver toxicity.
Korean CLL rhizomes were harvested from Jindo, Korea
and extracts were manufactured by the Ottogi
Corporation (Seoul, Korea). The roots were crushed and loaded
into an extractor. The first and second extractions were
carried out with 50% ethanol. The second extracts were
filtered and gathered with the first extracts and were
concentrated under reduced pressure (20 brix). Twenty
percent dextrin was added to the final extracts and they were
sterilized. The total yield was 18% based upon weight.
Quantitation of curcumin and curcuminoids using HPLC–
Curcumin and curcuminoids were prepared with 100%
methanol and quantitated with an HPLC system (Agilent
1100 series, Germany) equipped with a Zorbax Eclipse
C18 column (250 × 4.0 mm2). The mobile phase
consisted of 1% acetic acid in water (A) and 52% acetonitrile
(B); the column was equilibrated for 10 min in the mobile
phase and then washed with 100% (B) for 10 min. The
column was operated at room temperature with a 1 ml/
min flow rate. The injection volume was 5 μl, and
curcumin, curcuminoids, bis-demethoxycurcumin (BDMC),
and demethoxycurcumin (DMC) were detected at a
wavelength of 424 nm.
Reagents and chemicals
Carbon tetrachloride and curcumin were purchased from
Sigma Chemicals Co. (St. Louis, MO, USA). The
commercial kits used for assaying liver enzymes and
antioxidants are described below.
One hundred male Sprague–Dawley (SD) rats (250–
280 g) were purchased from Central Lab Animal Inc.
(Seoul, Korea) randomly assigned into groups. The
experimental animals were given free access to standard diet
and water in rooms maintained at 25 °C on 12-h light/
dark cycles. Four rats were sacrificed for preliminary
CCl4 toxicity testing, and 12 of the surviving rats were
placed in each of the following groups. Control rats were
injected with olive oil (vehicle; 0.1 ml/kg body weight,
i.p.) for 72 h. The curcumin group was intragastrically
administered with 200 mg/kg curcumin for 72 h. The CLL
group was intragastrically administered with 300 mg/
kg CLL extract. The CCl4 group received CCl4 (0.1 ml/
kg, i.p.) for 72 h. The CCl4-curcumin group received
curcumin (200 mg/kg) intragastrically before 3 days of CCl4
treatment. The CCl4-CLL groups received 100 mg/kg,
200 mg/kg, or 300 mg/kg of CLL extract intragastrically
before 3 days of CCl4 treatment. All of the experimental
protocols conducted on rats were performed in
accordance with internationally accepted principles for
laboratory animal use and care and approval was obtained from
the Care and Use of Laboratory Animals Committee of
Chonbuk National University Hospital. All procedures
were also approved by the Institutional Animal Care and
Use Committee of Chonbuk National University Hospital
(IACUC protocol CBU 150608-25).
Immunohistochemistry was performed using 4-μm-thick
deparaffinized liver tissue sections as described earlier
. Briefly, deparaffinized liver slices were incubated
overnight with antibodies against 4-hydroxynonenal
(4-HNE). Antibody detection was performed using the
DAKO EnVision + System Peroxidase/DAB kit.
AST and ALT activity
Serum levels of liver enzymes AST and ALT were
estimated using commercially available diagnostic kits (Cat.
AM101-K, Asan Pharm, Seoul, Korea) according to the
Lipid profile analysis
Total cholesterol, high-density lipoprotein (HDL)
cholesterol, low-density lipoprotein cholesterol (LDL), and
triglycerides (TG) were estimated using commercially
available diagnostic kits (Cat. AM203-K and AM 202-K,
The activity of SOD and glutathione peroxidase (GPx)
was analyzed using assay kits from Cayman according to
the manufacturer’s instructions (Cat.706002 and 703102,
Cayman, Ann Arbor, MI, USA).
The level of serum reduced and oxidized glutathione
(GSH and GSSG) was measured using a kit from
BioVision (Cat. K264, BioVision, Inc, CA, USA) according to
the manufacturer’s protocol.
Determination of tissue lipid peroxidation
The level of serum and liver lipid peroxidation was
measured using a thiobarbituric acid reactive substances
(TBARS) kit from Cayman (Cat. 10009055) according to
the manufacturer’s protocol.
Results are presented as the mean ± SEM. MicroCal
Origin software (Northampton, MA, USA) was used for all
statistical calculations. Differences were tested for
significance using one-way analysis of variance (ANOVA) with
Duncan’s multiple range test. Statistical significance was
set at p < 0.05.
Curcuminoid components of CLL turmeric
From the CLL extract, major compounds
including curcumin, bis-demethoxycurcumin (BDMC), and
demethoxycurcumin (DMC) (Fig. 1a) were
identified and quantified using HPLC. From 100-g CLL
extracts, curcumin (901.63 ± 5.37 mg/100 g), BDMC
(108.28 ± 2.89 mg/100 g), DMC (234.85 ± 1.85 mg/100 g)
and total curcuminoids (1244.76 ± 3.86 mg/100 g) were
quantified (Table 1). The representative HPLC
chromatogram is presented in Fig. 1b.
CLL turmeric extract and curcumin protect against the
CCl4‑induced toxicity profile
To examine the role of CLL turmeric extract and curcumin
in hepatic toxicity, the extract and curcumin were applied
to a CCl4-induced acute toxicity model. The serum activity
of AST was increased in the CCl4 group compared with
the control group (Fig. 2). CCl4-increased AST activity
was significantly reduced in the presence of CLL extract
or curcumin. Although serum ALT level showed a similar
pattern to AST level, there were no significance differences
between CCl4 and the combined CLL extract or curcumin
groups. Consistent with these data, AST serum level rather
than ALT has been used as a biochemical marker for early
acute hepatotoxicity [21, 22].
CLL turmeric extract does not affect serum lipid profile in a
CCl4‑induced acute toxicity model
Serum cholesterol, triglycerides, and LDL levels did not
vary significantly between the CCl4 group and the control
group. Administration of CLL extract alone resulted in a
non-significant change in lipid profiles compared to the
control group (Table 2).
CLL turmeric extract increases antioxidant enzymes in a
CCl4‑induced acute toxicity model
Liver activity of SOD and GPx was decreased in the CCl4
group compared with the control group (Fig. 3a, b). The
CCl4-induced decrease in SOD and GPx activity
recovered after treatment with CLL extract or curcumin. In
the SOD activity analysis, CLL extract showed a
dosedependent recovery effect; the highest CLL extract dose,
300 mg/kg, showed the greatest protective effect against
decreased SOD activity. In the GPx activity analysis, a
relatively low dose of 100 mg/kg CLL showed the greatest
recovery effect against decreased GPx activity, similar to
the recovery effect of curcumin.
CLL turmeric extract inhibits lipid peroxidation in a
CCl4‑induced acute toxicity model
Increased levels of reactive oxygen species (ROS) induce
membrane lipid peroxidation and the production of
associated by-products such as malondialdehyde (MDA) and
4HNE . As a product of lipid peroxidation, the MDA
level can reflect the liver lipid peroxidation level . In
damaged tissues, 4HNE has been found in higher
quantities during oxidative stress due to the increase in the
lipid peroxidation chain reaction . Figure 4a shows
that the liver MDA level increased significantly in
CCl4treated mice compared with the control group.
Treatment with CLL extract (200 and 300 mg/kg) reduced this
CCl4-induced increase in MDA. In addition, our results
showed that the number of 4-HNE-stained hepatocytes
increased in the CCl4-treated mice (Fig. 4b). Treatment
with CLL extract (100, 200, and 300 mg/kg) reduced the
CCl4-induced increase in 4-HNE-stained cells. Curcumin
treatment consistently showed similar effects to CLL
extract. These data collectively suggest that CLL extract
protects the liver against CCl4-induced oxidative stress.
Fig. 1 HPLC analysis of CLL extract. a Chemical structure of curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC). b HPLC
chromatogram analysis of CLL extract
Table 1 Analysis of CLL extract with HPLC
Curcuminoids BDMC DMC
The values are shown as mean ± SEM (n = 3)
CLL turmeric extract and curcumin affect GSH profiles in a
CCl4‑induced acute toxicity model
GSH is an important cellular antioxidant that protects
cells against ROS-induced liver injury . The efficient
transformation of GSH to GSSG has been suggested to be
a marker of redox capacity to explain the cellular redox
environment [27, 28]. The data shown in Fig. 5a confirm
the decrease in GSH level in CCl4-treated rats. CLL extract
dose-dependently rescued the CCl4-induced decrease in
liver GSH level. The oxidation product, GSSG, showed a
similar pattern (data not shown). Compared to the control
group, the level of total glutathione, including GSSG, was
significantly reduced by CCl4 (Fig. 5b). CLL extract
significantly recovered the decreased total GSH level in a
dosedependent manner. The recovery effect of 300 mg/kg CLL
was similar to that of curcumin, suggesting that CLL extract
and its pure component curcumin protect against oxidative
stress by enhancing the intrahepatic redox capacity.
Fig. 2 Effects of CLL extract on serum liver biomarkers. Serum AST (a) and ALT (b) were analyzed in control, 0.1 ml/kg CCl4, CCl4 with curcumin, CCl4
with CLL extract (100, 200, or 300 mg/kg), 200 mg/kg curcumin alone, and 300 mg/kg CLL extract alone groups. Each bar represents the mean value
of experiments performed in triplicate ± S.E.M. (n = 10). *p < 0.05 compared with the CCl4 group
Table 2 Effect of CLL extract on TG, total-cholesterol, LDL-cholesterol, and HDL-cholesterol at CCl4-induced acute stress
Serum levels (mg/dl)
In this report, we showed that the application of CLL
extracts markedly inhibited acute hepatic failure in a
rat model of acute hepatic injury, which was induced at
least in part by free radical formation. Our data suggest
that CLL and its active component curcumin are
effective against acute liver stress by enhancing redox capacity
and antioxidant enzyme activity. In this study, the acute
CCl4 toxicity model was used to evaluate the efficacy of
CLL extract in acute hepatic stress. Acute CCl4
administration is a widely used experimental model that mimics
the acute liver failure caused by toxic substances [29–31].
Although the liver plays a key role in transforming and
clearing chemicals, certain medicinal agents, when taken
in excess, may injure the organ. Other chemical agents or
industrial agents can also induce hepatotoxicity. These
chemicals often cause subclinical injury to the liver,
increasing liver enzymes, but not causing pathological
abnormalities such as histological and hepatosteatosis
status changes. Drug-induced acute liver failure is an
important clinical issue , and thus therapeutic and
preventive strategies against acute liver toxicity need to
be developed. In the acute toxicity model, CCl4 did not
induce hepatic lipid accumulation (Table 2), but did
increase activity of the main liver enzyme AST (Fig. 2),
a routinely observed acute toxicity-associated clinical
symptom. CLL extract inhibited this increase in AST
activity. Although AST and ALT aminotransferases are
both highly concentrated in the liver, this model showed
an increase in AST, but not ALT. A similar pattern was
reported recently in cases of acute hepatic and transient
stress [22, 32]. AST is localized in the mitochondria,
100 200 300
Fig. 3 Effects of CLL extract on SOD and GPx activity. a Hepatic SOD
and b GPx activity was analyzed in control, 0.1 ml/kg CCl4, CCl4 with
curcumin, CCl4 with CLL extract (100, 200, or 300 mg/kg), 200 mg/
kg curcumin alone, and 300 mg/kg CLL extract alone groups. Each
bar represents the mean value of experiments performed in tripli‑
cate ± S.E.M. (n = 10). *p < 0.05 compared with the CCl4 group
whereas ALT is distributed throughout the cytoplasm.
In the case of hepatic stress, mitochondrial damage with
ROS accumulation tends to increase the level of AST
rather than ALT [33, 34]. The manifestation of acute
hepatotoxicity in acute stress is highly variable, ranging
from asymptomatic elevation of liver enzymes to
fulminant hepatic failure . Because of the considerable
disease burden, there is growing interest in
understanding acute hepatic failure. One representative system for
understanding acute hepatic failure is the CCl4 model.
Fig. 4 Effects of CLL extract on hepatic lipid peroxidation. The
formation of a malondialdehyde (MDA) and b 4‑HNE was analyzed in
control, 0.1 ml/kg CCl4, CCl4 with 200 mg/kg curcumin, CCl4 with CLL
extract (100, 200, or 300 mg/kg), 200 mg/kg curcumin, and 300 mg/
kg CLL extract groups. Each bar represents the mean value of experi‑
ments performed in triplicate ± S.E.M. (n = 10). *p < 0.05 compared
with the CCl4 group
Fig. 5 Effects of CLL extract on hepatic redox capacity. a Reduced glutathione (GSH), b total GSH, c GSH/oxidized glutathione (GSSG) ratio and
d GSH+GSSG content were analyzed in control, 0.1 ml/kg CCl4, CCl4 with 200 mg/kg curcumin, CCl4 with CLL extract (100, 200, or 300 mg/kg),
200 mg/kg curcumin, and 300 mg/kg CLL extract groups. Each bar represents the mean value of experiments performed in triplicate ± S.E.M.
(n = 10). *p < 0.05 compared with the CCl4 group
Metabolic activation of CCl4 by mixed function oxidases
is required to induce hepatotoxicity. The initial step in
liver injury induction by CCl4 is mainly its
dehalogenation by cytochrome p-450 2E1 (CYP2E1) to a
methyl free radical (CCl3 ), which leads to hepatic toxicity
. A single injection of CCl4 did not induce hepatic
lipid accumulation, but simply resulted in increased AST
activity and hepatic ROS accumulation. The damage
pattern was transient, with recovery to a normal state (data
not shown). The main point of this study was transient
stress. Hepatic stress that occurs during life is typically
transient; at rest, the liver recovers after a short period
of time and therefore the stress often goes unnoticed
clinically. The initial hypothesis of this study was that
foods that prevent routine acute stress might be good for
maintaining hepatic health; in this regard, CLL extract
was a strong candidate for testing the hypothesis.
Curcumin and CLL extracts have been frequently studied
with regard to hepatoprotective function. However, the
main design of those studies was based on a high level
of toxin-induced hepatic dysfunction [2, 37, 38], which
varied from the present study’s design; acute or
transient toxicity without abnormal liver function except
liver enzyme. Throughout this study, CLL extract and its
active component curcumin showed antioxidant enzyme
activity and a regulatory effect against the accumulation
of ROS, including lipid peroxidation, resulting in a
protective effect against CCl4-induced acute hepatotoxicity.
Although the suggested antioxidant mechanism is
similar to its application in severe hepatotoxicity, this study
strongly suggests the possible application of CLL to
transient or acute hepatic stress conditions. Other studies
indicated that CLL treatment induces an augmentation
of hepatic Nrf-2 protein levels  and stimulates
antioxidant activity [38–40]. Curcumin, the main component
of CLL, significantly reduced the CCl4-induced increase
in hepatic MDA , implying that CLL exerts protective
effects against CCl4-induced liver damage by
preventing lipid peroxide formation and by blocking the
oxidative chain reaction . Our results indicated that CCl4
administration led to a marked depletion of glutathione
(GSH) level in the liver. GSH, a cytosolic tripeptide, is
ubiquitously present in all cell types at millimolar
concentrations and is the major non-enzymatic regulator of
intracellular redox homeostasis . GSH is oxidized to
GSSG by the enzymatic reaction catalyzed by glutathione
peroxidase (GPx), which is reduced back to GSH by
glutathione reductase . Our results showed a drastic
reduction in the activity of GPx in the liver caused by
CCl4, which could compromise the defenses of the liver.
Our results suggested that CCl4 administration decreased
the concentrations of reduced GSH and total GSH in
the liver, altering the redox status of the cells, and that
treatment with CLL extract and its active component
curcumin led to recovery of redox balance. Increases
in the levels of reduced GSH and total GSH upon
treatment with CLL extracts may be involved in the protective
mechanism against CCl4-induced liver toxicity.
In this study, CLL extract and its active component
curcumin showed protective activity against CCl4-induced
hepatotoxicity. In addition to antioxidant activity, CLL
extracts and their active components might play a role in
restoring the liver redox capacity, as represented by
GSHGSSG cycling capacity. In future studies, derivatives of
CLL extracts should be examined in various
experimental models of acute toxicity in addition to CCl4. Active
derivatives may be potential drug candidates for acute
liver failure and toxicity.
CLL: Curcuma longa L. turmeric; CCl4: carbon tetrachloride; AST: aspartate
transaminase; ALT: alanine transaminase; MDA: malondialdehyde; SOD: super‑
oxide dismutase; GPx: glutathione peroxidase; 4‑HNE: 4‑hydroxynonenal; ROS:
reactive oxygen species; CYP2E1: cytochrome p‑450 2E1; BDMC: demethoxy‑
curcumin; DMC: demethoxycurcumin; LDL: low‑ density lipoprotein; HDL: high
GHL, HYL, HWC, and MKC made substantial contributions to the conception
and design of the study, acquisition of data, and analysis and interpreta‑
tion of data. SWK was involved in the acquisition of data and participated in
designing study. HJC was involved in the conception and design of the study,
and revised the manuscript critically for important intellectual content. All
authors read and approved the final manuscript.
This work was supported by the “Food Functionality Evaluation Program”
under the Ministry of Agriculture, Food, and Rural Affairs and in part by the
Korea Food Research Institute.
The authors declare that they have no competing interests.
Availability of data and materials
The datasets supporting the conclusions of this article are included within the
article and all datasets supporting our findings are available. A sample of the
CLL used in this study has been deposited at the Ottogi company.
Ethics approval and consent to participate
All procedures in the animal studies complied with standards for the care and
use of experimental animals and were approved by Animal Care Committee of
Chonbuk National University Hospital (IACUC protocol CBU 150608‑25).
1. Rezaei‑Moghadam A , Mohajeri D , Rafiei B , Dizaji R , Azhdari A , Yegane ‑ hzad M , Shahidi M , Mazani M. Effect of turmeric and carrot seed extracts on serum liver biomarkers and hepatic lipid peroxidation, antioxidant enzymes and total antioxidant status in rats . Bioimpacts . 2012 ; 2 ( 3 ): 151 - 7 .
2. Lee HS , Li L , Kim HK , Bilehal D , Li W , Lee DS , Kim YH . The protective effects of Curcuma longa Linn. extract on carbon tetrachloride‑induced hepatotoxicity in rats via upregulation of Nrf2 . J Microbiol Biotechnol . 2010 ; 20 ( 9 ): 1331 - 8 .
3. Nagpal M , Sood S. Role of curcumin in systemic and oral health: an overview . J Nat Sci Biol Med . 2013 ; 4 ( 1 ): 3 - 7 .
4. Li M , Wu Z , Niu W , Wan Y , Zhang L , Shi G , Xi X. The protective effect of curcumin against the 19 kDa Mycobacterium tuberculosis protein‑induced inflammation and apoptosis in human macrophages . Mol Med Rep . 2014 ; 10 ( 6 ): 3261 - 7 .
5. Xu J , Fu Y , Chen A. Activation of peroxisome proliferator‑activated receptor‑ gamma contributes to the inhibitory effects of curcumin on rat hepatic stellate cell growth . Am J Physiol Gastrointest Liver Physiol . 2003 ; 285 ( 1 ): G20 - 30 .
6. Wu SJ , Lin YH , Chu CC , Tsai YH , Chao JC . Curcumin or saikosaponin a improves hepatic antioxidant capacity and protects against CCl4‑induced liver injury in rats . J Med Food . 2008 ; 11 ( 2 ): 224 - 9 .
7. Fu Y , Zheng S , Lin J , Ryerse J , Chen A. Curcumin protects the rat liver from CCl4‑ caused injury and fibrogenesis by attenuating oxidative stress and suppressing inflammation . Mol Pharmacol . 2008 ; 73 ( 2 ): 399 - 409 .
8. Shahbazi F , Sadighi S , Dashti‑Khavidaki S , Shahi F , Mirzania M , Abdollahi A , Ghahremani MH . Effect of silymarin administration on cisplatin nephrotoxicity: report from a pilot, randomized, double‑blinded, placebo ‑ controlled clinical trial . Phytother Res . 2015 ; 29 ( 7 ): 1046 - 53 .
9. Ravikumar V , Shivashangari KS , Devaki T. Effect of Tridax procumbens on liver antioxidant defense system during lipopolysaccharideinduced hepatitis in d‑ galactosamine sensitised rats . Mol Cell Biochem . 2005 ; 269 ( 1-2 ): 131 - 6 .
10. Zou Y , Xiong H , Xiong H , Lu T , Zhu F , Luo Z , Yuan X , Wang Y. A polysaccharide from Andrographis paniculata induces mitochondrial‑mediated apoptosis in human hepatoma cell line (HepG2) . Tumour Biol . 2015 ; 36 ( 7 ): 5179 - 86 .
11. Jin CF , Li B , Lin SM , Yadav RK , Kim HR , Chae HJ . Mechanism of the inhibitory effects of Eucommia ulmoides Oliv. cortex extracts (EUCE) in the CCl 4‑induced acute liver lipid accumulation in rats . Int J Endocrinol . 2013 ; 2013 : 751854 .
12. Wang K. Molecular mechanisms of hepatic apoptosis . Cell Death Dis . 2014 ; 5 : e996 .
13. Ostapowicz G , Fontana RJ , Schiodt FV , Larson A , Davern TJ , Han SH , McCashland TM , Shakil AO , Hay JE , Hynan L , et al. Results of a prospective study of acute liver failure at 17 tertiary care centers in the United States . Ann Intern Med . 2002 ; 137 ( 12 ): 947 - 54 .
14. Jalan R. Acute liver failure: current management and future prospects . J Hepatol . 2005 ; 42 (Suppl (1)): S115 - 23 .
15. Polson J , Lee WM , American Association for the Study of Liver D. AASLD position paper: the management of acute liver failure . Hepatology . 2005 ; 41 ( 5 ): 1179 - 97 .
16. Domitrovic R , Jakovac H , Tomac J , Sain I. Liver fibrosis in mice induced by carbon tetrachloride and its reversion by luteolin . Toxicol Appl Pharmacol . 2009 ; 241 ( 3 ): 311 - 21 .
17. Louka ML , Ramzy MM . Involvement of fibroblast‑specific protein 1 (S100A4) and matrix metalloproteinase‑13 (MMP ‑13) in CCl4‑induced reversible liver fibrosis . Gene . 2016 ; 579 ( 1 ): 29 - 33 .
18. Munoz‑ Ortega MH , Llamas‑Ramirez RW , Romero ‑Delgadillo NI , EliasFlores TG , Tavares‑Rodriguez Ede J , Campos‑Esparza Mdel R , CervantesGarcia D , Munoz‑Fernandez L , Gerardo ‑Rodriguez M , Ventura‑ Juarez J. Doxazosin treatment attenuates carbon tetrachloride‑induced liver fibrosis in hamsters through a decrease in transforming growth factor beta secretion . Gut Liver . 2016 ; 10 ( 1 ): 101 - 8 .
19. Wang Z , Zhang Z , Du N , Wang K , Li L. Hepatoprotective effects of grape seed procyanidin B2 in rats with carbon tetrachloride‑induced hepatic fibrosis . Altern Ther Health Med . 2015 ; 21 (Suppl 2): 12 - 21 .
20. Domitrovic R , Jakovac H , Marchesi VV , Sain I , Romic Z , Rahelic D. Preventive and therapeutic effects of oleuropein against carbon tetrachlorideinduced liver damage in mice . Pharmacol Res . 2012 ; 65 ( 4 ): 451 - 64 .
21. Tossige‑ Gomes R , Ottone VO , Oliveira PN , Viana DJ , Araujo TL , Gripp FJ , Rocha‑ Vieira E. Leukocytosis , muscle damage and increased lymphocyte proliferative response after an adventure sprint race . Braz J Med Biol Res . 2014 ; 47 ( 6 ): 492 - 8 .
22. Andres‑Hernando A , Altmann C , Bhargava R , Okamura K , Bacalja J , Hunter B , Ahuja N , Soranno D , Faubel S. Prolonged acute kidney injury exacerbates lung inflammation at 7 days post‑acute kidney injury . Physiol Rep . 2014 ; 2 ( 7 ): e12084 .
23. Singh R , Wang Y , Schattenberg JM , Xiang Y , Czaja MJ . Chronic oxidative stress sensitizes hepatocytes to death from 4‑hydroxynonenal by JNK/c‑ Jun overactivation . Am J Physiol Gastrointest Liver Physiol . 2009 ; 297 ( 5 ): G907 - 17 .
24. Misra HP , Rabideau C. Pirfenidone inhibits NADPH‑ dependent microsomal lipid peroxidation and scavenges hydroxyl radicals . Mol Cell Biochem . 2000 ; 204 ( 1-2 ): 119 - 26 .
25. Hartley DP , Kolaja KL , Reichard J , Petersen DR. 4 ‑ Hydroxynonenal and malondialdehyde hepatic protein adducts in rats treated with carbon tetrachloride: immunochemical detection and lobular localization . Toxicol Appl Pharmacol . 1999 ; 161 ( 1 ): 23 - 33 .
26. Srivastava S , Sinha D , Saha PP , Marthala H, D'Silva P. Magmas functions as a ROS regulator and provides cytoprotection against oxidative stressmediated damages . Cell Death Dis . 2014 ; 5 : e1394 .
27. Nur E , Verwijs M , de Waart DR , Schnog JJ , Otten HM , Brandjes DP , Biemond BJ , Elferink RP , Group CS. Increased efflux of oxidized glutathione (GSSG) causes glutathione depletion and potentially diminishes antioxidant defense in sickle erythrocytes . Biochim Biophys Acta . 2011 ; 1812 (11): 1412 - 7 .
28. Mizuashi M , Ohtani T , Nakagawa S , Aiba S. Redox imbalance induced by contact sensitizers triggers the maturation of dendritic cells . J Invest Dermatol . 2005 ; 124 ( 3 ): 579 - 86 .
29. Ma JQ , Li Z , Xie WR , Liu CM , Liu SS . Quercetin protects mouse liver against CCl(4)‑induced inflammation by the TLR2/4 and MAPK/NF‑kappaB pathway . Int Immunopharmacol . 2015 ; 28 ( 1 ): 531 - 9 .
30. Cao G , Li Q , Chen X , Cai H , Tu S. Hepatoprotective effect of superfine particles of herbal medicine against CCl4‑induced acute liver damage in rats . Biomed Res Int . 2014 ; 2014 : 934732 .
31. Li W , Wu Y , Zhu C , Wang Z , Gao R , Wu Q. Anti‑fibrosis effects of Huisheng oral solution in CCl4‑induced hepatic fibrosis in rat . Indian J Pharmacol . 2014 ; 46 ( 2 ): 216 - 21 .
32. Wang XY , Luo JP , Chen R , Zha XQ , Wang H. The effects of daily supplementation of Dendrobium huoshanense polysaccharide on ethanolinduced subacute liver injury in mice by proteomic analysis . Food Function . 2014 ; 5 ( 9 ): 2020 - 35 .
33. Shen J , Zhang J , Wen J , Ming Q , Zhang J , Xu Y. Correlation of serum alanine aminotransferase and aspartate aminotransferase with coronary heart disease . Int J Clin Exp Med . 2015 ; 8 ( 3 ): 4399 - 404 .
34. Hu Z , Lausted C , Yoo H , Yan X , Brightman A , Chen J , Wang W , Bu X , Hood L. Quantitative liver‑specific protein fingerprint in blood: a signature for hepatotoxicity . Theranostics . 2014 ; 4 ( 2 ): 215 - 28 .
35. Jiao J , Friedman SL , Aloman C. Hepatic fibrosis . Curr Opin Gastroenterol . 2009 ; 25 ( 3 ): 223 - 9 .
36. Khan RA , Khan MR , Sahreen S. CCl4‑induced hepatotoxicity: protective effect of rutin on p53, CYP2E1 and the antioxidative status in rat . BMC Complement Altern Med . 2012 ; 12 : 178 .
37. Park EJ , Jeon CH , Ko G , Kim J , Sohn DH . Protective effect of curcumin in rat liver injury induced by carbon tetrachloride . J Pharm Pharmacol . 2000 ; 52 ( 4 ): 437 - 40 .
38. Deshpande UR , Gadre SG , Raste AS , Pillai D , Bhide SV , Samuel AM . Protective effect of turmeric (Curcuma longa L.) extract on carbon tetrachlorideinduced liver damage in rats . Indian J Exp Biol . 1998 ; 36 ( 6 ): 573 - 7 .
39. Sharma OP . Antioxidant activity of curcumin and related compounds . Biochem Pharmacol . 1976 ; 25 ( 15 ): 1811 - 2 .
40. Salama SM , Abdulla MA , AlRashdi AS , Ismail S , Alkiyumi SS , Golbabapour S. Hepatoprotective effect of ethanolic extract of Curcuma longa on thioacetamide induced liver cirrhosis in rats . BMC Complement Altern Med . 2013 ; 13 : 56 .
41. Kim Y , You Y , Yoon HG , Lee YH , Kim K , Lee J , Kim MS , Kim JC , Jun W. Hepatoprotective effects of fermented Curcuma longa L. on carbon tetrachloride‑induced oxidative stress in rats . Food Chem . 2014 ; 151 : 148 - 53 .
42. Circu ML , Aw TY . Glutathione and modulation of cell apoptosis . Biochim Biophys Acta . 2012 ; 1823 (10): 1767 - 77 .
43. Franco R , Cidlowski JA . Apoptosis and glutathione: beyond an antioxidant . Cell Death Differ . 2009 ; 16 ( 10 ): 1303 - 14 .