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1.  Hepatoprotection with a chloroform extract of Launaea procumbens against CCl4-induced injuries in rats 
Background
Launaea procumbens (Asteraceae) is used as a folk medicine to treat hepatic disorders in Pakistan. The effect of a chloroform extract of Launaea procumbens (LPCE) was evaluated against carbon-tetrachloride (CCl4)-induced liver damage in rats.
Methods
To evaluate the hepatoprotective effects of LPCE, 36 male Sprague–Dawley rats were equally divided into six groups. Animals of group 1 (control) had free access to food and water. Group II received 3 ml/kg of CCl4 (30% in olive oil v/v) via the intraperitoneal route twice a week for 4 weeks. Group III received 1 ml of silymarin via gavage (100 mg/kg b.w.) after 48 h of CCl4 treatment whereas groups IV and V were given 1 ml of LPCE (100 and 200 mg/kg b.w., respectively) after 48 h of CCl4 treatment. Group VI received 1 ml of LPCE (200 mg/kg b.w.) twice a week for 4 weeks. The activities of the antioxidant enzymes catalase, peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), glutathione reductase (GSR), glutathione (GSH) and lipid peroxidation (thiobarbituric acid reactive substances (TBARS)) were measured in liver homogenates. DNA damage, argyrophilic nucleolar organizer regions (AgNORs) counts and histopathology were studied in liver samples. Serum was analyzed for various biochemical parameters. Phytochemical composition in LPCE was determined through high-performance liquid chromatography (HPLC).
Results
LPCE inhibited lipid peroxidation, and reduced the activities of aspartate transaminase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase in serum induced by CCl4. GSH contents were increased as were the activities of antioxidant enzymes (catalase, SOD, GST, GSR, GSH-Px) when altered due to CCl4 hepatotoxicity. Similarly, absolute liver weight, relative liver weight and the number of hepatic lesions were reduced with co-administration of LPCE. Phyochemical analyses of LPCE indicated that it contained catechin, kaempferol, rutin, hyperoside and myricetin.
Conclusion
These results indicated that Launaea procumbens efficiently protected against the hepatotoxicity induced by CCl4 in rats, possibly through the antioxidant effects of flavonoids present in LPCE.
doi:10.1186/1472-6882-12-114
PMCID: PMC3492157  PMID: 22862950
Launaea procumbens; Hepatic injuries; Flavonoids; Antioxidant enzymes; Carbon tetrachloride
2.  Protective effects of Sonchus asper (L.) Hill, (Asteraceae) against CCl4-induced oxidative stress in the thyroid tissue of rats 
Background
Sonchus asper (L.) Hill, (Asteraceae) is used in Pakistan as a traditional (“folk”) medicine for the treatment of hormonal disorders and oxidative stress. The present study was aimed to evaluate the efficacy of Sonchus asper (L.) Hill, (Asteraceae) methanolic extract (SAME) on hormonal dysfunction in thyroid tissue after carbon tetrachloride (CCl4)-induced oxidative stress.
Methods
To examine the effects of SAME against the oxidative stress of CCl4 in thyroid tissue, 30 male albino rats were used. Protective effects of SAME were observed on thyroid hormonal levels, activities of antioxidant enzymes, lipid peroxidation (TBARS) and DNA damage.
Results
Treatment with CCl4 significantly (P<0.01) reduced the levels of T3 and T4 and increased TSH levels. CCl4 exposure in rats reduced the activities of antioxidant enzymes but increased lipid peroxidation and DNA damage. Co-administration of SAME significantly (P<0.01) improved these alterations with respect to hormonal levels, activities of antioxidant enzymes and lipid peroxidation close to those seen in control rats.
Conclusion
These results suggest that SAME can protect thyroid tissue against oxidative damage, possibly through the antioxidant effects of its bioactive compounds.
doi:10.1186/1472-6882-12-181
PMCID: PMC3485175  PMID: 23043630
Sonchus asper; Oxidative stress; Antioxidant enzymes; Carbon tetrachloride; T3; T4
3.  CCl4-induced hepatotoxicity: protective effect of rutin on p53, CYP2E1 and the antioxidative status in rat 
Background
Rutin is a polyphenolic natural flavonoid which possesses antioxidant and anticancer activity. In the present study the hepatoprotective effect of rutin was evaluated against carbon tetrachloride (CCl4)-induced liver injuries in rats.
Methods and materials
24 Sprague–Dawley male rats were equally divided into 4 groups for the assessment of hepatoprotective potential of rutin. Rats of group I (control) received only vehicles; 1 ml/kg bw of saline (0.85%) and olive oil (3 ml/kg) and had free access to food and water. Rats of group II, III and IV were treated with CCl4 (30% in olive oil, 3 ml/kg bw) via the intraperitoneal route twice a week for four weeks. The rutin at the doses of 50 and 70 mg/kg were administered intragastrically after 48 h of CCl4 treatment to group III and IV, respectively. Protective effect of rutin on serum enzyme level, lipid profile, activities of antioxidant enzymes and molecular markers were calculated in CCl4-induced hepatotoxicity in rat.
Results
Rutin showed significant protection with the depletion of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (γ-GT) in serum as was raised by the induction of CCl4. Concentration of serum triglycerides, total cholesterol and low density lipoproteins was increased while high-density lipoprotein was decreased with rutin in a dose dependent manner. Activity level of endogenous liver antioxidant enzymes; catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSHpx), glutathione-S-transferase (GST) and glutathione reductase (GSR) and glutathione (GSH) contents were increased while lipid peroxidation (TBARS) was decreased dose dependently with rutin. Moreover, increase in DNA fragmentation and oxo8dG damages while decrease in p53 and CYP 2E1 expression induced with CCl4 was restored with the treatment of rutin.
Conclusion
From these results, it is suggested that rutin possesses hepatoprotective properties.
doi:10.1186/1472-6882-12-178
PMCID: PMC3519517  PMID: 23043521
Hepatotoxicity; Rutin; p53; CYP 2E1; Antioxidant enzymes
4.  Prevention of hepatorenal toxicity with Sonchus asper in gentamicin treated rats 
Background
Sonchus asper possesses antioxidant capacity and is used in liver and kidney disorders. We have investigated the preventive effect of methanolic extract of Sonchus asper (SAME) on the gentamicin induced alterations in biochemical and morphological parameters in liver and kidneys of Sprague-Dawley male rat.
Methods
Acute oral toxicity studies were performed for selecting the therapeutic dose of SAME. 30 Sprague-Dawley male rats were equally divided into five groups with 06 animals in each. Group I received saline (0.5 ml/kg bw; 0.9% NaCl) while Group II administered with gentamicin 0.5 ml (100 mg/kg bw; i.p.) for ten days. Animals of Group III and Group IV received gentamicin and SAME 0.5 ml at a dose of 100 mg/kg bw and 200 mg/kg bw, respectively while Group V received only SAME at a dose of 200 mg/kg bw. Biochemical parameters including aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), γ-glutamyltransferase (γ-GT), total cholesterol, triglycerides, total protein, albumin, creatinine, blood urea nitrogen (BUN), total bilirubin and direct bilirubin were determined in serum collected from various groups. Urinary out puts were measured in each group and also assessed for the level of protein and glucose. Lipid peroxides (TBARS), glutathione (GSH), DNA injuries and activities of antioxidant enzymes; catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) were determined in liver and renal samples. Histopathological studies of liver and kidneys were also carried out.
Results
On the basis of acute oral toxicity studies, 2000 mg/kg bw did not induce any toxicity in rats, 1/10th of the dose was selected for preventive treatment. Gentamicin increased the level of serum biomarkers; AST, ALT, ALP, LDH, γ-GT, total cholesterol, triglycerides, total protein, albumin, creatinine, BUN, total and direct bilirubin; as were the urinary level of protein, glucose, and urinary output. Lipid peroxidation (TBARS) and DNA injuries increased while GSH contents and activities of antioxidant enzymes; CAT, POD, SOD decreased with gentamicin in liver and kidney samples. SAME administration, dose dependently, prevented the alteration in biochemical parameters and were supported by low level of tubular and glomerular injuries induced with gentamicin.
Conclusion
These results suggested the preventive role of SAME for gentamicin induced toxicity that could be attributed by phytochemicals having antioxidant and free radical scavenging properties.
doi:10.1186/1472-6882-11-113
PMCID: PMC3305921  PMID: 22082144
5.  Protective effect of Launaea procumbens (L.) on lungs against CCl4-induced pulmonary damages in rat 
Background
Launaea procumbens (L.) is traditionally used in the treatment of various human ailments including pulmonary damages. The present study was arranged to evaluate the role of Launaea procumbens methanol extract (LME) against carbon tetrachloride (CCl4) induced oxidative pulmonary damages in rat.
Methods
36 Sprague–Dawley male rats (170-180 g) were randomly divided into 06 groups. After a week of acclamization, group I was remained untreated while group II was given olive oil intraperitoneally (i.p.) and dimethyl sulfoxide (DMSO) orally, groups III, IV, V and VI were administered CCl4, 3 ml/kg body weight (30% in olive oil i.p.). Groups IV, V were treated with 100 mg/kg, 200 mg/kg of LME whereas group VI was administered with 50 mg/kg body weight of rutin (RT) after 48 h of CCl4 treatment for four weeks. Antioxidant profile in lungs were evaluated by estimating the activities of antioxidant enzymes; catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione-S-transferase (GST), glutathione reductase (GSR), glutathione peroxidase (GSH-Px), quinone reductase (QR) and reduced glutathione (GSH). CCl4-induced lipid peroxidation was determined by measuring the level of thiobarbituric acid reactive substances (TBARS) with conjugation of deoxyribonucleic acid (DNA) damages, argyrophilic nucleolar organizer regions (AgNORs) counts and histopathology.
Results
Administration of CCl4 for 6 weeks significantly (p < 0.01) reduced the activities of antioxidant enzymes and GSH concentration while increased TBARS contents and DNA damages in lung samples. Co-treatment of LME and rutin restored the activities of antioxidant enzymes and GSH contents. Changes in TBARS concentration and DNA fragmentation were significantly (p < 0.01) decreased with the treatment of LME and rutin in lung. Changes induced with CCl4 in histopathology of lungs were significantly reduced with co-treatment of LME and rutin.
Conclusion
Results of present study revealed that LME could protect the lung tissues against CCl4-induced oxidative stress possibly by improving the antioxidant defence system.
doi:10.1186/1472-6882-12-133
PMCID: PMC3522065  PMID: 22909101
Launaea procumbens; Lungs; CCl4; Antioxidant enzymes; TBARS; GSH
6.  Brain antioxidant markers, cognitive performance and acetylcholinesterase activity of rats: efficiency of Sonchus asper 
Background
Sonchus asper (SA) is traditionally used as a folk medicine to treat mental disorders in Pakistan. The aim of this study was to investigate the effect of polyphenolic rich methanolic fraction of SA on cognitive performance, brain antioxidant activities and acetylcholinesterase activity in male rats.
Methods
30 male Sprague–Dawley rats were equally divided into three groups in this study. Animals of group I (control) received saline (vehicle), group II received SA (50 mg/kg) body weight (b.w.), and group III treated with SA (100 mg/kg b.w.,) orally in dimethyl sulphoxide (DMSO) for 7 days. The effect of SA was checked on rat cognitive performance, brain antioxidatant and acetylcholinesterase activities. Evaluation of learning and memory was assessed by a step-through a passive avoidance test on day 6 after two habituation trials and an initial acquisition trial on day 5. Antioxidant potential was determined by measuring activities of superoxide dismutase (SOD), catalase (CAT), contents of thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH) in whole-brain homogenates. Acetylcholinesterase (AChE) activity was determined by the colorimetric method.
Results
Results showed that 100 mg/kg b.w., SA treated rats exhibited a significant improvement in learning and memory (step-through latency time). SA administration reduced lipid peroxidation products and elevated glutathione levels in the SA100-treated group. Furthermore, salt and detergent soluble AChE activity was significantly decreased in both SA-treated groups. Short-term orally supplementation of SA showed significant cognitive enhancement as well as elevated brain antioxidant enzymes and inhibited AChE activity.
Conclusion
These findings stress the critical impact of Sonchus asper bioactive components on brain function.
doi:10.1186/1744-9081-8-21
PMCID: PMC3527136  PMID: 22591917
Sonchus asper; Cognitive performance; Acetylcholinesterase activity; Antioxidant enzymes
7.  Hispolon Protects against Acute Liver Damage in the Rat by Inhibiting Lipid Peroxidation, Proinflammatory Cytokine, and Oxidative Stress and Downregulating the Expressions of iNOS, COX-2, and MMP-9 
The hepatoprotective potential of hispolon against carbon tetrachloride (CCl4)-induced liver damage was evaluated in preventive models in rats. Male rats were intraperitoneally treated with hispolon or silymarin once daily for 7 consecutive days. One hour after the final hispolon or silymarin treatment, the rats were injected with CCl4. Administration with hispolon or silymarin significantly decreased the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in serum and increased the activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and glutathione (GSH) content and decreased the malondialdehyde (MDA) content in liver compared with CCl4-treated group. Liver histopathology also showed that hispolon reduced the incidence of liver lesions induced by CCl4. In addition, hispolon decreased nitric oxide (NO) production and tumor necrosis factor (TNF-α), inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) activation in CCl4-treated rats. We also examined the involvement of matrix metalloproteinase (MMP)-9 in the development of CCl4-induced liver damage in rats. Hispolon inhibited the expression of MMP-9 protein, indicating that MMP-9 played an important role in the development of CCl4-induced rat liver damage. Therefore, we speculate that hispolon protects rats from liver damage through their prophylactic redox balancing ability and anti-inflammation capacity.
doi:10.1155/2012/480714
PMCID: PMC3195309  PMID: 22013489
8.  Evaluation of hepatoprotective activity of Cleome viscosa Linn. extract 
Indian Journal of Pharmacology  2009;41(1):36-40.
Objectives:
To evaluate the hepatoprotective activity of ethanolic extract of Cleome viscosa Linn. (Capparidaceae) against carbon tetrachloride (CCI4) induced hepatotoxicity in experimental animal models.
Materials and Methods:
Leaf powder of Cleome viscosa was extracted with ethanol. The hepatoprotective activity of the extract was assessed in CCI4 induced hepatotoxicity in rats. Various biochemical parameters were estimated and histopathological studies were also performed on rat liver. The hepatoprotective activity was also supported by determining a functional parameter, i.e. thiopental-induced sleep of mice poisoned with CCl4.
Results:
The test material was found effective as hepatoprotective, through in vivo and histopathological studies. The extract was found to be effective in shortening the thiopental induced sleep in mice poisoned with CCl4. The hepatoprotective effect of ethanolic extract was comparable to that of silymarin, a standard hepatoprotective agent.
Conclusion:
The results of the present study show that ethanolic extract of Cleome viscosa has significant hepatoprotective activity.
doi:10.4103/0253-7613.48892
PMCID: PMC2825012  PMID: 20177580
Carbon tetrachloride; Cleome viscosa Linn; hepatoprotective
9.  Glycyrrhizic acid attenuates CCl4-induced hepatocyte apoptosis in rats via a p53-mediated pathway 
AIM: To investigate the effect of glycyrrhizic acid (GA) on carbon tetrachloride (CCl4)-induced hepatocyte apoptosis in rats via a p53-dependent mitochondrial pathway.
METHODS: Forty-five male Sprague-Dawley rats were randomly and equally divided into three groups, the control group, the CCl4 group, and the GA treatment group. To induce liver fibrosis in this model, rats were given a subcutaneous injection of a 40% solution of CCl4 in olive oil at a dose of 0.3 mL/100 g body weight biweekly for 8 wk, while controls received the same isovolumetric dose of olive oil by hypodermic injection, with an initial double-dose injection. In the GA group, rats were also treated with a 40% solution of CCl4 plus 0.2% GA solution in double distilled water by the intraperitoneal injection of 3 mL per rat three times a week from the first week following previously published methods, with modifications. Controls were given the same isovolumetric dose of double distilled water. Liver function parameters, such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen fibers were evaluated by Sirius red staining. Hepatocyte apoptosis was investigated using the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL) assay and the cleaved caspase-3 immunohistochemistry assay. The expression levels of p53 and apoptosis-related proteins were evaluated by immunohistochemistry or Western blotting analysis.
RESULTS: After 8 wk of treatment, GA significantly reduced serum activity of ALT (from 526.7 ± 57.2 to 342 ± 44.8, P < 0.05) and AST (from 640 ± 33.7 to 462.8 ± 30.6, P < 0.05), attenuated the changes in liver histopathology and reduced the staging score (from 3.53 ± 0.74 to 3.00 ± 0.76, P < 0.05) in CCl4-treated rats. GA markedly reduced the positive area of Sirius red and the ratio of the hepatic fibrotic region (from 7.87% ± 0.66% to 3.68% ± 0.32%, P < 0.05) compared with the CCl4 group. GA also decreased the expression level of cleaved caspase-3 compared to the CCl4 group. TUNEL assay indicated that GA significantly diminished the number of TUNEL-positive cells compared with the CCl4 group (P < 0.05). GA treatment clearly decreased the level of p53 (P < 0.05) detected by immunohistochemistry and Western blotting analysis. Compared with the CCl4 group, we also found that GA reduced the Bax/Bcl-2 ratio (P < 0.05), the expression of cleaved caspase-3 (P < 0.05), cleaved caspase-9 (P < 0.05), and inhibited cytochrome C and second mitochondria-derived activator of caspases (Smac) release from mitochondria to cytoplasm, i.e., GA reduced the expression level of Smac, which inhibited c-IAP1 activity (P < 0.05), ultimately inhibiting the activity of caspase-3, according to Western blotting analysis. As a result, GA suppressed activation of the caspase cascades and prevented hepatocyte apoptosis.
CONCLUSION: GA can inhibit CCl4-induced hepatocyte apoptosis via a p53-dependent mitochondrial pathway to retard the progress of liver fibrosis in rats.
doi:10.3748/wjg.v19.i24.3781
PMCID: PMC3699029  PMID: 23840116
P53; Apoptosis; Liver fibrosis; Glycyrrhizic acid; Mitochondria
10.  (Z)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione protects rats from carbon tetrachloride-induced liver injury and fibrogenesis 
AIM: To evaluate the hepatoprotective roles of (Z)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione (SKLB010) against carbon tetrachloride (CCl4)-induced acute and chronic liver injury and its underlying mechanisms of action.
METHODS: In the first experiment, rats were weighed and randomly divided into 5 groups (five rats in each group) to assess the protective effect of SKLB010 on acute liver injury. For induction of acute injury, rats were administered a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl4 dissolved in olive oil (1:1). Group 1 was untreated and served as the control group; group 2 received CCl4 for induction of liver injury and served as the model group. In groups 3, 4 and 5, rats receiving CCl4 were also treated with SKLB010 at doses of 25, 50 and 100 mg/kg, respectively. Blood samples were collected at 6, 12 and 24 h after CCl4 intoxication to determine the serum activity of alanine amino transferase. Tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β) were determined using enzyme-linked immunosorbent assay. At 24 h after CCl4 injection,liver fibrogenesis was evaluated by hematoxylin-eosin (HE) staining and immunohistochemical analyses. Cytokine transcript levels of TNF-α, IL-1β and inducible nitric oxide synthase in the liver tissues of rats were measured using a reverse transcriptase reverse transcription-polymerase chain reaction technique. In the second experiment, rats were randomly divided into 2 groups (15 rats in each group), and liver injury in the CCl4-administered groups was induced by a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl4 dissolved in olive oil (1:1). The SKLB010-treated groups received oral 100 mg/kg SKLB010 before CCl4 administration. Five rats in each group were sacrificed at 2 h, 6 h, 12 h after CCl4 intoxication and small fortions of livers were rapidly frozen for extraction of total RNA, hepatic proteins and glutathione (GSH) assays. In the hepatic fibrosis model group, rats were randomly divided into 2 groups (5 rats each group). Rats were injected intraperitoneally with a mixture of CCl4 (1 mL/kg body weight) and olive oil [1:1 (v/v)] twice a week for 4 wk. In the SKLB010-treated groups, SKLB010 (100 mg/kg) was given once daily by oral gavage for 4 wk after CCl4 administration. The rats were sacrificed one week after the last injection and the livers from each group were harvested and fixed in 10% formalin for HE and immunohistochemical staining.
RESULTS: In this rat acute liver injury model, oral administration of SKLB010 blocked liver tissue injury by down-regulating the serum levels of alanine aminotransferase, suppressing inflammatory infiltration to liver tissue, and improving the histological architecture of liver. SKLB010 inhibited the activation of NF-κB by suppressing the degradation of IκB, and prevented the secretion of pro-inflammatory mediators such as tumor necrosis factor-α, interleukin-1β, and the reactive free radical, nitric oxide, at the transcriptional and translational levels. In this chronic liver fibrosis model, treatment with 100 mg/kg per day SKLB010 attenuated the degree of hepatic fibrosis and area of collagen, and blocked the accumulation of smooth-muscle actin-expressed cells.
CONCLUSION: These results suggest that SKLB010 is a potent therapeutic agent for the treatment of CCl4-induced hepatic injury.
doi:10.3748/wjg.v18.i7.654
PMCID: PMC3281222  PMID: 22363136
Anti-inflammatory effects; Anti-oxidative effects; (Z)-5-(4-methoxybenzylidene) thiazolidine-2; 4-dione (SKLB010) against carbon tetrachloride; Fibrogenesis; Hepatitis; Nuclear factor-κB; SKLB010
11.  Antioxidant and Protective Effect of Ethyl Acetate Extract of Podophyllum Hexandrum Rhizome on Carbon Tetrachloride Induced Rat Liver Injury 
The antioxidant and hepatoprotective activities of ethyl acetate extract was carefully investigated by the methods of DPPH radical scavenging activity, Hydroxyl radical scavenging activity, Superoxide radical scavenging activity, Hydrogen peroxide radical scavenging activity and its Reducing power ability. All these in vitro antioxidant activities were concentration dependent which were compared with standard antioxidants such as BHT, α-tocopherol. The hepatoprotective potential of Podophyllum hexandrum extract was also evaluated in male Wistar rats against carbon tetrachloride (CCl4)-induced liver damage. Pre-treated rats were given ethyl acetate extract at 20, 30 and 50 mg/kg dose prior to CCl4 administration (1 ml/kg, 1:1 in olive oil). Rats pre-treated with Podophyllum hexandrum extract remarkably prevented the elevation of serum AST, ALT, LDH and liver lipid peroxides in CCl4-treated rats. Hepatic glutathione levels were significantly increased by the treatment with the extract in all the experimental groups. The extract at the tested doses also restored the levels of liver homogenate enzymes (glutathione peroxidase, glutathione reductase, superoxide dismutase and glutathione-S- transferase) significantly. This study suggests that ethyl acetate extract of P. hexandrum has a liver protective effect against CCl4-induced hepatotoxicity and possess in vitro antioxidant activities.
doi:10.1155/2011/238020
PMCID: PMC3025527  PMID: 21394192
12.  Hepatoprotective activity of petroleum ether, diethyl ether, and methanol extract of Scoparia dulcis L. against CCl4-induced acute liver injury in mice 
Indian Journal of Pharmacology  2009;41(3):110-114.
Objectives:
The present study was aimed at assessing the hepatoprotective activity of 1:1:1 petroleum ether, diethyl ether, and methanol (PDM) extract of Scoparia dulcis L. against carbon tetrachloride-induced acute liver injury in mice.
Materials and Methods:
The PDM extract (50, 200, and 800 mg/kg, p.o.) and standard, silymarin (100 mg/kg, p.o) were tested for their antihepatotoxic activity against CCl4-induced acute liver injury in mice. The hepatoprotective activity was evaluated by measuring aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and total proteins in serum, glycogen, lipid peroxides, superoxide dismutase, and glutathione reductase levels in liver homogenate and by histopathological analysis of the liver tissue. In addition, the extract was also evaluated for its in vitro antioxidant activity using 1, 1-Diphenyl-2-picrylhydrazyl scavenging assay.
Results:
The extract at the dose of 800 mg/kg, p.o., significantly prevented CCl4-induced changes in the serum and liver biochemistry (P < 0.05) and changes in liver histopathology. The above results are comparable to standard, silymarin (100 mg/kg, p.o.). In the in vitro 1, 1-diphenyl-2-picrylhydrazyl scavenging assay, the extract showed good free radical scavenging potential (IC 50 38.9 ± 1.0 μg/ml).
Conclusions:
The results of the study indicate that the PDM extract of Scoparia dulcis L. possesses potential hepatoprotective activity, which may be attributed to its free radical scavenging potential, due to the terpenoid constituents.
doi:10.4103/0253-7613.55206
PMCID: PMC2861810  PMID: 20442817
Carbon tetrachloride; hepatoprotective activity; Scoparia dulcis L.
13.  Hepatoprotective effects of Rubus coreanus miquel concentrates on liver injuries induced by carbon tetrachloride in rats 
As well-being foods pursuing healthy life are becoming popular, interest in Rubus coreanus Miquel (RCM) fruit, a type of Korean blackberry, is increasing due to its medicinal actions including protecting the liver, brightening the eyes, and alleviating diabetes. This study was carried out to evaluate the hepatoprotective effects of RCM concentrates on liver injuries induced by carbon tetrachloride (CCl4) in rats. RCM, produced in June ~ July 2008 at Chunbook, Gochang (South Korea), was finely mashed. The seeds were removed and the juices were condensed. Thirty-two Sprague-Dawley rats were divided into four groups according to treatment: normal (eight rats), CCl4, 1% RCM, and 2% RCM. Experimental diets were provided to the experimental animals for 4 weeks. We measure total cholesterol, high density lipoprotein-cholesterol (HDL-C), aspartate amino transferase (AST), alanine amino transferase (ALT), and alkaline phosphatase (ALP) levels. Part of the livers was isolated for histopathological evaluation, and analyzed for lipid peroxide (TBARS), superoxide dismutase (SOD) and liver proteins. The activities of serum AST, ALT, and ALP were elevated following CCl4 administration. Levels of hepatic TBARS were also significantly increased in the CCl4 groups. However, hepatic TBARS levels and the activities of serum enzymes were markedly reduced by supplementation with the RCM concentrates (P < 0.05). Hepatic SOD activity increased in the RCM concentrates group versus CCl4 groups. Histopathological examination revealed massive necrosis in the centrilobular area and degenerative changes caused by CCl4 were ameliorated by dietary supplementation with RCM concentrates. These results suggest that RCM concentrates have hepatoprotective effects and may improve the symptoms of liver injuries.
doi:10.4162/nrp.2014.8.1.40
PMCID: PMC3944155  PMID: 24611104
Rubus coreanus Miquel; carbon tetrachloride; hepatoprotective effects
14.  Hepatoprotective activity of Symplocos racemosa bark on carbon tetrachloride-induced hepatic damage in rats 
The present study aims to evaluate the hepatoprotective activity of ethanol extract of Symplocos racemosa (EESR) bark on carbon tetrachloride (CCl4)-induced hepatic damage in rats. CCl4 with olive oil (1 : 1) (0.2 ml/kg, i.p.) was administered for ten days to induce hepatotoxicity. EESR (200 and 400 mg/kg, p.o.) and silymarin (100 mg/kg p.o.) were administered concomitantly for fourteen days. The degree of hepatoprotection was measured using serum transaminases (AST and ALT), alkaline phosphatase, bilirubin, albumin, and total protein levels. Metabolic function of the liver was evaluated by thiopentone-induced sleeping time. Antioxidant activity was assessed by measuring liver malondialdehyde, glutathione, catalase, and superoxide dismutase levels. Histopathological changes of liver sample were also observed. Significant hepatotoxicity was induced by CCl4 in experimental animals. EESR treatment showed significant dose-dependent restoration of serum enzymes, bilirubin, albumin, total proteins, and antioxidant levels. Improvements in hepatoprotection and morphological and histopathological changes were also observed in the EESR treated rats. It was therefore concluded that EESR bark is an effective hepatoprotective agent in CCl4-induced hepatic damage, and has potential clinical applications for treatment of liver diseases.
doi:10.4103/0975-9476.85552
PMCID: PMC3193685  PMID: 22022156
Carbon tetrachloride; hepatoprotective; Symplocos racemosa; silymarin
15.  Hepatoprotective activity of Leptadenia reticulata stems against carbon tetrachloride-induced hepatotoxicity in rats 
Indian Journal of Pharmacology  2011;43(3):254-257.
Objective:
To evaluate the hepatoprotective activity of ethanolic and aqueous extract of stems of Leptadenia reticulata (Retz.) Wight. and Arn. in carbon tetrachloride (CCl4)-induced hepatotoxicity in rats.
Materials and Methods:
The toxicant CCl4 was used to induce hepatotoxicity at a dose of 1.25 ml/kg as 1 : 1 mixture with olive oil. Ethanolic and aqueous extracts of L. reticulata stems were administered in the doses of 250 and 500 mg/kg/day orally for 7 days. Silymarin (50 mg/kg) was used as standard drug. The hepatoprotective effect of these extracts was evaluated by the assessment of biochemical parameters such as serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, alkaline phosphatase, total bilirubin, serum protein, and histopathological studies of the liver.
Results:
Treatment of animals with ethanolic and aqueous extracts significantly reduced the liver damage and the symptoms of liver injury by restoration of architecture of liver as indicated by lower levels of serum bilirubin and protein as compared with the normal and silymarin-treated groups. Histology of the liver sections confirmed that the extracts prevented hepatic damage induced by CCl4 showing the presence of normal hepatic cords, absence of necrosis, and fatty infiltration.
Conclusion:
The ethanolic and aqueous extracts of stems of L. reticulata showed significant hepatoprotective activity. The ethanolic extract is more potent in hepatoprotection in CCl4-indiced liver injury model as compared with aqueous extract.
doi:10.4103/0253-7613.81507
PMCID: PMC3113374  PMID: 21713086
Carbon tetrachloride; hepatoprotective activity; Leptadenia reticulata; silymarin
16.  Evaluation of Hepatoprotective Effect of Leaves of Cassia sophera Linn. 
In the present study, the hepatoprotective activity of ethanolic extracts of Cassia sophera Linn. leaves was evaluated against carbon-tetrachloride- (CCl4-) induced hepatic damage in rats. The extracts at doses of 200 and 400 mg/kg were administered orally once daily. The hepatoprotection was assessed in terms of reduction in histological damage, changes in serum enzymes, serum glutamate oxaloacetate transaminase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP), total bilirubin, and total protein levels. The substantially elevated serum enzymatic levels of AST, ALT, ALP, and total bilirubin were restored towards the normalization significantly by the extracts. The decreased serum total protein level was significantly normalized. Silymarin was used as standard reference and exhibited significant hepatoprotective activity against carbon tetrachloride-induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that Cassia sophera leaves have potent hepatoprotective action against carbon tetrachloride-induced hepatic damage in rats. This study suggests that possible activity may be due to the presence of flavonoids in the extracts.
doi:10.1155/2012/436139
PMCID: PMC3368335  PMID: 22690244
17.  Graptopetalum Paraguayense Ameliorates Chemical-Induced Rat Hepatic Fibrosis In Vivo and Inactivates Stellate Cells and Kupffer Cells In Vitro 
PLoS ONE  2013;8(1):e53988.
Background
Graptopetalum paraguayense (GP) is a folk herbal medicine with hepatoprotective effects that is used in Taiwan. The aim of this study was to evaluate the hepatoprotective and antifibrotic effects of GP on experimental hepatic fibrosis in both dimethylnitrosamine (DMN)- and carbon tetrachloride (CCl4)-induced liver injury rats.
Methods
Hepatic fibrosis-induced rats were fed with the methanolic extract of GP (MGP) by oral administration every day. Immunohistochemistry, biochemical assays, and Western blot analysis were performed. The effects of MGP on the expression of fibrotic markers and cytokines in the primary cultured hepatic stellate cells (HSCs) and Kupffer cells, respectively, were evaluated.
Results
Oral administration of MGP significantly alleviated DMN- or CCl4-induced liver inflammation and fibrosis. High levels of alanine transaminase, aspartate transaminase, bilirubin, prothrombin activity and mortality rates also decreased in rats treated with MGP. There were significantly decreased hydroxyproline levels in therapeutic rats compared with those of the liver-damaged rats. Collagen I and alpha smooth muscle actin (α-SMA) expression were all reduced by incubation with MGP in primary cultured rat HSCs. Furthermore, MGP induced apoptotic cell death in activated HSCs. MGP also suppressed lipopolysaccharide-stimulated rat Kupffer cell activation by decreasing nitric oxide, tumor necrosis factor-α and interleukin-6 production, and increasing interleukin-10 expression.
Conclusions
The results show that the administration of MGP attenuated toxin-induced hepatic damage and fibrosis in vivo and inhibited HSC and Kupffer cell activation in vitro, suggesting that MGP might be a promising complementary or alternative therapeutic agent for liver inflammation and fibrosis.
doi:10.1371/journal.pone.0053988
PMCID: PMC3545924  PMID: 23335984
18.  Hepatoprotective effects of berberine on carbon tetrachloride-induced acute hepatotoxicity in rats 
Chinese Medicine  2010;5:33.
Background
Berberine is an active compound in Coptidis Rhizoma (Huanglian) with multiple pharmacological activities including antimicrobial, antiviral, anti-inflammatory, cholesterol-lowering and anticancer effects. The present study aims to determine the hepatoprotective effects of berberine on serum and tissue superoxide dismutase (SOD) levels, the histology in tetrachloride (CCl4)-induced liver injury.
Methods
Sprague-Dawley rats aged seven weeks were injected intraperitoneally with 50% CCl4 in olive oil. Berberine was orally administered before or after CCl4 treatment in various groups. Twenty-four hours after CCl4 injection, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, serum and liver superoxide dismutase (SOD) activities were measured. Histological changes of liver were examined with microscopy.
Results
Serum ALT and AST activities significantly decreased in a dose-dependent manner in both pre-treatment and post-treatment groups with berberine. Berberine increased the SOD activity in liver. Histological examination showed lowered liver damage in berberine-treated groups.
Conclusion
The present study demonstrates that berberine possesses hepatoprotective effects against CCl4-induced hepatotoxicity and that the effects are both preventive and curative. Berberine should have potential for developing a new drug to treat liver toxicity.
doi:10.1186/1749-8546-5-33
PMCID: PMC2949732  PMID: 20849653
19.  Protective effects of Launaea procumbens on rat testis damage by CCl4 
Background
Traditionally various human diseases of kidneys, hormonal imbalance and sexual diseases are treated with Launaea procumbens (L). In the present study protective effects of methanolic extract of Launaea procumbens (LPME) was evaluated against CCl4-induced oxidative damages in rat testis.
Methods
To examine the protective effects of Launaea procumbens on testis against oxidative stress of carbon tetrachloride in male rat, 30 male albino rats were equally divided into 5 groups (6 rats). First group was given standard diet and drinking water. Second group received CCl4 3 ml/kg intraperitoneally (30% in olive oil). Third and forth were given orally 100; 200 mg/kg b.w., in 99.8% dimethyl sulphooxide (DMSO), Launaea procumbens methanolic extracts (LPME) after 48 h of CCl4 treatment twice a week and sixth group received only LPME in DMSO at a dose of 200 mg/kg b.w., for four weeks. Protective effects of Launaea procumbens were observed on sperm concentration, motility and morphology, serum reproductive hormonal level, activity of antioxidant enzymes, lipid peroxidation (TBARS) and DNA damages.
Results
Results of the present study revealed that treatment of CCl4 significantly (p < 0.01) reduced sperm concentration and motility comparatively to controls. Level of testosterone, luteinizing hormone and follicle stimulating hormone, were depleted markedly (p <0.01) with treatment of CCl4. In addition, CCl4 induction in rats reduced activities of antioxidant enzymes while increased lipid peroxidation and DNA damages. Co-administration of LPME significantly (p <0.01) improved these alterations in improving of hormonal level, activities of antioxidant enzymes and lipid peroxidation near to control rats.
Conclusion
From the results it is suggested that Launaea procumbens methanolic extract has the ability to protect testis against oxidative damages, possibly through antioxidant effects of its bioactive compounds.
doi:10.1186/1476-511X-11-103
PMCID: PMC3494576  PMID: 22913836
Launaea procumbens; GSH; CCl4; Lipidperoxidation; DNA damages
20.  Hepatoprotective activity of Hepax-A polyherbal formulation 
Objective
To evaluate the hepatoprotective potential of Hepax, a polyherbal formulation, against three experimentally induced hepatotoxicity models in rats.
Methods
Hepatoprotective activity of Hepax was studied against three experimentally induced hepatotoxicity models, namely, carbon tetrachloride (CCl4), paracetamol and thiocetamide induced hepatotoxicity in rats.
Results
Administration of hepatotoxins (CCl4, paracetamol and thiocetamide) showed significant morphological, biochemical and histological deteriorations in the liver of experimental animals. Pretreatment with Hepax had significant protection against hepatic damage by maintaining the morphological parameters (liver weight and liver weight to organ weight ratio) within normal range and normalizing the elevated levels of biochemical parameters (SGPT, SGOT, ALP and total bilirubin), which were evidently showed in histopathological study.
Conclusions
The Hepax has highly significant hepatoprotective effect at 100 and 200 mg/kg, p.o. on the liver of all the three experimental animal models.
doi:10.1016/S2221-1691(11)60013-0
PMCID: PMC3609179  PMID: 23569745
Hepax; Paracetamol; CCl4; Thiocetamide; Hepatotoxicity
21.  Protective Activity of the Stem Bark Aqueous Extract of Musanga Cecropioides in Carbon Tetrachloride- and Acetaminophen-induced Acute Hepatotoxicity in Rats 
The hepatoprotective activities and the mechanisms of actions of Musanga cecropioides stem bark aqueous extract (MCW) were investigated on acute hepatocellular injuries induced by intraperitoneal (IP) carbon tetrachloride (CCl4) (20% CCl4/olive oil, 1.5 mL/kg) and 800 mg/kg/IP of acetaminophen (APAP) in normal saline, in male Wistar rats. Among the Yorubas (South-West Nigeria), cold decoction of MCW is used as a natural antidote for oral gastric poisonings, infective hepatitis and other liver diseases. Its hepatoprotective activities were monitored by assaying for the serum aminotransferases, ornithine carbamoyl transferase and the toxicant-induced histopathological lesions in rat livers 24 hours post-induction. These enzymes are markers of acute hepatic injuries and their elevations are indications of acute liver injuries. Pretreatment of rats with graded doses (125 – 500 mg/kg) of MCW significantly attenuated the acute elevation of the liver enzymes and the hepatotoxin-induced histopathological lesions in the rat livers. The presence of two active natural antioxidants (flavonoids and alkaloids) in high concentrations in MCW may account for the hepatoprotective activities observed in this study. These results, thus, support the folkloric use of MCW for treatment of hepatic injuries resulting from acute gastric poisonings, infective hepatitis or other liver diseases.
PMCID: PMC2816566  PMID: 20209004
Musanga cecropioides; Hepatoprotective activities; Carbon tetrachloride; Acetaminophen; Wistar rats
22.  Hepatoprotective Role of Ethanolic Extract of Vitex negundo in Thioacetamide-Induced Liver Fibrosis in Male Rats 
The hepatoprotective activity of ethanolic extract from the leaves of Vitex negundo (VN) was conducted against thioacetamide- (TAA-) induced hepatic injury in Sprague Dawley rats. The therapeutic effect of the extract was investigated on adult male rats. Rats were divided into seven groups: control, TAA, Silymarin (SY), and VN high dose and low dose groups. Rats were administered with VN extract at two different doses, 100 mg/kg and 300 mg/kg body weight. After 12 weeks, the rats administered with VN showed a significantly lower liver to body weight ratio. Their abnormal levels of biochemical parameters and liver malondialdehyde were restored closer to the normal levels and were comparable to the levels in animals treated with the standard drug, SY. Gross necropsy and histopathological examination further confirmed the results. Progression of liver fibrosis induced by TAA in rats was intervened by VN extract administration, and these effects were similar to those administered with SY. This is the first report on hepatoprotective effect of VN against TAA-induced liver fibrosis.
doi:10.1155/2013/739850
PMCID: PMC3671533  PMID: 23762157
23.  Protective Effect of Ssanghwa-Tang Fermented by Lactobacillus Fermentum Against Carbon Tetrachloride-Induced Acute Hepatotoxicity in Rats 
Ssanghwa-tang (SHT) is a traditional herbal medicine formula that has been used for the development of physical strength, relief of pain, and the reduction of fatigue. In this study, we fermented SHT with Lactobacillus fermentum (L. fermentum), Lactobacillus gasseri (L.gasseri), or Lactobacillus casei (L.casei) to investigate the hepatoprotective effects of SHT and fermented SHT with Lactobacillus on carbon tetrachloride (CCl4)-induced liver injury in rats. Rats were given CCl4 (1 ml/kg, 50% CCl4 in olive oil) intraperitoneally and either SHT or fermented SHTs (15 ml/kg) was administered 30 min before CCl4. At 24 hr after CCl4 injection, the levels of transaminases in the serum were markedly increased. These increases were significantly attenuated by either SHT + L. fermentum or SHT+ L.gasseri. However, SHT and SHT + L.casei showed slight suppression of the increase of transaminases. The liver histological changes were diminished by treatment with SHT + L. fermentum. Additionally, the potential hepatoprotective effect of fermented-SHTs correlated with the amount of unknown metabolite which is produced during fermentation process with L. fermentum, L.gasseri, or L.casei. Therefore, these results suggest that the hepatoprotective effect of SHT may be improved by fermentation with L. fermentum and the intestinal bacterial enzyme activities may likely play an important role in the pharmacological action of herbal medicines.
PMCID: PMC3252232  PMID: 22468011
Carbon tetrachloride; Fermentation; Herbal fomula; Lactobacillus; Ssanghwa-tang
24.  Hepatoprotective Activity of the Ethanolic Extract of Ficus caricaLinn. LeavesinCarbon Tetrachloride-Induced Hepatotoxicityin Rats 
The ethanolic extract of Ficus carica leaves was screened for hepatoprotective and antioxidant activity in hepatotoxic Albino rats induced via carbon tetrachloride. The degree of protection was measured by estimating biochemical parameters such as serum glutamate Oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase ( SGPT),totalprotein (TP), totalalbumin (TA), alkaline phosphatase (ALKP) and the level of total serum bilirubin. The extract in addition reduced CCl4 induced lipid peroxidation in-vivo and in-vitro. The ethanolic extract (50 mg/kg, 100mg/kg,200mg/kg)exhibited significant hepatoprotection incarbontetra chloride in toxicated rats in a dose dependant manner. The hepatoprotective effects of the extract were comparable with the standard drug silymarin 10)mg/kg body weight, IP).
PMCID: PMC3828912  PMID: 24250358
Ficus carica; Hepatoprotective activity; Carbontetra chloride; Silymarin
25.  Hepatoprotective effect of poly herbal formulation against various hepatotoxic agents in rats 
Pharmacognosy Research  2012;4(1):50-56.
Background:
Individually Andrographis paniculata Nees. (Acanthaceae), Phyllanthus niruri Linn.(Euphorbiaceae) and Phyllanthus emblica Linn. single plant extracts have been reported to have hepatoprotective activity. However, literature survey shows that no sufficient scientific data has been publish on pharmacological evaluation of these plants in combined form.
Method:
Hepatoprotective activity of the polyherbal hepatoprotaective formulation (PHF)-containing spray-dried aqueous extracts of Andrographis paniculata Nees. (Acanthaceae), Phyllanthus niruri Linn. (Euphorbiaceae) and Phyllanthus emblica Linn. (Euphorbiaceae), was screened against paracetamol, carbon tetrachloride (CCl4), and ethanol-induced hepatic damage in rats. PHF was evaluated by measuring levels of serum marker enzymes like SGOT, SGPT, ALP, direct bilirubin (DB), and lactate dehydrogenase (LDH). The histological studies were also studied support the biochemical parameters. Silymarin was used as standard drug.
Results:
Administration of PHF (100 and 200 mg/kg p.o.) significantly inhibited paracetamol, CCl4 and ethanol-induced elevation levels of SGPT, SGOT, ALP, DB and LDH. A comparative histopathological study of liver exhibited almost normal architecture as compared to toxicant group.
Conclusion:
Results suggests that the hepatoprotective effects of PHF might be useful for liver protection due to combined action of all plant extracts along with their phytoconstituents.
doi:10.4103/0974-8490.91040
PMCID: PMC3250040  PMID: 22224062
Andrographis paniculata Nees; carbon tetrachloride; hepatoprotective activity; Marker enzymes; paracetamol; Phyllanthus niruri Linn

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