CCl4-induced Hepatotoxicity Research Articles

Ablation of apoptosis-stimulating of p53 protein 1 protects mice from acute hepatic injury and dysfunction via NF-κB pathway in CCl4-induced hepatotoxicity

Abstract Acute liver injury (ALI) is characterized by apoptosis, inflammation, and oxidative stress, and pathogenic mechanism of ALI is poorly understood. Apoptosis-stimulating of p53 protein 1 (ASPP1) is involved in environmental responses, tumor growth, and NF-KB activity, which is of critical importance to ALI. However, the role of ASPP1 in ALI remains largely unexplored. The current study aimed to determine the role of ASPP1 in ALI induced by CCl4 and the underlying mechanism. ASPP1 expression was detected in wild type (WT) mice with ALI induced by CCl4. The function of ASPP1 in ALI induced by CCl4 was investigated using conventional knockout ASPP1 mice. ASPP1 expression significantly increased in ALI mice at 24 hours after CCl4 injection. Deletion of ASSP1 ameliorated apoptosis, inflammation, and necrosis in ALI relative to WT mice. In addition, deficiency of ASPP1 improved liver flood flow as well as ALT and AST levels. The levels of phosphorylated p65 and phosphorylated IκBα were lower in ASPP1-/- mice than in WT mice with ALI. These results implicate that deletion of ASPP1 may act via inhibition of the NF-κB pathway and protect mice from ALI, which may be a new potential therapeutic target for the treatment of ALI.

EFFECT OF CHICORY (CICHORIUM INTYBUS L.) LEAVES EXTRACT TO PROTECT CERTAIN LIVER ENZYMES IN MICE AGAINST CARBON TETRACHLORIDE-INDUCED HEPATOTOXICITY

Chicory contains dietary inulin and oligofructose compounds which have a beneficial effect on metabolism, as well as polyphenol type derivatives, which may be responsible for the antioxidant properties. The effect of different concentrations of the hydroalcoholic extract of dried powdered leaves of chicory on CCl4-induced hepatotoxicity in vivo in mice, has been studied. Thirty-five mice (20-30 g) of age four weeks were randomly divided into 5 groups (seven in each). The mice orally received three different concentrations of chicory leaves extract (CLE) before injection for 3 consecutive days by CCl4. The study showed that CCl4 caused elevation in serum GOT, GPT, and ALP activities in mice, but there was no pronounced elevation in total protein and albumin. Pretreatment of mice with 75, 150, and 300 mg/kg/bwt of CLE protected the liver against CCl4-induced cytotoxicity. CLE significantly (p<0.05) reduced the elevated level of cholesterol, triglycerides, and significantly increased the levels of HDL in the group of mice pretreated with CLE (150 mg/ kg); 145± 1.3, 243± 1.3, and 34± 1.2 mg/dl, respectively, compared with negative control (185± 1.4, 290± 1.4, and 21± 1.3 mg/dl, respectively). In contrast, serum fasting glucose and magnesium level in mice groups were not significantly different under the same conditions. CLE is potentially a good natural source for antioxidants and natural hepatoprotective as well as beneficial to who is suffering from hypercholesterolemia.

Therapeutic potential of Ni(II) Schiff base complex on CCl4 toxicity

Carbon tetrachloride (CCl4) is a known ecological hazardous xenobiotic that could motivate hepatotoxicity. We aimed to examine, for the first time, the therapeutic potential of nickel(II) diacetyl monoxime-2-pyridyl hydrazone complex versus CCl4-induced hepatotoxicity in rats. We used six rat groups of ten animals each. The negative control, vehicle, normal rats injected i.p. with the complex (2.4 mg/kg/day), positive control rats injected i.p. with CCl4, and treated rats administered complex via injection at low and high concentrations of 1.2 and 2.4 mg/kg/day, respectively at the same time as CCl4 injection for short-term (3-weeks) and long-term (8-weeks) treatment. Intoxicated-rodents exhibited significant elevations in the liver index, hepatic serum markers, and oxidative stress with significant reductions in the hepatic, antioxidants, nucleic acids, and proteins. Complex co-treatment with CCl4 significantly suppressed the elevated liver enzyme activities, attenuated oxidative stress, reactivated antioxidant-system components, and amended the hepatic tissue injury. The complex high dose was more efficient than the low dose. Results of negative control were analogous to those of normal rats injected with the complex high dose. The histopathological analysis also supported the above findings. These results show that complex has good antioxidant and therapeutic properties, which can help in treating and preventing CCl4-induced hepatotoxicity.

Preclinical Study on the Hepatoprotective Effect of Pollen Extract of Pinus brutia Ten. (Red Pine) in Mice and Phenolic Acid Analysis

Many agents, including those from herbal sources, have been sought as preventives or cures for hepatotoxicity. The pollen of Pinus brutia Ten., known as red pine (Pinaceae), is used against liver diseases in Anatolian folk medicine. In the current study, pollen ethanol extract of P. brutia was investigated for its possible hepatoprotective activity using a mouse model of CCl4-induced hepatotoxicity. Swiss albino mice were divided into five groups, and extract-treated groups were compared with a silymarin-treated group as the reference. The extract was tested at 100, 200, and 300 mg/kg (b.w.). Phenolic acids were analyzed using high-performance column chromatography (HPLC) in the extracts as pollens are usually known to be rich in phenolics. Our data revealed that the extract displayed the best hepatoprotection at a dose of 100 mg/kg when compared with silymarin (Legalon®), the reference drug. HPLC analysis indicated presence of protocatechuic acid (0.176 mg/g extract), p-hydroxybenzoic acid (0.001 mg/g extract), vanillic acid (VA) (0.537 mg/g extract), syringic acid (0.050 mg/g extract), and tr-cinnamic acid (0.310 mg/g extract), while the major phenolic acid was VA. The outcomes of this study allow us to conclude that red pine pollen extract can serve as a promising hepatoprotective agent. Among the phenolic acids analyzed in the pollen extract, vanillic acid as the major one besides some other phenolic acids detected seems to be responsible for its remarkable hepatoprotective effect.

English

The aim of this study was to evaluate the effect of Berberis vulgaris water extract (BWE) against CCl4-induced hepatotoxicity and LPS/PCM induced- hepatic inflammation in the experimental animal models. CCl4 (100 ug/kg, oral) administration for 28 days as well as the co-administration of LPS (250 ug/kg bw) and PCM (2 g/kg bw) for 28 days resulted in massive elevation in serum and hepatic prooxidants and inflammatory parameters (thiobarbituric acid reactive substances (TBARS), tissue tumor necrosis factor–alpha (TNF-α), interleukin 6 (IL-6) and IL-12 and nitric oxide (NO) with depletion in antioxidants system (superoxide dismutase (SOD) and glutathione peroxidase (GPx). These alterations were combined with elevation of serum liver function tests (alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase level and (ALP). Oral administration of the BWE (100 mg/kg) for 15 days showed hepatoprotective and anti-inflammatory effects against both CCl4 and LPS/PCM as it increased the activities of antioxidants enzymes, decreased the prooxidants and inflammatory markers levels and improved the serum liver function tests levels. The obtained findings thus suggest that B. vulgaris aqueous extract has protective roles against hepatic toxicity and hepatic inflammation due to its antioxidant and anti-inflammatory properties. Key words: Carbon tetrachloride, paracetamol, lipopolysaccharide, glutathione peroxidase, alanine transferase.

Hepatoprotective activity of Phyllanthus niruri Linn. endophytes

BackgroundThe liver is the vital organ which plays a major role in metabolism with numerous functions in the human beings such as protein synthesis, hormone production, and detoxification. Present research work is focused on hepatoprotective potential of chloroform (PNFC) and ethyl acetate (PNFEA) endophytic fractions from Phyllanthus niruri Linn. against CCl4-induced hepatotoxicity in albino Wistar rats. To test our hypothesis, both endophytic fungal fractions were tested for vitro antioxidant and in vivo hepatoprotective activity. Serum biochemical parameters like SGOT, SGOT, SALP, cholesterol, bilirubin, and protein were estimated to assess hepatoprotective activity.ResultsGroup of rats treated with CCl4 possess marked hepatic damage and oxidative stress which indicates that cellular leakage and loss of functional integrity of cell membrane in liver. PNFC and PNFEA fractions of endophyte from Phyllanthus niruri Linn. stem have significantly reduced the elevated levels of biomarkers like SGPT, SGOT, SALP, bilirubin, cholesterol, and total protein in CCl4-induced hepatotoxicity in rats. The results obtained confirm hepatoprotective activity of endophytic fractions (PNFC and PNFEA) mediated through the stabilization of plasma membrane, repair of hepatic tissue damage, return of biochemical marker levels to normal, and regeneration of hepatocytes. Histopathological observations revealed improvement in the liver architecture after the treatment of secondary metabolites of endophytic fractions against CCl4-induced liver damage. Both fungal endophytes PNFC and PNFEA showed DPPH scavenging activity with IC50 of 97.79 μg/ml and 108.40 μg/ml, respectively, and possess antioxidant potential. Presence of flavonoids in the both fractions of endophytes may be a possible reason for its antioxidant potential and identified as Eurotium amstelodami strain.ConclusionBoth fungal endophytes PNFC and PNFEA possess hepatoprotective potential due to the presence of secondary metabolites of fungi, i.e., Eurotiumam stelodami strain which support the claim endophytes and act as a potent biomedicine for treatment of various chronic diseases.

Bioactive Compounds from Mango peels (Mangifera indica .Tommy Atkins) And Demonstration of its Cytotoxicity and CCl4 Induced Hepatotoxicity in Rat

Mangifera indica (Tommy Atkins) peels, commonly known as mango, is a pharmacologically, ethnomedically, and phytochemically diverse plant. Peels is a major by-product during processing of mango fruit into pulp. In the present study, Thirteen pure bioactive compounds were isolated from methanolic peels extract. Six of them are new ellagitannins,namely,1,2,3,4,6 -Penta-O-galloyl-s-4C1-glupyranose (3); 2,3,6-Tri-O-galloyl-(α/β)-4C1-glucopyranose(4); 2,3-Di-O-galloyl-(α/β)-4C1-glucopyranose,Nilocitin(6);3,6-Di-O-galloyl-(α/β)-4C1-glucopyranose(8);1,6 -Di-O-galloyl- β-4C1-glucopyranose (9) ; 1,3-Di-O-galloyl-β-4C1- glucose (11), which were analyzed for the first time from M. indica (Tommy Atkins) peels. The ameliorative effect of the methanolic extract of M. indica (Tommy Atkins) peels towards the CCl4-induced hepatotoxicity in male Wistar rats through measuring certain biochemical parameters content in the liver were analyzed. The CCl4-treated rats showed a significant decline in the studied the serum levels of high-density lipoprotein (HDL), albumin (A) as well as the hepatic levels of glutathione (GSH) and activities of catalase (CAT), superoxide dismutase (SOD) , glutathione reductase (GR), elevation in the levels of total lipids (TL), triglycerides (TG), total cholesterol (TC), low-density lipoproteins (LDL), globulin (G), total bilirubin (TBil) , alanine and aspartate aminotransferase and alkaline phosphatase (ALAT and ASAT, ALP) and the hepatic levels of malondialdehyde (MDA). In contrast, the administration of methanol extract, notably improved all the studied parameters. This study showed that CCl4 administration to Wistar rats, at a high dose level, could induce a hepatic injury in addition to certain metabolic alterations. The work was extended to investigate tissue histopathology. Thus, results suggest that the peels extract can be a potential source of an attractive candidate for ameliorating of hepatotoxicity induced by CCl4 through scavenging free radicals, improved liver functions, and normalizing the liver histopathological architecture.

Effects of Aqueous Stem Bark Extract of Stereospermum kunthianum on Carbon Tetrachloride - Induced Hepatotoxicity in Rats

Aim: This study is aimed at evaluating the effects of the aqueous stem bark extract of Stereospermum kunthianum on CCl4-induced hepatotoxicity in rats.
 Methodology: Analysis of qualitative phytochemical components and antioxidant activity were carried out. Experimental rats were randomly divided into six groups of five rats each. Group 1: served as the normal control group. Group 2: was administered with CCl4 only at a dose of 3 ml/kg b.wt by single intraperitoneal administration. Group 3: served as the standard control group. Group 4: was administered with 200 mg/kg b.wt of the aqueous stem bark extract + CCl4. Group 5: was administered with 400 mg/kg b.wt of the aqueous stem bark extract + CCl4. Group 6: was administered with 600 mg/kg b.wt of the aqueous stem bark extract+ CCl4.
 Results: The phytochemical analysis showed the presence of flavonoid, phenols, saponins, and terpenoid while tannins and alkaloids were absent. The antioxidant activity showed that the extract significantly (P<0.05) inhibits Ferric Reducing Antioxidant Power (FRAP) and Thiobarbituric Acid Reactive Substances (TBARS), giving high activity as the concentration of the extract increases. The elevated levels of ALT and AST coupled with Conjugated bilirubin, Total bilirubin, and total protein caused by CCl4 administration were all reduced significantly (P<0.05) by the extract in dose dependent manner.
 Conclusion: These findings demonstrated that stem bark extract of Stereospernmum kunthianum could be an alternative medication for liver injury.

Heliconia rostrata rhizomes mitigate chemical-induced liver injury by debilitating oxidative stress in HepG2 cells and rats

Ethnopharmacological relevanceHeliconia rostrata Ruiz and Pav. belongs to the family Heliconiaceae. Plant was traditionally used to cure jaundice, intestinal pain, diabetes and hypertension. Aim of the experimentPresent study evaluated hepatoprotective efficacy of ethanol (REE) and methanol (RME) extracts of H. rostrata rhizomes in HepG2 cell lines and rats. Antioxidant efficacy of extracts was determined using ex vivo and in vivo methods. Materials and methodsBefore conducting efficacy studies, safety of REE and RME was established using toxicity studies which included Oral acute-fixed dose toxicity using OECD TG420, 28-days repeated dose oral toxicity by OECD TG407 and cytotoxicity studies by brine shrimp lethality (BSL) bioassay and MTT assay taking HepG2 cell line. Ex vivo (Extracts: 0–250 μg/ml) and in vivo (Extracts: 50, 100 and 200 mg/kg) antioxidant studies were performed on fresh goat liver and rats (N = 45) of either sex, respectively. In vitro hepatoprotective efficacy of extracts was evaluated against ethanol induced toxicity in HepG2 cell line. In vivo study was performed at 50, 100 and 200 mg/kg/day doses in rats by CCl4-induced hepatotoxicity study. ResultsNo mortality was observed during single and repeated dose toxicity studies. 50% lethal dose >2000 mg/kg, confirmed category 5 toxicity level of extracts, according to Globally Harmonized System. No signs of toxicity and treatment or dose related changes recorded in rats under repeated dose toxicity study. No-observed-adverse effect-level of 200 mg/kg/day was observed for both extracts. Median lethal concentration of REE and RME were 1291.30 and 1045.89 μg/ml, respectively in BSL bioassay and 50% cytotoxicity concentration >1000 μg/ml was obtained for both extracts from MTT assay. Calculated 50% inhibitory concentration and median effective dose of extracts obtained from different antioxidant assays in ex vivo and in vivo antioxidant studies, respectively indicated REE has more antioxidant efficacy than RME. In in vitro hepatoprotective efficacy study, extracts demonstrated dose dependent protection against ethanol induced hepatotoxicity. At 400 μg/ml, REE and RME demonstrated percentage protection of 65.53% and 57.98%, respectively.Results of liver function test and histopathological evaluation of liver in in vivo hepatoprotective study confirmed dose dependent protection provided by the extracts against CCl4 -induced hepatotoxicity. ConclusionBoth REE and RME were found safe to be considered for therapeutic uses. Both REE and RME were found to exhibit antioxidant efficacy in ex vivo and in vivo models. Results ascertained that H. rostrata rhizomes possess significant hepatoprotective potency.

Moringa oleifera Lam. leaf extract mitigates carbon tetrachloride-mediated hepatic inflammation and apoptosis via targeting oxidative stress and toll-like receptor 4/nuclear factor kappa B pathway in mice

Carbon tetrachloride (CCl4) is a hepatotoxin that triggers liver damage. This study aimed to evaluate the protective effect of phytochemicals detected in Moringa oleifera Lam. leaf extract (MOLE) on CCl4-induced hepatotoxicity in mice. Phytochemicals, total phenolics, and total flavonoids were detected in MOLE. MOLE markedly decreased the elevation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in consistence with the ameliorating effect on CCl4-induced histopathological abnormalities. Moreover, MOLE significantly alleviated the decrease in the antioxidant defense mechanism induced by CCl4. The suppressing effect of MOLE on the boosted inflammatory pathway triggered by CCl4 was detected by measuring the protein levels of nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-κB-p65), toll-like receptor 4 (TLR4), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β, and IL-8 as well as the relative expressions of nuclear factor kappa B (NF-κB), TNF-α, IL-1β, and TLR4 genes. Apoptosis and genotoxicity induced by CCl4 were significantly alleviated by MOLE. MOLE co-administration modulated TLR4/NF-κB pathway as presented by the suppressed gene expression of TLR4 and NF-κB as well as by the reduced protein expression of TLR4 and NF-κB-p65. In conclusion, MOLE has a multifarious protective role against hepatotoxicity through control of oxidative stress and modulation of TLR4/NF-κB.

Study The protective Effects of Cichorium intybus L. Root Extract Against Carbon Tetrachloride-Induced Hepatotoxicity in Broiler Chickens

An existing survey was performed to assess the hepatoprotective effects of C. intybus root extract on carbon tetrachloride-induced hepatotoxicity in broiler chickens. A total of 240 one-day-old broiler chickens were divided into diverse groups of control, carbon tetrachloride (1 mL/kg body weight IM injection), C. intybus root extract (500 mg/kg body weight/daily orally) and finally both C. intybus root extract and carbon tetrachloride. Liver samples were dissected and examined histopathologically at the day 31 of maintenance. Numerical grading of histopathological findings was performed according to the types of lesions at different criteria. Liver samples of carbon tetrachloride-treated group harbored moderate fatty degeneration and necrosis. Those treated with C. intybus extract showed normal cellular and tissue structure. Chicks faced with carbon tetrachloride and treated with C. intybus extract exhibited mild fatty degeneration and normal tissue structure. Chicks treated with C. intybus extract displayed the lowermost scores of necrosis (1.08±0.28), degeneration (1.33±0.49), inflammation (1.16±0.38) and total pathologic indexes (3.58±0.79). Broilers faced with carbon tetrachloride displayed the uppermost scores of necrosis (2.83±0.57), degeneration (3.66±0.49), inflammation (2.16±0.38) and total pathologic indexes (8.66±0.98). C. intybus extract application to broilers faced with carbon tetrachloride caused statistically significant decrease in the scores give to necrosis, degeneration, inflammation and total pathologic indexes (P <0.05). Findings suggested that 10 days administration of C. intybus extract caused significant decrease in the formation of centrilobular hepatic necrosis, fatty changes, inflammation and degeneration of the hepatic cells of broilers faced with CCl4-induced hepatotoxicity.

Structural characterization and amelioration of sulfated polysaccharides from Ganoderma applanatum residue against CCl4-induced hepatotoxicity

Natural polysaccharides and their derivatives have attracted academic attention due to their extensive physiological activities. However, the hepatoprotective effects against carbon tetrachloride (CCl4) toxicity have not been well elucidated. The objectives of this study were to characterize the structural properties of sulfated Ganoderma applanatum residue polysaccharides (SGRP) and to evaluate their inhibitory effects on liver fibrosis caused by oxidative stress and inflammation. Our in vivo study showed that SGRP was hepatoprotective in CCl4-induced chronic liver injury mice. It reduced the histopathological damages, down-regulated CYP2E1 (cytochrome P450 2E1) expression, reduced serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, improved the anti-oxidative and anti-inflammatory properties, inhibited TLR4/NF-κB signaling pathway, and reduced the release of inflammatory cytokines. The structural studies indicated that SGRP is a heteropolysaccharide with 7.8% sulfur content and α-linked residue. Our study projects SGRP as a potential candidate in anti-fibrosis treatment by using it as a food supplement or in medicines produced by pharmaceutical industries.

GC/MS-Based Metabolomics Profiling Approach and Determination of Ameliorative effect of Chiliadenus montanus extract towards CCl4 induced Hepatotoxicity in Albino Rats

The current study aimed to evaluate the GC/MS profiling of Chiliadenus montanus leaves extract which consists of 48 compounds, nine of them are major and represented 52.55 % from the total peak areas. The ameliorative effect towards the CCl4-induced hepatotoxicity in male Wistar rats through measuring certain biochemical parameters content in the liver were analyzed. CCl4 diluted 1:9 (v/v) in olive oil was injected intraperitoneally followed by oral administration of methanolic plant extract (200 mg/kg body weight). The CCl4-treated rats showed a significant decline in the studied hematological parameters, the serum levels of high-density lipoprotein (HDL), albumin (A) as well as the hepatic levels of glutathione (GSH) and activities of catalase (CAT), superoxide dismutase (SOD) , glutathione reductase (GR), elevation in the levels of total lipids (TL), triglycerides (TG), total cholesterol (TC), low-density lipoproteins (LDL), globulin (G), total bilirubin (TBil) , alanine and aspartate aminotransferase and alkaline phosphatase (ALAT and ASAT, ALP) and the hepatic levels of malondialdehyde (MDA). In contrast, the administration of methanol extract, notably improved all the studied parameters. This study showed that CCl4 administration to Wistar rats, at a high dose level, could induce a hepatic injury in addition to certain hematologic and metabolic alterations. The work was extended to examine tissue histopathology. In conclusion, C. montanus leaves methanol extract, resulted in an attractive candidate for ameliorating of hepatotoxicity induced by CCl4 through scavenging free radicals, improved liver functions, and normalizing the liver histopathological architecture. Yet, the treatment with methanol extract could ameliorate these alterations via their antioxidative effect. Further studies are required in order to identify the molecules responsible of the pharmacological activities.

A study to evaluate the hepatoprotective activity of prebiotics, probiotics, and synbiotic in CCl4 induced hepatotoxicity in rats

The current study was evaluated for the hepatoprotective activity of probiotics, prebiotics alone, and synbiotic of them. Animals were grouped into 12 groups (n = 6), i.e. normal control (NC), disease control (DC), treatment with prebiotics, probiotics alone, and synbiotic of them, and standard marketed hepatoprotective Silymarin along with Synbiotic 2000. All treatments were orally administered daily for 28 days. On the 28th day, all animals except the NC group received carbon tetrachloride (CCl4) (1 ml/kg, i.p.). Levels of serum glutamic pyruvic transaminase (SGPT), serum glutamic-oxaloacetic transaminase (SGOT), alkaline phosphatase (ALP), total bilirubin, total protein, lactate dehydrogenase, and urea were evaluated. The tissues were evaluated for oxidative markers levels of catalase (CAT), lipid peroxidation (LPO), and glutathione (GSH), and total protein. Histopathology of tissues was carried out. Compositional analysis of intestinal microbiota was done by 16S amplicon sequencing. CCl4 caused elevation in SGPT, SGOT, ALP, total bilirubin, urea, and lactate dehydrogenase in the DC group which was prevented with pretreatment of prebiotics, probiotics, and synbiotics. Total protein was decreased in serum and tissues after administration of CCl4 which was restored with pretreatment of test substances. Oxidative marker LPO was increased in the DC group and catalase (CAT) and GSH were also depleted in liver tissue. These changes were prevented in pretreated test groups. The histological change in hepatic parenchyma and hepatocytes was minimal in test groups compared to DC. The 16S amplicon sequencing indicated that Lactobacillus acidophilus and prebiotic treatment effectively displaced Staphylococcus. Results suggested that the use of prebiotics, probiotics alone, and synbiotic of them has a protective effect against CCl4-induced hepatotoxicity in rats.

Attenuation of liver mitochondrial oxidative damage by the extract and desulfo glucosinolate fraction of Lepidium perfoliatum L. seeds

Glucosinolates are the main bioactive compounds present in Brassicaceae family with remarkable protective activities against various chemical carcinogenesis and toxicities. Protective role of the hydroalcoholic extract and desulfo glucosinolate fraction from seeds of Lepidium perfoliatum were evaluated against CCl4-induced hepatotoxicity in BALB/c mice by determination of oxidative stress biomarkers and liver histopathological feature. The oral administration of different doses of the extract and desulfo glucosinolate fraction exhibited significant decrease in ROS and MDA contents of mitochondria in a dose-dependent manner, especially the doses of 200 mg/kg of the extract and 10 mg/kg of the fraction. GSH content was significantly increased (p<0.05) only in the group fed with 10 mg/kg of the fraction. Treatment groups showed notable reduction in ALT, AST and ALP levels. Based on the histopathological findings, the destructive effects of CCl4 were alleviated by the treated groups however the fraction (10 mg/kg) showed the best results. Based on NMR and EI-MS spectra, an alkyl glucosinolate bearing a thiofunction named glucoerucin was identified as the major constituent (80%) of the fraction. In addition, the GC-MS analysis revealed the presence of glucoerucin hydrolysis products, erucin and erucin nitrile, which representing 97.6% of the total seed volatiles. Based on the present study, it seems that glucoerucin is involved in the protection of L. perfoliatum against CCl4-induced hepatic damage via the inhibition of oxidative damage pathways.

Green coffee methanolic extract and silymarin protect against CCl4-induced hepatotoxicity in albino male rats

BackgroundDuring the last few decades, patients worldwide have been interested in using alternative medicine in treating diseases to avoid the increased side effects of chemical medications. Green coffee is unroasted coffee seeds that have higher amounts of chlorogenic acid compared to roasted coffee. Green coffee was successfully used to protect against obesity, Alzheimer disease, high blood pressure and bacterial infection.MethodsThis study aimed to investigate the probable protective activity of the green coffee methanolic extract, silymarin and their combination on CCl4-induced liver toxicity in male rats. Thirty Sprague – Dawley male albino rats were divided into 5 groups; control negative (G1) just got the vehicle (olive oil) and the other four groups received CCl4 dissolved in olive oil through an intraperitoneal injection and were divided into untreated control positive group (G2), the third group (G3) was treated with green coffee methanolic extract, the fourth group (G4) was treated with silymarin, and the fifth group (G5) was treated with a combination of green coffee methanolic extract and silymarin.ResultsIn the positive control group treated with CCl4 (G2), the CCl4-induced toxicity increased lipid peroxidation, IL-6, kidney function parameters, liver function enzymes, total cholesterol, triglycerides and low-density lipoproteins, and decreased irisin, antioxidants, CYP450 and high-density lipoprotein levels. Hepatic tissues were also injured. However, treating the injured rats in G3, G4 and G5 significantly improved the altered parameters and hepatic tissues.ConclusionsGreen coffee methanolic extract, silymarin, and their combination succeeded in protecting the male rats against CCl4 hepatotoxicity due to their antioxidant activity. Effect of green coffee methanolic extract mixed with silymarin in G5 was more efficient than that of green coffee methanolic extract in G3 or silymarin in G4.

Arctigenin attenuates CCl4-induced hepatotoxicity through suppressing matrix metalloproteinase-2 and oxidative stress

BackgroundIn spite of the huge advances in recent medicine, there is no effective drug that completely protects the liver from toxic materials. This study was conducted to investigate the hepatoprotective effect of arctigenin from burdock (Arctium lappa) against carbon tetrachloride (CCl4)-induced liver injury.ResultsArctigenin pre-administration reduced hepatotoxicity markers significantly as compared to CCl4 group. In addition, both silymarin and arctigenin declined matrix metalloproteinase-2 (MMP-2) in the serum (1177 ± 176), (978 ± 135) significantly as compared to CCl4 group (1734 ± 294). The hepatic antioxidant parameters (total glutathione, superoxide dismutase, and glutathione reductase) were significantly decreased after CCl4 injection, an effect that has been prevented by pre-administration of both silymarin and arctigenin. Histological examinations illustrated that arctigenin reduced CCl4 damage, where it decreased inflammation, congestion, and ballooning.ConclusionsArctigenin exerted a hepatoprotective effect against CCl4-induced liver damage in terms of suppressing MMP-2 and oxidative stress comparative to that of silymarin.

Hepatoprotective activity of hydroalcoholic extract of Cissampelos pareira linn. leaves against CCl4-induced acute and chronic hepatotoxicity

Background and objective Cissampelos pareira L. is a medicinal plant distributed across the tropics and used across the world traditionally for curing various pathological conditions. Hence, the present study has been carried out to evaluate the hepatoprotective effect of hydroalcoholic extract of C. pareira L. Materials and methods C. pareira L. leaves were extracted with a hydroalcoholic solvent. The resulting extract was subjected to an acute oral toxicity test on the basis of the OECD 423 guideline. Afterward, the selected dose of C. pareira hydroalcoholic extract (CPHE) was checked for hepatoprotective activity against CCl4-induced acute and chronic hepatotoxicity. Measurements of serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, total bilirubin, and direct bilirubin were performed. At the end of the study, histopathological analysis of livers of the animals of various treatment groups was carried out. Result and conclusion Based on the acute oral toxicity study, three doses of CPHE were selected, namely, 100, 200, and 400 mg/kg. Administration of CPHE at 200 and 400 mg/kg prevented an increase in serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, and bilirubin levels against CCl4-induced hepatotoxicity. The histopathological investigation of the portal triad structure of the liver clearly indicated that CPHE at 400 mg/kg showed significantly greater reduction in the necrotized area and normal appearance of the central vein, Kupffer cells and hepatocyte cells with no inflammatory cells. The results indicated that CPHE at 400 mg/kg protected the hepatic cells’ membrane integrity against CCl4-induced hepatotoxicity.

AMPK-mTOR-ULK1-mediated autophagy protects carbon tetrachloride-induced acute hepatic failure by inhibiting p21 in rats

Autophagy is a lysosomal-dependent degradation pathway in eukaryotic cells. Recent studies have reported that autophagy can facilitate the activation of hepatic stellate cells (HSCs) and fibrogenesis of the liver during long-term carbon tetrachloride (CCl4) exposure. However, little is known about the role of autophagy in CCl4-induced acute hepatic failure (AHF). This study aimed to identify whether modulation of autophagy can affect CCl4-induced AHF and evaluate the upstream signaling pathways mediated by CCl4-induced autophagy in rats. The accumulation of specific punctate distribution of endogenous LC3-II, increased expression of LC3-II, Atg5, and Atg7 genes/proteins, and decreased expression of p62 gene were observed after acute liver injury was induced by CCl4 in rats, indicating that CCl4 resulted in a high level of autophagy. Moreover, loss of autophagic function by using chloroquine (CQ, an autophagic inhibitor) aggravated liver function, leading to increased expression of p21 (a cyclin-dependent kinase inhibitor) in CCl4-treated rats. Furthermore, the AMPK-mTORC1-ULK1 axis was found to serve a function in CCl4-induced autophagy. These results reveal that AMPK-mTORC1-ULK1 signaling-induced autophagy has a protective role in CCl4-induced hepatotoxicity by inhibiting the p21 pathway. This study suggests a useful strategy aimed at ameliorating CCl4-induced acute hepatotoxicity by autophagy.

Hepatoprotective effects of Tagetes lucida root extract in carbon tetrachloride-induced hepatotoxicity in Wistar albino rats through amelioration of oxidative stress

Context The roots of Tagetes lucida Cav. (Asteraceae) have antioxidant and antimicrobial properties. Objective This study aimed to examine the hepatoprotective effects of T. lucida roots ethanol extract (TLRE) using carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. Materials and methods The active ingredients of TLRE were identified by high-performance liquid chromatography, infra-red spectrum, and mass spectrometric procedures. Ninety rats were distributed into four main groups: positive, therapeutic, protective, and negative group. The therapeutic group was implemented using CCl4 (a single dose of 2 mL/kg) before TLRE or silymarin administration. Meanwhile, the protective group was implemented by administering CCl4 (a single dose of 2 mL/kg) after force-feeding TLRE or silymarin. Each therapeutic and protective group was divided into three subgroups: force-fed with saline, TLRE (500 mg/kg), and silymarin (25 mg/kg). The positive group was split into two subgroups that were force-fed TLRE and silymarin. Positive, therapeutic, and protective groups were compared to the negative group (untreated rats). CCl4, TLRE, and silymarin were orally administrated using a gastric tube. Results In the therapeutic and protective groups, TLRE significantly reduced liver enzymes, i.e., aspartate aminotransferase (12.47 and 6.29%), alanine aminotransferase (30.48 and 11.39%), alkaline phosphatase (17.28 and 15.90%), and cytochrome P450-2E1 (39.04 and 48.24%), and tumour necrosis factor-α (53.72 and 53.72%) in comparison with CCl4-induced hepatotoxicity controls. Conclusions TLRE has a potent hepatoprotective effect with a good safety margin. After a repeated study on another type of small experimental animal, their offspring, and an experiment with a large animal, this study may lead to clinical trials.