A novel solid dispersion system for natural product-loaded medicine: silymarin-loaded solid dispersion with enhanced oral bioavailability and hepatoprotective activity

Abstract

A surface-attached silymarin-loaded solid dispersion with improved dissolution profile and enhanced oral bioavailability was formulated using silymarin, polyvinylpyrrolidone (PVP) and Tween 80 in water. In this solid dispersion, hydrophilic PVP was adhered onto the surface of crystalline drug rendering silymarin hydrophilic without changing its crystallinity. The drug solubility from the optimised solid dispersion prepared with silymarin/PVP/Tween 80 at the weight ratio of 5/2.5/2.5 increased by almost 650-fold compared to drug powder. The drug was physically and chemically stable in the solid dispersion for at least 6 months. Moreover, the solid dispersion enhanced the oral bioavailability of the drug in rats by almost 3-fold compared to the commercial product. The silymarin-loaded solid dispersion also exhibited advanced hepatoprotective bioactivity against CCl₄-induced liver damage compared to silymarin or the commercial product. Thus, this silymarin-loaded solid dispersion would be useful for the enhancement of oral bioavailability and hepatoprotective activity of poorly water-soluble silymarin.     

Protective effect of ethyl acetate fraction of Rhododendron arboreum flowers against carbon tetrachloride-induced hepatotoxicity in experimental models

Objective:

To evaluate the hepatoprotective potential of ethyl acetate fraction of Rhododendron arboreum (Family: Ericaceae) in Wistar rats against carbon tetrachloride (CCl₄)-induced liver damage in preventive and curative models.

Materials and Methods:

Fraction at a dose of 100, 200, and 400 mg/kg was administered orally once daily for 14 days in CCl₄-treated groups (II, III, IV, V and VI). The serum levels of glutamic oxaloacetic transaminase (SGOT), glutamate pyruvate transaminase (SGPT), alkaline phosphatase (SALP), γ-glutamyltransferase (γ -GT), and bilirubin were estimated along with activities of glutathione S-transferase (GST), glutathione reductase, hepatic malondialdehyde formation, and glutathione content.

Result and Discussion:

The substantially elevated serum enzymatic activities of SGOT, SGPT, SALP, γ-GT, and bilirubin due to CCl₄ treatment were restored toward normal in a dose-dependent manner. Meanwhile, the decreased activities of GST and glutathione reductase were also restored toward normal. In addition, ethyl acetate fraction also significantly prevented the elevation of hepatic malondialdehyde formation and depletion of reduced glutathione content in the liver of CCl₄-intoxicated rats in a dose-dependent manner. Silymarin used as standard reference also exhibited significant hepatoprotective activity on post-treatment against CCl₄-induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that ethyl acetate fraction has a potent hepatoprotective action against CCl₄-induced hepatic damage in rats.     

Hepatoprotective effects of the nitric oxide donor isosorbide-5-mononitrate alone and in combination with the natural hepatoprotectant, silymarin, on carbon tetrachloride-induced hepatic injury in rats

The aim of this study was to investigate the effect of the nitric oxide donor isosorbide-5-mononitrate (5-ISMN) alone or in combination with the natural hepatoprotectant with anti-oxidant activity silymarin on the carbon tetrachloride (CCl₄)-induced hepatic injury in rats. 5-ISMN (1.8, 3.6 or 7.2 mg/kg), silymarin (25 mg/kg) or 5-ISMN (1.8, 3.6 or 7.2 mg/kg) combined with silymarin was given once daily orally simultaneously with CCl₄ and for 15 days thereafter. Liver damage was assessed by determining serum enzyme activities and hepatic histopathology. 5-ISMN given at the above doses conferred significant protection against the hepatotoxic actions of CCl₄ in rats, reducing serum alanine aminotransferase (ALT) levels by 31.2, 39.3 and 61.6%, respectively, when compared with controls. Serum aspartate aminotransferase (AST) levels decreased by 19.8, 22.7 and 59.4%, respectively, while alkaline phosphatase (ALP) decreased by 26.1 and 32.6% by the drug at 3.6 and 7.2 mg/kg, respectively. When silymarin was added to 5-ISMN (1.8, 3.6 or 7.2 mg/kg), ALT decreased by 32.8, 59.6, 70.2% and AST by 28.7, 50.3, 60%, when compared with CCl₄ control group levels. Silymarin in combination with 3.6 or 7.2 mg/kg 5-ISMN resulted in 37.5 and 39.2% reductions in ALP when compared with CCl₄ control group. Meanwhile, silymarin alone reduced ALT, AST and ALP levels by 65.9, 52 and 62.3%, respectively. Blood levels of reduced glutathione were markedly decreased in CCl₄-treated rats. Reduced glutathione levels were increased by the administration of 5-ISMN and restored to near normal values by silymarin treatment. Histopathological alterations by CCl₄ were markedly reduced after treatment with 5-ISMN alone or in combination with silymarin. Histopathologic examination of the livers of CCl₄-treated rats administered 5-ISMN at 7.2 mg/kg showed marked restoration of the normal architecture of the liver tissue and minimal fibrosis. Silymarin co-administered with 5-ISMN resulted in further improvement of the histologic picture. These results indicates that treatment with 5-ISMN protects against hepatocellular necrosis induced by CCl₄. The study suggests a potential therapeutic use for 5-ISMN in combination with silymarin in liver injury.     

Hepatoprotective activity of punarnavashtak kwath,an Ayurvedic formulation,against CCl4-induced hepatotoxicity in rats and on the HepG2 cell line

Objective: Punarnavashtak kwath (PNK) is a classical Ayurvedic formulation, mentioned in Ayurvedic literature Bhaishajya Ratnavali, for hepatic disorders and asthma. This study investigated the hepatoprotective activity of PNK to validate the traditional use of this formulation.

Materials and methods: PNK was prepared in the laboratory according to the method given in Ayurvedic literature. Phytochemical screening was performed to determine the presence of phytoconstituents. Hepatoprotective activity was evaluated against CCl₄-induced hepatotoxicity in rats and by its effect on the HepG2 cell line.

Results: Preliminary phytochemical screening revealed the presence of alkaloids, tannins, flavonoids, saponins, and a bitter principle in PNK. Administration of PNK produced significant hepatoprotective effect as demonstrated by decreased levels of serum liver marker enzymes such as aspartate transaminase, serum alanine transaminase, serum alkaline phosphatase, and serum bilirubin and an increase in protein level. Thiopentone-induced sleeping time was also decreased in the PNK-treated animals compared with the CCl₄-treated group. It also showed antioxidant activity by increase in activity of glutathione, superoxide dismutase, and catalase and by a decrease in thiobarbituric acid reactive substance level compared with the CCl₄-treated group. Results of a histopathological study also support the hepatoprotective activity of PNK. Investigation carried out on the HepG2 cell line depicted significant increase in viability of cells exposed to PNK as compared with CCl₄-treated cells.

Discussion and Conclusion: It can be concluded that PNK protects hepatocytes from CCl₄-induced liver damages due to its antioxidant effect on hepatocytes. An in vitro study on HepG2 cell lines also supports its protective effect.     

Protective effects of silica hydride against carbon tetrachloride-induced hepatotoxicity in mice

The protective effects of MegaHydrate™ silica hydride against liver damage were evaluated by its attenuation of carbon tetrachloride (CCl₄)-induced hepatotoxicity in mice. Male ICR mice were orally treated with silica hydride (104, 208 and 520 mg/kg) or silymarin (200 mg/kg) daily, with administration of CCl₄ (1 mL/kg, 20% CCl4 in olive oil) twice a week for eight weeks. The results showed that oral administration of silica hydride significantly reduced the elevated serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), triglyceride (TG), and cholesterol and the level of malondialdehyde (MDA) in the liver that were induced by CCl₄ in mice. Moreover, the silica-hydride treatment was also found to significantly increase the activities of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH-Px), as well as increase the GSH content, in the liver. Liver histopathology also showed that silica hydride reduced the incidence of liver lesions induced by CCl₄. The results suggest that silica hydride exhibits potent hepatoprotective effects on CCl₄-induced liver damage in mice, likely due to both the increase of antioxidant-defense system activity and the inhibition of lipid peroxidation.     

Antihepatotoxic activity of ferolactone, a new furanocoumarin from Feronia limonia

The Fruit of Feronia limonia Swingle (Rutaceae) afforded a new furanocoumarin, characterized as Monoterpenoid-5-methoxy furanocoumarin lactone, named as ferolactone. Ferolactone exhibited a significant antihepatotoxic activity by reducing the elevated levels of serum enzymes such as serum glutamate oxaloacetate transaminase (SGOT) by 43.44%, serum glutamate pyruvate oxaloacetate transaminase (SGPT) by 39.63%, and alkaline phosphatase (ALP) by 41.16%; while the total protein (TP) levels were increased by 36.15%. When compared with standard drug, silymarin decreased SGOT by 45.80%, SGPT by 43.67%, ALP by 37.96%, and increased TP levels by 55.16%, against CCl₄-induced toxicity in albino wistar rats. These biochemical observations were also supplemented by histopathological examinations of the liver sections, which showed significant recovery of hepatocytes of the liver in ferolactone-treated animals.     

Hepatoprotective effect of electrolyzed reduced water against carbon tetrachloride-induced liver damage in mice

The study investigated the protective effect of electrolyzed reduced water (ERW) against carbon tetrachloride (CCl₄)-induced liver damage. Male ICR mice were randomly divided into control, CCl₄, CCl₄ + silymarin, and CCl₄ + ERW groups. CCl₄-induced liver lesions include leukocytes infiltration, hepatocyte necrosis, ballooning degeneration, mitosis, calcification, fibrosis and an increase of serum alanine aminotransferase (ALT), and aminotransferase (AST) activity. In addition, CCl₄ also significantly decreased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). By contrast, ERW or silymarin supplement significantly ameliorated the CCl₄-induced liver lesions, lowered the serum levels of hepatic enzyme markers (ALT and AST) and increased the activities of SOD, catalase, and GSH-Px in liver. Therefore, the results of this study show that ERW can be proposed to protect the liver against CCl₄-induced oxidative damage in mice, and the hepatoprotective effect might be correlated with its antioxidant and free radical scavenging effect.     

Antihepatotoxic activity of debelalactone,a new oxirano-furanocoumarin from Phyllanthus debilis

The whole plant of Phyllanthus debilis (Euphorbiacae) afforded a new oxirano-furanocoumarin, characterized as 5-hydroxy-7-methoxy-furanocoumarin-9(14)-cyclohex-12(13)-oxirano-11-one (1), named as debelalactone. Debelalactone exhibited a significant antihepatotoxic activity by reducing the elevated levels of serum enzymes such as serum glutamate oxaloacetate transaminase (SGOT) by 59.14%, serum glutamate pyruvate oxaloacetate transaminase (SGPT) by 61.84%, and alkaline phosphatase (ALP) by 85.93%; while the total protein (TP) levels were increased by 110.35%, when compared with standard drug silymarin that have decreased SGOT by 77.03%, SGPT by 69.67%, ALP by 93.18%, and increased TP levels by 100.48%, respectively, against CCl₄-induced toxicity in Wistar rats. These biochemical observations were also supplemented by histopathological examinations of the liver sections.     

Protective effect of Acacia confusa bark extract and its active compound gallic acid against carbon tetrachloride-induced chronic liver injury in rats

Acacia confusa Merr. (Leguminosae), a species native to Taiwan, is widely distributed on the hills and lowlands of Taiwan, and has been traditionally used as a medicine. The hepatoprotective effects of A. confusa bark extract (ACBE) and its active constituent gallic acid were evaluated against carbon tetrachloride (CCl₄)-induced hepatotoxicity in rats. CCl₄-induced hepatic pathological damage and significantly increased the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and malondialdehyde (MDA) in plasma, and cytochrome P4502E1 (CYP2E1) protein expression in hepatic samples, and decreased the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) in erythrocytes. Treatment with ACBE, gallic acid or silymarin could decrease significantly the AST, ALT, and MDA levels in plasma, and CYP2E1 expression in liver tissues, and increase the activities of SOD and GPX in erythrocyte when compared with CCl₄-treated group. Liver histopathology also showed that ACBE, gallic acid or silymarin could significantly reduce the incidence of liver lesions induced by CCl₄. These results suggested that the ACBE and gallic acid exhibit potent hepatoprotection against CCl₄-induced liver damages in rats, and the hepatoprotective effects of ACBE and gallic acid may be due to the modulation of antioxidant enzymes activities and inhibition of lipid peroxidation and CYP2E1 activation.     

Hepatoprotective effects of sesamol loaded solid lipid nanoparticles in carbon tetrachloride induced sub‐chronic hepatotoxicity in rats

Sesamol is a phenolic component of sesame seed oil, which has been established as an antioxidant and also possesses potential for hepatoprotection. However, its protective role in carbon tetrachloride (CCl₄) induced sub‐chronic hepatotoxicity has not been studied. Limited oral bioavailability (BA) and rapid elimination (as conjugates) in rats is reported for sesamol. Considering its significant antioxidant potential and compromised BA, we packaged sesamol into solid lipid nanoparticles (S‐SLNs) to enhance its hepatoprotective bioactivity. S‐SLNs prepared by microemulsification method were nearly spherical in shape with an average particle size of 120.30 nm and their oral administration at 8 mg/kg body weight (BW) showed significantly (p < 0.001) better hepatoprotection than free sesamol (FS) and a well established hepatoprotective antioxidant silymarin [SILY (25 mg/kg BW); p < 0.05) in CCl₄ induced sub‐chronic liver injury in rats. Evaluations were done in terms of histological changes in the liver tissue, liver injury markers (serum alanine aminotransferase, serum aspartate aminotransferase, and serum lactate dehydrogenase); oxidative stress markers (lipid peroxidation, superoxide dismutase, and reduced glutathione) and proinflammatory response marker (tumor necrosis factor‐alpha). © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 520–532, 2016.     

Hepatoprotective activity of Stereospermum suaveolens against CCl4-induced liver damage in albino rats

The present study aims to evaluate the hepatoprotective activity of Stereospermum suaveolens DC (Bignoniaceae). Hepatoprotective activity is studied by carbon tetrachloride (CCl₄)-induced liver damage in albino rats. The degree of protection in this activity has been measured by using biochemical parameters such as serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), total bilirubin, LDL-cholesterol and SOD, CAT, GSH, total thiols, NO, and lipid peroxidation in liver tissue homogenate. The results suggest that the methanol stem bark extract of Stereospermum suaveolens at the doses 125, 250, and 500?mg/kg and reference standard Liv-52 treated group produced significant (p <0.001) hepatoprotection against CCl₄-induced liver damage by decreasing the activities of serum enzymes, bilirubin and lipid peroxidation. The extract significantly (p <0.001) increased levels of SOD, CAT, GSH and total thiols, as compared to control group. Histopathological studies further substantiate the protective effect of the extract. It was concluded that methanol stem bark extract of Stereospermum suaveolens showed effective hepatoprotective activity.     

Protective Effect of Enicostemma littorale. against CCl4-Induced Hepatic Damage in Rats

Abstract

The alcohol extract of the whole plant of Enicostemma littorale. Blume was evaluated for its antihepatotoxic activity against CCl₄-induced hepatic damage in rats. The activity was evaluated by using biochemical parameters, such as serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, alkaline phosphatase, total bilirubin, and γ.-glutamate transpeptidase. The histopathological changes of liver sample were compared with respective controls. The extract showed a remarkable hepatoprotective effect.     

Mulberry water extracts (MWEs) ameliorated carbon tetrachloride-induced liver damages in rat

Mulberry extracts are antidiabetic and antihyperlipidemic, as well as preventive of cardiovascular disease. The current study investigates the protective mechanisms of mulberry water extracts (MWEs) in carbon tetrachloride (CCl₄)-induced hepatic injury. Oral administration of MWEs significantly reduced the lipid peroxidation triggered by CCl₄, as shown by the reduced production of thiobarituric acid-reactive substance (TBARS). The levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were also reduced via cotreatment with MWEs compared with CCl₄ treatment alone. Cotreatment with MWE evidently reduced CCl₄-induced liver weight and inhibited lipid deposition and fibrogenesis. In a similar manner, cotreatment with silymarin, a well-known liver protective agent, also reversed the CCl₄-induced effects, such as reduced TBARS formation, decreased serum AST, ALT, and ALP levels, blocked lipid accumulation, and liver fibrosis. Furthermore, MWEs attenuated the proinflammatory genes such as cyclooxygenase 2, nuclear factor kappa B, and inducible nitric oxide synthase expression. The current findings suggest that MWEs such as silymarin exhibit protective and curative effects against CCl₄-induced liver damage and fibrosis via decreased lipid peroxidation and inhibited proinflammatory gene expression.     

Protective effects of Pergularia daemia roots against paracetamol and carbon tetrachloride-induced hepatotoxicity in rats

Context: Pergularia daemia (Forsk) Chiov. (Asclepiadaceae) is a slender, hispid, fetid-smelling perennial climber and has been used for the treatment of inflammation, diabetes, malaria, asthma, and liver disorders. Ethnopharmacological surveys conducted among herbal practitioners of Western Ghats, Tamil Nadu, India, revealed that large numbers of laticiferous plant species are used as a source of herbal therapies, in which Pergularia daemia was commonly used to treat liver disease and jaundice.

Objective: The hepatoprotective effect of aqueous and ethanol extracts of Pergularia daemia roots by paracetamol and carbon tetrachloride (CCl₄)-induced liver damage in rats was studied.

Materials and methods: The aqueous (PdAE) and ethanol (PdEE) extracts of Pergularia daemia were studied for their hepatoprotective effects on paracetamol and CCl₄-induced liver damage on Wistar albino rats. The degree of protection was measured by physical changes (liver weight), biochemical (serum gultamic pyruvic transaminase, serum gultamic oxaloacetic transaminase, alkaline phosphatase, direct bilirubin, total bilirubin, cholesterol and decrease in protein), antioxidant enzymes (lipid peroxidation and glutathione levels), and histological changes.

Results: Pretreatment with PdAE and PdEE significantly prevented the physical, biochemical, antioxidant enzyme levels and histological changes induced by paracetamol and CCl₄ in the liver. The effects of PdAE and PdEE were comparable to that of the standard drug silymarin. The ethanol extract was found to exhibit greater hepatoprotective activity than the aqueous extract.

Discussion and conclusion: These results indicate that Pergularia daemia could be useful in preventing chemically induced acute liver injury. From this study it can be concluded that the aqueous and ethanol extracts of P. daemia possess significant hepatoprotective activity.     

Protective effects of seabuckthorn (Hippophae rhamnoides L.) seed oil against carbon tetrachloride-induced hepatotoxicity in mice

The present study examined the protective effects of seabuckthorn (Hippophae rhamnoides L., SBT) seed oil on carbon tetrachloride (CCl₄)-induced hepatic damage in male ICR mice. Our results showed that oral administration of SBT seed oil at doses of 0.26, 1.30, and 2.60 mg/kg for 8 weeks significantly reduced the elevated levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), triglyceride (TG), and cholesterol at least 13% in serum, and the level of malondialdehyde (MDA) in liver at least 22%, that was induced by CCl₄ (1 mL/kg) in mice. Moreover, the treatment of SBT seed oil was also found to significantly increase the activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Rd), and GSH content in liver up to 134%. Our study found that the optimal dose of SBT seed oil was 0.26 mg/kg, as the minimum amount exhibiting the greatest hepatoprotective effects on CCl₄-induced liver injury. Overall, the hepatoprotective effect of SBT seed oil at all tested doses was found to be comparable to that of silymarin (200 mg/kg) and have been supported by the evaluation of the liver histopathology in mice.     

Protection of the flavonoid fraction from Rosa laevigata Michx fruit against carbon tetrachloride-induced acute liver injury in mice

Protective effect of the total flavonoids (TFs) from Rosa laevigata Michx fruit against carbon tetrachloride (CCl₄)-induced hepatotoxicity in mice was investigated. Pretreatment with TFs significantly decreased CCl₄-induced elevation of serum aspartate transaminase (AST) and alanine transaminase (ALT) activities as well as the relative liver weight. Histopathological observation also revealed that TFs reduced the incidence of liver lesions and improved hepatocyte abnormality. Moreover, oral administration of TFs significantly enhanced antioxidant enzyme activities (superoxide dismutase, catalase and glutathione peroxidase), increased the content of glutathione and decreased the content of malondialdehyde. Further research indicated that TFs prevented the DNA fragmentation and mitochondrial ultrastructural alterations caused by CCl₄ based on TUNEL and transmission electron microscopy (TEM) assays. Moreover, pretreatment with TFs down-regulated the protein expressions of CYP2E1, iNOS, NF-κB, Bak and Caspase-3. Quantitative Real-time PCR assay suggested that TFs markedly decreased the levels of TNF-α, Fas/FasL and Bax gene expressions, and increased the level of Bcl-2. This is the first time to report the significant hepatoprotective effect of TFs from R. laevigata Michx fruit against CCl₄-induced liver injury in mice and the action should be through reducing oxidative stress and suppressing inflammation and apoptosis.     

Prevention of Carbon Tetrachloride-Induced Hepatotoxicity by the Ethanol Extract of Capparis moonii Fruits in Rats

The effect of the ethanol extract of Capparis moonii Hook. f. Thoms. (Capparidaceae) fruits was studied in carbon tetrachloride (CCl₄)-induced hepatotoxicity in rats. The hepatotoxicity was induced in rats with the administration of 1 : 1 (v/v) mixture of CCl₄ in olive oil at the dose of 1 ml/kg subcutaneously on day 7. The ethanol extract of C. moonii (200 mg/kg) and the standard drug silymarin (25 mg/kg) were given orally from day 1 to day 9. The extract of C. moonii produced significant (p &lt; 0.001) lowering of the elevated Serum glutamic oxaloacetic transaminace (SGOT) Serum glutamicpyraric transaminase (SGPT), alkaline phosphatase (ALP), and a rise of depleted total protein when compared with the toxic control. The results were comparable with the standard drug silymarin.     

Hepatoprotective effects of Gentiana asclepiadea L. extracts against carbon tetrachloride induced liver injury in rats

This study is an attempt to evaluate the hepatoprotective activity of Gentiana asclepiadea L. against carbon tetrachloride-induced liver injury in rats. Methanol extracts of aerial parts (GAA) and roots (GAR) of G. asclepiadea at doses of 100, 200, and 400 mg/kg b.w. were orally administered to Wistar rats once daily for 7 days before they were treated with CCl₄. The hepatoprotective activity of the extracts in this study was compared with the reference drug silymarin. In CCl₄ treated animals, GAA and GAR significantly decreased levels of serum transaminases, alkaline phosphatase and total bilirubin, and increased the level of total protein. Treatment with the extracts resulted in a significant increase in the levels of catalase, superoxide dismutase and reduced glutathione, accompanied with a marked reduction in the levels of malondialdehyde, as compared to CCl₄ treated group. The histopathological studies confirmed protective effects of extracts against CCl₄-induced liver injuries. No genotoxicity was observed in liver cells after GAA treatment, while GAR showed only slight genotoxic effects by comet assay. Phytochemical analysis revealed the presence of sweroside, swertiamarin and gentiopicrin in high concentrations in both extracts. It could be concluded that the use of G. asclepiadea extracts in the treatment of chemical-induced hepatotoxicity.     

Therapeutic potential of morin against liver fibrosis in rats: Modulation of oxidative stress,cytokine production and nuclear factor kappa B

Therapeutic potential of morin, a member of flavonoid family, against carbon tetrachloride (CCl₄)-induced liver fibrosis in rats was investigated and compared with that of silymarin. Results show that treatment with morin (30 mg/kg/day) revealed attenuation in liver index and serum biomarkers of liver function that were enhanced by chronic CCl₄ intoxication. Further, morin inhibited the elevated levels of malondialdehyde and nitric oxide and restored hepatic reduced glutathione to its normal level. The increased production of hepatic hydroxyproline content by CCl₄ was markedly decreased by administration of morin. In addition, treatment with morin significantly attenuated the inflammatory responses caused by CCl₄ as evident by the decreased hepatic tumor necrosis factor-alpha (TNF-α) level, immunohistochemical expressions of inducible nitric oxide synthase and nuclear factor kappa B. Collectively, this study indicates that morin possesses antifibrotic effect in the CCl₄ model of fibrosis via reducing oxidative stress, inflammatory responses and fibrogenic markers.