CCl4-intoxicated Rats Research Articles

Methanol extract of Nymphaea lotus ameliorates carbon tetrachloride-induced chronic liver injury in rats via inhibition of oxidative stress.

Nymphaea lotus (NL) is an aquatic perennial plant used traditionally in the management of various liver diseases. In this study, the protective effect of methanol extract of NL against carbon tetrachloride (CCl4)-induced chronic hepatotoxicity in rats was investigated. Male Wistar rats were assigned into six groups of five rats each. Group I received corn oil (0.5 mL p.o.) and served as control, group II received CCl4 (1 mL/kg i.p., 1:3 in corn oil), group III received NL (200 mg/kg), and groups IV, V, and VI received CCl4+NL (50, 100, and 200 mg/kg, respectively) for 6 weeks. Twenty-four hours after the last exposure, rats were bled and killed. The activities of alanine aminotransaminase (ALT), aspartate aminotransferase (AST), and levels of total bilirubin (TB) in the serum, thiobarbituric acid reactive substances (TBARS), superoxide dismutase, catalase, glutathione peroxidase (GPx) and glutathione (GSH) in the liver, and histopathology of the liver were determined using standard procedures. NL significantly (p<0.05) lowered the levels of ALT, AST, and TB and exhibited antioxidant potentials in rats exposed to CCl4 relative to the control values. Specifically, NL at 100 and 200 mg/kg significantly (p<0.05) increased CCl4-induced decrease in hepatic GSH and GPx and also decreased the level of hepatic TBARS in CCl4-intoxicated rats. Histopathological findings revealed cellular infiltration and fibrosis in rats that received CCl4 only, which were ameliorated in rats that received NL+CCl4. The data suggest that NL exhibited hepatoprotective effects in CCl4-intoxicated rats via antioxidative mechanism.

HEPATOPROTECTIVE EFFECT OF ALHAGI MAURORUM BOISS (LEGUMINOSAE) ALCOHOLIC EXTRACT AGAINST HEPATOTOXICITY IN RATS

Abstract Alhagi maurorum (camel thorn plant) is a promising medicinal plant due to the presence of flavonoids and phenolic compounds as major contents of its constituents. This study was aimed to investigate the hepatoprotective effect of oral administration of alcoholic extract of Alhagi maurorum against CCl4-induced liver injury in rats after 4 weeks of treatment on serum levels of total cholesterol (TC), triglycerides (TG), lipoprotein fraction, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP); total protein (TP),albumin, total bilirubin (TBil) and oxidative stress markers such as glutathione (GSH), Malondialdehyde (MDA), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) were determined. Histopathological examination of liver was also performed .Forty two adult male (Sprague-Dawley strain) weighting 200+5 g were divided into six equal groups as follows: group1: negative control group, group 2: positive control (CCl4) group injected subcutaneously by a single dose of CCL4 (2 ml/kg b.wt) at the last day of the experiment, group 3: rats treated with standard drug Silymarin (200 mg/kg b.wt) once daily for 4weeks prior a single subcutaneous injection of CCL4 (2 ml/kg b.wt.) and groups (4), (5) and (6) were orally administered Alhagi maurorum alcoholic extract at doses of (200, 400 and 600) mg/kg b.wt once daily for 4 weeks prior a single subcutaneous injection of CCL4 (2 ml/kg b.wt.) to induce experimental hepatotoxicity.The results showed that oral administration of Alhagi maurorum alcoholic extract in a concentrations of 600 mg/kg b.wt to rats for 4 weeks prior inducing hepatotoxicity by CCl4 significantly improved TC, TG, lipoprotein fractions, decreased the elevated serum levels of liver enzymes (AST, ALT and ALP), total bilirubin and increased serum total protein when compared to the control positive group. Oxidative stress markers (MDA, GSH, GPx, SOD and CAT) were significantly improved as compared to the control positive group. Histopathological examination of liver section of rats orally given Alhagi maurorum alcoholic extract prior inducing hepatotoxicity by CCl4 showed alleviation of histological degeneration changes in protected groups compared to control positive group. It is concluded that Alhagi maurorum alcoholic extract has high hypolipidemic, hepatoprotective effect and antioxidant effects in CCl4-intoxicated rats.

Hepatoprotective effects of allyl isothiocyanate against carbon tetrachloride-induced hepatotoxicity in rat

We evaluated the hepatoprotective activity of allyl isothiocyanate (AITC) against carbon tetrachloride (CCl4)-induced liver injury in rats. Sprague Dawley rats were orally administered AITC at doses of 5 (AITC 5) and 50 (AITC 50) mg/kg body weight once daily for 3 days, with or without intraperitoneal injection of CCl4. Serum chemistry was assessed for changes in alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The enzyme activities of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) were examined in liver tissues, while pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) mRNA expression were analyzed using real-time polymerase chain reaction. And heme oxygenase-1 (HO-1) and ionized calcium binding protein-1 (Iba-1) immunoreactivities were evaluated by Western blot analysis and immunohistochemistry, respectively. In serum chemistry, the oral administration of AITC itself did not affect the serum levels of ALT or AST, furthermore pretreatment with AITC 5 and AITC 50 significantly reduced the ALT and AST activity levels that were elevated in CCl4-intoxicated rats. In addition, AITC significantly suppressed the reduction of SOD and CAT, and the elevation of MDA, TNF-α mRNA expression, on the other hands, induced the expression of HO-1 compared with those of the vehicle-treated CCl4 group. The histopathological evaluation and Iba-1 immunoreactivity also supported the hepatoprotective effects of AITC against CCl4-induced liver injury. These results suggest that AITC ameliorates oxidative liver injury, possibly through reducing lipid peroxidation, enhancing antioxidant enzymes, and suppressing Kupffer cells and macrophages.

Hepatoprotective Effect of Wheat-Based Solid-State Fermented Antrodia cinnamomea in Carbon Tetrachloride-Induced Liver Injury in Rat.

Antrodia cinnamomea (A. cinnamomea) is an indigenous medical fungus in Taiwan and has multiple biological functions, including hepatoprotective and immune-modulatory effects. Currently, the commercially available A. cinnamomea are mainly liquid- and solid-state fermented A. cinnamomea. However, the hepatoprotective effect of solid-state fermented A. cinnamomea has never been reported. Here we evaluate the ability of air-dried, ground and non-extracted wheat-based solid-state fermented A. cinnamomea (WFAC) to protect against carbon tetrachloride (CCl4)-induced hepatic injury in vivo. The results showed that oral administration of WFAC dose dependently (180, 540 and 1080 mg/kg) ameliorated the increase in plasma aspartate aminotransferase and alanine aminotransferase levels caused by chronic repeated CCl4 intoxication in rats. WFAC significantly reduced the CCl4-induced increase in hepatic lipid peroxidation levels and hydroxyproline contents, as well as reducing the spleen weight and water content of the liver. WFAC also restored the hepatic soluble protein synthesis and plasma albumin concentration in CCl4-intoxicated rats, but it did not affect the activities of superoxide dismutase, catalase, or glutathione peroxidase. In addition, a hepatic morphological analysis showed that the hepatic fibrosis and necrosis induced by CCl4 were significantly ameliorated by WFAC. Furthermore, the body weights of control rats and WFAC-administered rats were not significantly different, and no adverse effects were observed in WFAC-administered rats. These results indicate that WFAC is a nontoxic hepatoprotective agent against chronic CCl4-induced hepatic injury.

Hepatoprotective effect of grape seed oil against carbon tetrachloride induced oxidative stress in liver of γ-irradiated rat

Carbon tetrachloride (CCl4) and ionizing radiation are well known environmental pollutants that generate free radicals and induce oxidative stress. The liver is the primary and major target organ responsible for the metabolism of drugs, toxic chemicals and affected by irradiation. This study investigated the effect of grape seed oil (GSO) on acute liver injury induced by carbon tetrachloride (CCl4) in γ-irradiated rats (7Gy). CCl4-intoxicated rats exhibited an elevation of ALT, AST activities, IL-6 and TNF-α level in the serum. Further, the levels of MDA, NO, NF-κB and the gene expression of CYP2E1, iNOS and Caspase-3 were increased, and SOD, CAT, GSH-Px, GST activities and GSH content were decreased. Furthermore, silent information regulator protein 1 (SIRT1) gene expression was markedly down-regulated. Additionally, alterations of the trace elements; copper, manganese, zinc and DNA fragmentation was observed in the hepatic tissues of the intoxicated group. These effects were augmented in CCl4-intoxicated-γ-irradiated rats. However, the administration of GSO ameliorated these parameters. GSO exhibit protective effects on CCl4 induced acute liver injury in γ-irradiated rats that could be attributed to its potent antioxidant, anti-inflammatory and anti-apoptotic activities. The induction of the antioxidant enzymes activities, down-regulation of the CYP2E1, iNOS, Caspase-3 and NF-κB expression, up-regulation of the trace elements concentration levels and activation of SIRT1 gene expression are responsible for the improvement of the antioxidant and anti-inflammatory status in the hepatic tissues and could be claimed to be the hepatoprotective mechanism of GSO.

Protective Effects of Silymarin, Alone or in Combination with Chlorogenic Acid and/or Melatonin, Against Carbon Tetrachloride-induced Hepatotoxicity.

Objective:The aim of this study was to evaluate the hepatoprotective effects of silymarin (SIL), alone and combined with chlorogenic acid (CA) and/or melatonin (ME), using a rat model of carbon tetrachloride (CCl4)-induced injury.Materials and Methods:Hepatotoxicity was induced by a single dose of CCl4 (1 ml/kg, IP). One day after, rats were received SIL (200 mg/kg) alone or in combination with CA (60 mg/kg) and/or ME (20 mg/kg) for 21 days.Results:SIL significantly decreased serum alanine aminotransferase, inflammatory cytokines, and vascular endothelial growth factor levels. Histological alterations, fibrogenesis, oxidative DNA damage, inflammatory mediators, and caspase-3 activity were significantly attenuated in SIL treated CCl4-intoxicated rats. On the other hand, cytochrome P450 2E1 activity showed a significant decrease in the liver of CCl4-intoxicated rats, an effect that was reversed following treatment with SIL. All beneficial effects of SIL were markedly potentiated when combined with CA and/or ME.Conclusions:These data indicate that SIL, alone and combined with CA and/or ME, protected the liver against CCl4-induced hepatotoxicity via attenuating inflammation, oxidative DNA damage, apoptosis, and fibrotic changes. The significantly intensified hepatoprotective effects of SIL when combined with both CA and ME suggest a possible synergism. These synergistic effects need to be further confirmed using detailed studies.SUMMARY Silymarin, chlorogenic acid and melatonin possess in vivo hepatoprotective activitySilymarin, chlorogenic acid and melatonin attenuate fibrogenesis, oxidative DNA damage, inflammation and apoptosisChlorogenic acid and melatonin enhance the hepatoprotective effect of silymarin. Abbreviations used: SIL: silymarin, CA: chlorogenic acid, ME: melatonin, CCl4: carbon tetrachloride, CYP2E1, cytochrome P450 2E1, ALT: alanine aminotransferase, IL-6: interleukin 6, IFN-γ: interferon gamma, VEGF: vascular endothelial growth factor, TNF-α: tumor necrosis factor alpha, CRP: C-reactive protein, 8-OxodG: 8-Oxo-2’-deoxyguanosine, TGF-B1: transforming growth factor beta 1, HSCs: hepatic stellate cells.

A New Octadecenoic Acid Derivative from Caesalpinia gilliesii Flowers with Potent Hepatoprotective Activity.

Background:Caesalpinia gilliesii Hook is an ornamental shrub with showy yellow flowers. It was used in folk medicine due to its contents of different classes of secondary metabolites. In our previous study, dichloromethane extract of C. gilliesii flowers showed a good antioxidant activity.Aim of the Study:Isolation and identification of bioactive hepatoprotective compounds from C. gilliesii flowers dichloromethane fraction.Materials and Methods:The hepatoprotective activity of dichloromethane fraction and isolated compounds were studied in CCl4-intoxicated rat liver slices by measuring liver injury markers (alanine aminotransferase, aspartate aminotransferase and glutathione [GSH]). All compounds were structurally elucidated on the basis of electron ionization-mass spectrometry, one- and two-dimensional nuclear magnetic resonance.Results:A new 12,13,16-trihydroxy-14(Z)-octadecenoic acid was identified in addition to the known β-sitosterol-3-O-butyl, daucosterol, isorhamnetin, isorhamnetin-3-O-rhamnoside, luteolin-7,4’-dimethyl ether, genistein-5-methyl ether, luteolin-7-O-rhamnoside, isovanillic acid, and p-methoxybenzoic acid. Dichloromethane fraction and isorhamnetin were able to significantly protect the liver against intoxication. Moreover, the dichloromethane fraction and the isolated phytosterols induced GSH above the normal level.Conclusion:The hepatoprotective activity of C. gilliesii may be attributed to its high content of phytosterols and phenolic compounds.SUMMARY Bioactive Hepatoprotective phytosterols and phenolics from chloroform extract of Caesalpinia gilliesii Abbreviations used: ALT: Alanine Aminotransferase; AST: Aspartate aminotransferase; GSH: Glutathione; SC50: Scavenging Capacity 50 (SC 50); COSY: Correlation spectroscopy; NMR: Nuclear Magnetic Resonance; CC: Column chromatography; EI-MS: Electron-impact mass spectrometry; HSQC: Heteronuclear single-quantum correlation.

Oligonol Ameliorates CCl₄-Induced Liver Injury in Rats via the NF-Kappa B and MAPK Signaling Pathways.

Oxidative stress is thought to be a key risk factor in the development of hepatic diseases. Blocking or retarding the reactions of oxidation and the inflammatory process by antioxidants could be a promising therapeutic intervention for prevention or treatment of liver injuries. Oligonol is a low molecular weight polyphenol containing catechin-type monomers and oligomers derived from lychee fruit. In this study, we investigated the anti-inflammatory effect of oligonol on carbon tetrachloride- (CCl4-) induced acute hepatic injury in rats. Oral administration of oligonol (10 or 50 mg/kg) reduced CCl4-induced abnormalities in liver histology and serum AST and serum ALT levels. Oligonol treatment attenuated the CCl4-induced production of inflammatory mediators, including TNF-α, IL-1β, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) mRNA levels. Western blot analysis showed that oligonol suppressed proinflammatory nuclear factor-kappa B (NF-κB) p65 activation, phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38 mitogen-activated protein kinases (MAPKs) as well as Akt. Oligonol exhibited strong antioxidative activity in vitro and in vivo, and hepatoprotective activity against t-butyl hydroperoxide-induced HepG2 cells. Taken together, oligonol showed antioxidative and anti-inflammatory effects in CCl4-intoxicated rats by inhibiting oxidative stress and NF-κB activation via blockade of the activation of upstream kinases including MAPKs and Akt.

Hepatoprotective effects of zingerone on carbon tetrachloride- and dimethylnitrosamine-induced liver injuries in rats.

In this study, we investigated the hepatoprotective and anti-fibrotic effects of zingerone, one of the active components of ginger, against carbon tetrachloride (CCl4)- and dimethylnitrosamine (DMN)-induced liver injuries in rats, respectively. Oral administration of zingerone (10mg/kg) reduced CCl4-induced abnormalities in liver histology, serum alanine aminotransferase and aspartate aminotransferase levels, and liver malondialdehyde levels. Zingerone treatment attenuated CCl4-induced increases in inflammatory mediators, including tumor necrosis factor-α, interleukin-1β, cyclooxygenase-2, and inducible nitric oxide synthase mRNA levels. Western blot analysis showed that zingerone suppressed activation of nuclear factor-kappa B (NF-κB) p65 and phosphorylation of extracellular signal-regulated kinase, c-Jun NH2-terminal kinase, and p38 mitogen-activated protein kinases (MAPKs). Liver fibrosis induced by DMN (10mg/kg, intraperitoneally) was ameliorated by administration of zingerone (10 and 20mg/kg, orally). Zingerone treatment reduced DMN-induced elevation of hydroxyproline content and hepatic stellate cell activation. In conclusion, zingerone showed antioxidative and anti-inflammatory effects in CCl4-intoxicated rats by inhibiting oxidative stress and NF-κB activation via blockade of the activation of upstream MAPKs. Moreover, zingerone had hepatoprotective and anti-fibrotic effects against DMN-induced liver injury suggesting its usefulness in the prevention of liver inflammation and the development of hepatic fibrosis.

Assessment of the Potential Role of Silymarin Alone or in Combination with Vitamin E and/ or Curcumin on the Carbon Tetrachloride Induced Liver Injury in Rat

The aim of this study was to investigate the effective role of silymarin either alone or in a combination with vitamin E and/or curcumin against the toxic impact of carbon tetrachloride (CCl4) induced liver injury The results revealed that administration of silymarin alone or in a combination with vitamin E and/or curcumin for 21 consecutive days, 24 h after CCl4 injection to rats, markedly ameliorated DNA damaged and apoptosis markers in rat livers, proinflammatory markers including tumor necrosis factor- α (TNF- α) and C-reactive protein (CRP ) in rat livers as well as interleukin 6 (IL-6), interferon gamma (IFN-γ) and vascular endothelial growth factor (VEGF) in rat sera. These treatments also could ameliorated the alteration in cytochrome P450 2E1 (CYP2E1) activity in livers of CCl4 intoxicated rats as well as the increase in the serum alanine aminotransferase (ALT) compared with CCl4 intoxicated untreated rats. The present biochemical results are supported by histo-pathological examination. In conclusion, silymarin in a combination with vitamin E and curcumin was the most effective treatment in alleviating CCl4 induced liver damage and this may support the use of this combination as an effective treatment against liver damage induced by toxic agents.

The Effect of Electro-acupuncture at ST36 on Carbon Tetrachloride-induced Chronic Hepatic Damage in Rats

Objectives : The purpose of this study is to investigate the effect of electro-acupuncture(EA) at ST36 on CCl4 - induced chronic hepatic damage in rats. Methods : The authors performed several experimental items, including measurements of liver index, hematological analysis for WBC, lymphocytes, neutrophils, monocytes, hemoglobin, and biochemical assays for ALT, AST, ALP in serum, and histological analysis of liver tissue. Results : Liver index(liver weight/body weight), the levels of WBC, Neutrophils and Monocytes in blood, the levels of ALT, AST and ALP in serum which were increased by CCl4 - intoxication were reduced significantly by EA at ST36. The levels of lymphocyte and hemoglobin in blood which were decreased by CCl4 - intoxication were significantly increased by EA at ST36. Conclusions : EA at ST36 has a hepatoprotective effect on chronic hepatic damage in CCl4 intoxicated rats.

Potential therapeutic impacts of phyllanthus and costus aqueous extracts on CcL4 intoxicated rats

Bakground: Chronic liver damage is a worldwide common pathology, characterised by an inflammatory and fibrotic process that leads to a progressive evolution from chronic hepatitis to cirrhosis and hepatocellular carcinoma. A major role of fats and oxidative stress has been recently demonstrated in the pathogenesis of liver diseases. It was a newly introduced plant from Mexico. In India, it is grown in gardens, especially in Kerala, where the fresh leaves are eaten raw by diabetic people Objective: This work aimed to study the effect of phyllanthus and costus aqueous extracts on CcL4 intoxicated rats. Material and methods: Thiry mature male albino rats weighing 150±5g were obtained from Laboratory of Animal Colony, Helwan, Egypt. Rats were divided into 6 equal groups (n=5 rats each); one group was kept as a control negative, while the other 5 groups were injected subcutancosly (s/c) by carbon tetrachloride (CcL4) in parrafin oil 50% VN (2m1 / kg b.wt.) twice a week for two weeks for liver intoxication then , one group of them was left as control positive group (C +ve) while the rest four groups were given orally two doses (250 and 500 mg/kg) of phyllanthus and costus aqueous extracts (250 mg/kg and 500 mg/kg for the group. At the end of experimental period (45 days), blood samples were collected for serum separation to determine serum liver enzymes (AST, ALT and ALP), total cholesterol, triglycerides, atherosclerosis index, lipoprotein fraction (HDLc, LDLc and VLDLc), kidney function(Creatinine, Urea and Uric acid) and histopathological examination for the liver. Results: The obtained results demonstrated that phyllanthus and costus aqueous extracts caused significant improvement in liver enzymes, lipid profiles and kidney functions in CcL4 intoxicated rats. Conclusion: According to the results, phyllanthus and costus aqueous extracts could be tried on intoxicated patieats to improve liver &kidney function

Aqueous Date Flesh or Pits Extract Attenuates Liver Fibrosis via Suppression of Hepatic Stellate Cell Activation and Reduction of Inflammatory Cytokines, Transforming Growth Factor- β 1 and Angiogenic Markers in Carbon Tetrachloride-Intoxicated Rats.

Previous data indicated the protective effect of date fruit extract on oxidative damage in rat liver. However, the hepatoprotective effects via other mechanisms have not been investigated. This study was performed to evaluate the antifibrotic effect of date flesh extract (DFE) or date pits extract (DPE) via inactivation of hepatic stellate cells (HSCs), reducing the levels of inflammatory, fibrotic and angiogenic markers. Coffee was used as reference hepatoprotective agent. Liver fibrosis was induced by injection of CCl4 (0.4 mL/kg) three times weekly for 8 weeks. DFE, DPE (6 mL/kg), coffee (300 mg/kg), and combination of coffee + DFE and coffee + DPE were given to CCl4-intoxicated rats daily for 8 weeks. DFE, DPE, and their combination with coffee attenuated the elevated levels of inflammatory cytokines including tumor necrosis factor-α, interleukin-6, and interleukin-1β. The increased levels of transforming growth factor-β1 and collagen deposition in injured liver were alleviated by both extracts. CCl4-induced expression of α-smooth muscle actin was suppressed indicating HSCs inactivation. Increased angiogenesis was ameliorated as revealed by reduced levels and expression of vascular endothelial growth factor and CD31. We concluded that DFE or DPE could protect liver via different mechanisms. The combination of coffee with DFE or DPE may enhance its antifibrotic effects.

Appraisal of Total Phenol, Flavonoid Contents, and Antioxidant Potential of Folkloric Lannea coromandelica Using In Vitro and In Vivo Assays.

The aim of this study was to determine the impending antioxidant properties of different extracts of crude methanolic extract (CME) of leaves of Lannea coromandelica (L. coromandelica) and its two ethyl acetate (EAF) and aqueous (AqF) subfractions by employing various established in vitro systems and estimation of total phenolic and flavonoid content. The results showed that extract and fractions possessed strong antioxidant activity in vitro and among them, EAF had the strongest antioxidant activity. EAF was confirmed for its highest phenolic content, total flavonoid contents, and total antioxidant capacity. The EAF was found to show remarkable scavenging activity on 2,2-diphenylpicrylhydrazyl (DPPH) (EC50 63.9 ± 0.64 µg/mL), superoxide radical (EC50 8.2 ± 0.12 mg/mL), and Fe2+ chelating activity (EC50 6.2 ± 0.09 mg/mL). Based on our in vitro results, EAF was investigated for in vivo antioxidant assay. Intragastric administration of the EAF can significantly increase levels of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and glutathione peroxidase (GSH-Px) levels, and decrease malondialdehyde (MDA) content in the liver and kidney of CCl4-intoxicated rats. These new evidences show that L. coromandelica bared antioxidant activity.

Antioxidant and Hepatoprotective Activity of Fruit Extracts of Tetrapleura Tetraptera ( Schum and Thonn ) Taubert

The antioxidant and hepatoprotective activity of Tetrapleura tetraptera extracts in carbon tetrachloride (CCl 4)-induced liver injury in rats was investigated. T. tetraptera extracts showed varying levels of protective action against CCl 4-induced liver damage as evidenced through the significant reduction in the activities of serum marker enzymes for liver damage (alanine transaminase, aspartate transaminase, and alkaline phosphatase), and bilirubin levels when compared with CCl4intoxicated control rats. The extracts decreased the elevation in the activities of the enzymes in the liver. They also protected against CCl 4 induced lipid peroxidation. The extracts reduced CCl4-liver induced necrosis in dose dependent manner. These results indicated that fruit extracts of T. tetraptera possess a hepatoprotective property against CCl4induced liver damage which was mediated through its antioxidative defenses.

Mechanism of action involved in the hepatoprotective activities of methanol extract of Cassytha filiformis L. aerial parts in CCl4-induced liver damage

Cassytha filiformis alcohol decoction is used by traditional healers in Idem-Ogwa, Mbaitoli Local Government Area of Imo State, Nigeria, in the treatment of liver disorders ranging from hepatitis to susceptive poisoning and alcohol intoxication. The methanol extract of the aerial parts of Cassytha filiformis L. was investigated for hepatoprotective activity in rats challenged with carbon tetrachloride (CCl4). Apart from group 1 (normal), all other groups were pre-challenged with a single intraperitoneal injection of 0.7 ml of 20 % (v/v) CCl4 solution in olive oil. Group 2 (negative control) was given distilled water (10 ml kg−1, p.o.). Group 3 were treated with silymarin (35 mg kg−1 p.o.). Groups 4, 5, and 6 received oral treatments of 75, 150, and 300 mg kg−1 of the extract of C. filiformis respectively. All treatments were given by stomach intubation and at 12 h intervals for 7 days. Sodium pentobarbitone-induced sleeping time assay was carried out on the animals at 12 h after the last treatment on day 7 by intraperitoneal administration of Na-pentobarbitone (35 mg kg−1). Serum from individual animal was used to determine the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin, total protein and in vitro red blood cell membrane stability. The extract was safe up to 2000 mg kg−1 orally and did not induce overt clinical manifestation or death in rats. The extract significantly (p < 0.01) demonstrated potency at all test doses (75, 150, and 300 mg kg−1) in CCl4-challenged and treated animals when compared with prolonged Na-pentobarbitone-induced sleeping time observed in untreated CCl4-intoxicated rats. The methanol extracts of C. filiformis (150 and 300 mg kg−1) were also able to reduce hepatic tissue injury as revealed by a reduction in the serum levels of AST, ALT, ALP, and total bilirubin significantly (p < 0.05) but increased total serum protein significantly (p < 0.05) relative to values in untreated CCl4-induced liver damaged rats. Exposure of rat erythrocytes to varying saline concentrations (0.2–1.0 mg mL−1) of the extract did not produce observable measured increase in osmotic fragility in the red blood cell membranes. The findings suggest that the methanol extract of the aerial parts of C. filiformis could be useful in protecting hepatocytes from toxins especially from alcohol intoxication.

Investigation of the Antioxidant and Hepatoprotective Potential of Hypericum mysorense.

Background: Hypericum is a well-known plant genus in herbal medicine. Hypericum mysorense (Family: Hypericaceae), a plant belonging to the same genus, is well known in folklore medicine for its varied therapeutic potential. Objective: The aim of the present study was to investigate the different parts of the plant for antioxidant and hepatoprotective properties. Materials and Methods: The methanol extracts of Hypericum mysorense prepared from various parts of the plant were tested in vitro for their free radical scavenging activity against ABTS• (diammonium salt), DPPH• (1,1-diphenyl-2-picrylhydrazyl), NO•, O2•− and •OH radicals, using standard systems of assays. The total antioxidant capacity, total phenolic and total flavonoid content of the extracts were analyzed. Further, the leaf and flowering top extracts were tested for their in vivo antioxidant and hepatoprotective activities on Wistar rats using a carbon tetrachloride-induced hepatic injury model. Results: The leaf and flowering top extract showed potent antioxidant activity and also possessed highest total phenolic and flavonoid content. The antioxidant activity and the total phenolic and flavonoid content present in these extracts showed a good correlation. The leaf and flowering top extracts at 200 mg/kg restored aspartate amino transferase (ASAT), alanine amino transferase (ALAT), alkaline phosphatase (ALP), total bilirubin and protein levels significantly in CCl4-intoxicated rats. The tested extracts also showed a significant (p < 0.001) reduction in 2-thiobarbituric acid reactive substance (TBARS) levels with an increase in SOD and CAT levels. The histopathology of liver did not show any toxicity after the treatment with the extracts. The active extracts were standardized using two marker compounds, hyperoside and rutin, which were isolated from the plant by HPLC. HPLC studies revealed that the maximum concentration of hyperoside and rutin is present in the flowering top extract.

Fasting ameliorates metabolism, immunity, and oxidative stress in carbon tetrachloride-intoxicated rats

Fasting has been recently discovered to improve overall health, but its beneficial effects in the presence of hepatic insufficiency have not been proven. The influence of fasting on the metabolism, immunological aspects, and oxidative stress of 40 male carbon tetrachloride (CCl4)-intoxicated Wistar rats was investigated in the present study. The rats were divided into four groups, including a placebo group, CCl4-intoxicated rats, which were injected subcutaneously with 1.0 ml/kg of CCl4 solution, a fasting group, which was fasted 12 h/day for 30 days, and a fourth group, which was injected with CCl4 and fasted. The metabolism, immunity, and oxidative stress improved in CCl4-intoxicated rats fasted for 12 h/day for 30 days, as evidenced in significant increase (p < 0.05) in total protein, globulin, immunoglobulin M (IgM) and IgG levels, and total antioxidant capacity. In contrast, significant decrease (p < 0.05) in blood glucose, total cholesterol, low-density lipoprotein-cholesterol, alanine aminotransferase, C-reactive protein, and malondialdehyde levels were observed. Compared with CCl4-intoxicated rats, significant differences in the albumin, triacylglycerol, high-density lipoprotein-cholesterol, very low-density lipoprotein-cholesterol, cardiovascular risk factor, calcium and magnesium levels were not detected. The results of the present study showed that fasting improved metabolism, immunity, and oxidative stress in CCl4-intoxicated rats. Thus, fasting during Ramadan is safe for patients with hepatic disorders, as the prophet Mohammed (S) said "Keep the fast, keep your health".

Evaluation of the antioxidant activity of Marjoram plant (Origanium Majorana L.) in CCl 4-intoxicated rats.

Aim: This work was performed to evaluate the antioxidant activity and hepatoprotective effect of methanolic extract of Marjoram plant (Origanium Majorana L.) in CCl4- intoxicated rats. Materials and Methods: A total of 25 mature albino rats (Sprague Dawley) from both sexes, weighting from140 to 170 g were used. The animals were randomly divided into 5 groups each of 5 animals. These groups were used to investigate the in vivo hepatoprotective and antioxidant effects of the tested plant extract when given orally in two doses (375 and 750 mg/kg b.wt.). The first group was kept as normal healthy control. The other four groups were given CCl4 (1 ml/kg b.wt. s.c) for five days. One of these was used as a CCl4-intoxicated non treated control. Group III and IV were given the methanol extract of Origanium majorana plant orally at doses of 375 and 750 mg/kg b.wt. per day, for two months. Group V was used as a standard group and received silymarin orally at a dose of 100 mg/kg b.w. for two months. The antioxidant activity of methanol extract of Origanium majorana plant was studied in rats liver homogenate after the prolonged oral administration (2 months) via determination of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) enzyme activity. The in vitro antioxidant activity was compared to DPPH and measured spectrophotometrically. The activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in the different groups was evaluated as well as the histopathological examination of the liver. Results: Oral administration of Origanium majorana plant increased the antioxidant enzymes and significantly (P < 0.05) decreased liver enzyme levels when given in repeated doses. Conclusion: Thus methanolic extract of Origanium majorana possesses a significant antioxidant activity.

Hepatoprotective activity of Parmelia perlata (Huds.) Ach. against CCl4 induced liver toxicity in Albino Wistar rats

Article history: Many plants have been claimed to possess liver protecting activity. But, due to scarcity of potent modern medicine to treat severe liver diseases, many folk remedies of plant origin have been scientifically evaluated for their potential hepatoprotective activity in experimental animal models. Parmelia perlata (Huds.) Ach. (lichen, Parmeliaceae), commonly known as Dagad phool or stone flower in India, is mentioned in Indian Materia Medica as useful in treating a number of ailments. In the present study, hepatoprotective activity of aqueous slurry of P. perlata was investigated in CCl4 intoxicated Albino Wistar rats and the results were compared with, Silymarin. Intraperitoneal injection of CCl4, produced a marked elevation in the level of biochemical markers such as Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate Transaminase (SGPT), bilirubin, cholesterol and triglycerides. Oral administration of P. perlata aqueous slurry at 0.7 g / kg and 1.0 g / kg in CCl4 intoxicated rats showed marked decrease in the level of biochemical markers and results were at par with the effect shown by Silymarin. The results of histopathological analysis were in compliance with the findings of blood biochemical parameter analysis. This study confirms that P. perlata aqueous slurry has phyto- components with hepatoprotective potential.