CCl4-induced Hepatic Damage Research Articles

Objective: To evaluate the hepatoprotective effects of the ethanol fraction of Verbascum thapsus L. (EFVT) against CCl 4 -induced liver injury and elucidate its underlying mechanisms. Methods: The assessment of antioxidant properties and cell viability was conducted using the 2,2-diphenyl-1-picrylhydrazyl assay and HepG2 cells, respectively. The in vivo hepatoprotective efficacy of EFVT was evaluated in a rat model of carbon tetrachloride (CCl 4 )-induced liver injury by determining biochemical parameters, and oxidative stress- and inflammation-related markers. Gas chromatography-mass spectrometry was also employed for the qualitative analysis of its phytochemical composition. Results: GC-MS analysis of EFVT revealed the presence of several bioactive compounds such as 3 methyl mannoside and 9,12-octadecadienoic acid. Oral administration of EFVT significantly mitigated CCl 4 -induced liver injury, as evidenced by reduced levels of total bilirubin, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and malondialdehyde, boosted activities of catalase and superoxide dismutase, as well as enhanced glutathione levels. Histopathological examinations indicated EFVT restored abnormal liver architecture and reduced inflammation. Additionally, EFVT substantially downregulated the mRNA levels of IL-6, IL-1β, TNF-α , and NF-κB , and upregulated IL-10 expression. Conclusions: These findings underscore the therapeutic potential of EFVT in ameliorating liver damage associated with oxidative stress, providing scientific validation for its traditional utilization in ethnomedicine.

Hepatoprotective Effectiveness of Scutellaria linearis Benth Against CCl4-Induced Hepatic Damage in Mice

Background: This study investigated the antioxidant, hepatoprotective, and sedative modulatory effects of Nigella sativa alcoholic extract (NSAE) in CCl4-induced hepatotoxicity in rats. Methods: Male Wistar rats were divided into six groups (n=6): normal control, CCl4 control, silymarin (50 mg/kg), and NSAE (100, 200, and 400 mg/kg). Hepatoprotective effects were evaluated through biochemical parameters, oxidative stress markers, and histopathological examination. Results: NSAE treatment (400 mg/kg) significantly restored liver function markers, including SGOT (20.95 ± 0.52 IU/L, p = 0.033) and SGPT (28.61 ± 0.67 IU/L, p < 0.001), compared to CCl4 control. Total protein and albumin levels were normalized to 5.68 ± 0.54 mg/dL and 3.84 ± 0.48 mg/dL, respectively. Antioxidant parameters showed marked improvement with NSAE (400 mg/kg), increasing GSH (0.26 ± 0.029 µmol/mg) and CAT (30.19 ± 2.69 µg/mL) while reducing MDA (0.048 ± 0.008 µg/mL). Histopathological examination revealed significant protection against CCl4-induced hepatic and gastric tissue damage, particularly at the 400 mg/kg. Conclusion: NSAE exhibited marked hepatoprotective activity comparable to silymarin, predominantly through antioxidant mechanisms and the maintenance of hepatic tissue integrity, indicating its potential as a natural therapeutic agent for managing liver diseases. Because of its hepatoprotective and antioxidant properties, NSAE may be explored in clinical settings as a natural supplement to traditional liver disease therapies or as a prophylactic for people at risk of liver disorders.

Edgeworthia gardneri (Wall.) Meisn Mitigates CCL4-induced liver injury in mice by modulating gut microbiota, boosting antioxidant defense, and reducing inflammation.

Xeromphis nilotica is used by traditional practitioners in North East Nigeria to treat or manage various ailment such as inflammations, pains and stomach disorders. This study aimed to evaluate the probable protective effect of Xeromphis nilotica stem-bark extract against CCl4 induced hepatic injury in albino rats. Thirty-six rats were randomly divided into six groups. Group 1 served as the control and received 0.5mL of 1% tween 80 orally and 3ml/kgbwt olive oil intraperitoneally. Groups 2, 3, 4,5&6 were injected with 3ml/kgbwt of carbon tetra chloride (CCl4) in olive oil intraperitoneally to induce oxidative stress and thereafter group 4,5 &6 were treated with 100, 200, 400 mg/kgbwt of the extracts respectively. Group 2 serves as toxic control, while group 3 was treated with 25mg/kgbwt silymarin as standard control. Liver marker enzymes ALT, AST, total protein and albumin were assessed and results obtained was extrapolated by performing histological analysis of the liver tissues. The study revealed that treatment of rats with CCl4 caused marked weight loss, induced liver damage through elevated marker enzymes as well as significant decrease in catalase activity. However, administration of graded doses of X. nilotica extract effectively ameliorated the deviation caused by oxidative stress induced liver damage. Pathological examination of the liver tissues also supported the biochemical findings. It was concluded that supplementation of X. nilotica extract was beneficial in modulating the alteration induced in liver and serum variables of rats under the effect of CCl4 induced oxidative stress.

Carbon tetrachloride (CCl4) is a hepatotoxin that causes toxicity in animals on its exposure. Hepatotoxins lead to hepatic demage that is treated by generating antioxidant effect. Synthetic and as well as natural drugs are available for this purpose but phyto-based herbal medicines got paramount importance against drug induced hepatotoxicity Current research was conducted to inspect the protective effect of aqueous extract of Helianthus annuus seeds pre-treatment on the carbon tetrachloride-induced hepatotoxicity in Balb C mice. Study covered the valuation of the enzymatic activity such as alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), alkaline phosphatase (ALP) and Lactate dehydrogenase (LDH) and other biochemical components like bilirubin, total protein, catalase, glutathione (GSH), malondialdehyde bis-(dimethyl acetal) tetra ammonium (MDA). The induction of carbon tetrachloride (CCl4) caused rise in plasma ALAT, ASAT, ALP and LDH. Helianthus annuus seeds extract (HA extract) pre-treatment obliterated CCl4-induced deviations in the activities of these enzymes significantly. Bilirubin level increased whereas total protein contents decreased after the induction of CCl4 this effect was reversed by HA extract. Induction of CCl4 caused increased in MDA level while decrease in GSH and catalase. Helianthus annuus seeds extract also abolished these changes. Helianthus annuus seeds extract pre-treatment also prevented CCl4-induced changes in bilirubin and total protein contents. Carbon tetrachloride treatment resulted in huge hepatic damage. This was prevented by Helianthus annuus seeds extract. These results show that Helianthus annuus seeds extract pre-treatment prevented the mice from CCl4-induced hepatic damage, which visibly shows its defensive effects against hepatic damage.

The aim of the present work is to evaluate the hepatoprotective and antioxidant effect of Nyctanthes arbor –tristis leaf fractions. The petroleum ether, ethylacetate and butanolic fractions of Nyctanthes arbor –tristis leaves were studied to evaluate the hepatoprotective and antioxidant activities in CCl4-induced hepatotoxicity in rats. Oral administration of the fractions at doses of 200 and 400 mg/kg once daily for 10 days significantly restored normalization of serum enzyme levels, viz. glutamate oxaloacetate transaminase (SGOT), glutamate pyruvate transaminase (SGPT) and markers viz. total bilirubin and direct bilirubin and the results were comparable to the effects of Liv 52. The ethylacetate and butanolic extract at the dose of 400 mg/kg was found to be more potent when compared to petroleum ether extract at similar dose. The hepatoprotection is also supported by histopathology of treated animals. In regard to antioxidant activity, ethylacetate and butanolic fractions exhibited a significant effect showing increased levels of enzymatic and non-enzymatic parameters, viz. catalase, GSH, SOD and decreased level of malondialdhyde (MDA). The results of this study strongly indicate that Nyctanthes arbor –tristis leaves have potent antioxidant and hepatoprotective action against CCl4-induced hepatic damage in rats which may be due to the presence phytoconstituents such as flavonoids.

A common challenge that regularly results from oxidative stress is hepatic damage. This condition is characterised by a gradual progression from steatosis to chronic hepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. The study proposed to assess the antioxidant activity and efficacy of Hoya coronaria aqueous extract in preventing CCl4-induced hepatic damage in rats. The DPPH technique was used in the study to assess the extract's antioxidant properties. The rats received dosages of 125 and 250 mg/kg body weight of H. coronaria extract for 14 days, followed by CCl4 exposure. After two weeks, the rats were euthanised for analysis. The results indicated that the extract showed significant antioxidant potential and decreased the impact of CCl4 on hepatic damage markers such as serum aspartate transaminase and alanine transaminase. Moreover, it increased hepatic reduced glutathione and various antioxidant enzymes while reducing malondialdehyde formation induced by CCl4. Additionally, the histopathological analysis demonstrated that H. coronaria extract protected the liver against fatty degeneration and necrosis induced by CCl4 toxicity. These outcomes suggest that H. coronaria extract could be used to prevent ROS-related hepatic damage.

The liver, a vital organ in the body, synthesizes hormones, decomposes RBC in the blood, and regulates glycogen storage. The liver also performs detoxification and is the primary site for drug metabolism. Therefore, hepatoprotective medications are necessary to keep the liver in good health. Ayurvedic drugs are widely used as hepatoprotective agents. The study aims to review the Yakrit (liver) concept in Ayurvedic literature, identify hepatoprotective plants from classical Ayurveda texts, and understand their mode of action on the liver based on existing scientific evidence. The classical Ayurveda literature and electronic databases such as Google Scholar, PubMed, Web of Science, and ARP were searched until October 2022 to find relevant studies using MESH terms. The search of classical Ayurveda texts showed several herbal sources that are advocated for use in liver disorders. The in vivo studies for these plants showed that seven protected the liver from CCl4-induced hepatic damage. Furthermore, the levels of liver enzymes were reduced, and there was evidence of reversal of fatty changes in the liver in mice fed with a high-fat diet. One of the essential organs that aid in sustaining bodily health is the liver. Some plants have been endowed by nature with the ability to treat, prevent, and cure liver disorders while causing fewer negative effects. Herb-based Ayurveda drugs can be a potent source of hepatoprotective agents, though detailed studies need to be conducted to generate more substantial evidence.

Date palm fruit (Phoenix dactylifera: Arecaceae) is rich in essential nutrients and possesses several pharmacological and medicinal activities. The current study aimed to optimize a water bath-assisted extraction method for two cultivars of date palm fruits, Anbara (An) and Reziz (Rz), and investigated the protective effect of the optimized date palm fruit extract against CCl4-induced liver toxicity in relation to oxidative stress, inflammation, apoptosis, and DNA integrity. The optimization process of two date palm fruit cultivars was applied, using response surface methodology through adjusting three “factors”; time, temperature, and rotation, to allow maximum contents of total phenolic (TPC), total flavonoid (TFC), reducing power (FRAP) and scavenging activity (ABTS) of the extract “responses”. Extraction factors’ application significantly enhanced TPC, TFC, FRAP, and ABTS responses by 1.30, 1.23, 3.03, and 2.06-fold, respectively in An and 2.18, 1.71, 1.11, and 2.62-fold, respectively in Rz, in relation to the convectional water extraction. Furthermore, co-administered CCl4 with An or Rz optimized extracts enhanced body weight gain, amended hepatic architecture, and diminished collagen fiber accumulation. Furthermore, An or Rz extracts reduced liver enzymes, hydroxyproline, alpha-fetoprotein (AFP), MDA, inflammatory cytokine (TNF-α, NF-κB) levels, and DNA fragmentation, while increasing deteriorated adiponectin (ADP) and antioxidant enzyme (GSH, GPX, NO, and IFN-γ) levels, relative to CCl4-administered animals. The protective effects of An or Rz-optimized extracts were also evidenced by suppressing hepatic fibrosis and improving liver function and structure via modulating oxidative stress, inflammation, and apoptosis, in CCl4-induced hepatic damage. Hence, the optimized extraction process for the two date palm fruits resulted in extracts which are rich in phenolic and flavonoid contents and with an elevated antioxidant power. The presence of these rich extracts could help to explain their proven hepatoprotective activity against CCl4-induced liver toxicity.

The processing method generally affects the toxicity and biological activity of aged sorghum vinegar. This study investigates the changes in the intermediate Maillard reaction products of sorghum vinegar during ageing and the in vivo hepatoprotective effects of pure melanoidin obtained from it. High-performance liquid chromatography (HPLC) and fluorescence spectrophotometry were utilized to quantify intermediate Maillard reaction products. The CCl4-induced liver damage in rats was used to evaluate the protective role of pure melanoidin in rat liver. Compared with the initial concentration, the 18-month ageing process caused a 1.2- to 3.3-fold increase in the concentrations of intermediate Maillard reaction products, i.e. 5-hydroxymethylfurfural (HMF), 5-methylfurfural (MF), methyglyoxal (MGO), glyoxal (GO) and advanced glycation end products (AGEs). The concentrations of HMF in the aged sorghum vinegar were 6.1-fold higher than the 450 μM limit standard for honey, implying the need for shortening the ageing of the vinegar in practice for safety concerns. Pure melanoidin (Mr>3.5 kDa) demonstrated significant protective effects against CCl4-induced rat liver damage, as evidenced by normalized serum biochemical parameters (transaminases and total bilirubin), suppressing hepatic lipid peroxidation and reactive oxygen species, as well as increasing glutathione amount and restoring antioxidant enzyme activities. Histopathological analysis revealed that melanoidin in vinegar reduced cell infiltration and vacuolar hepatocyte necrosis in rat liver. The findings demonstrated that a shortened ageing process should be considered in practice to ensure the safety of aged sorghum vinegar. Vinegar melanoidin is a potential alternative for the prevention of hepatic oxidative damage. This study demonstrates that the manufacturing process had a profound influence on the generation of vinegar intermediate Maillard reaction products. In particular, it revealed the in vivo hepatoprotective effect of pure melanoidin from aged sorghum vinegar, and provides insight into the in vivo biological activity of melanoidin.

Protective effect of Ceiba pentandra (L) Gaertn on CCl4-induced oxidative stress and liver damage in rats

Mukta Bhasma (MB) is a traditional Ayurvedic preparation for upper respiratory and lower respiratory conditions, eye disorders, powerful cardiac tonic, immune-regulator, mood elevator and known to promote strength, intellect and semen production. Mukta bhasma is prepared using direct heat as a media of transformation. Mukta Bhasma was evaluated for its hepatoprotective and antioxidant activities against Carbon tetrachloride (CCl4) induced liver damage in wistar albino rats. Mukta Bhasma (100mg/kg, 200mg/kg and 300mg/kg) was administered to experimental rats for 10 days. Silymarin (25mg/kg) was given as the standard drug. The hepatoprotective activity was assessed using various serum biochemical parameters like (SGOT), (SGPT), Total and Direct bilirubin (TB and DB), Alkaline phosphatase (ALP), Total Triglycerides (TG) and Total Cholesterol (TC). Lipid peroxidation (LPO), Reduced Glutathione (GSH) and Catalase (CAT) were determined to explain the antioxidant activity of Mukta Bhasma. The substantially elevated levels of SGOT, SGPT, TB and DB, ALP, TG and TC due to CCl4 treatment were restored towards near normal by Mukta bhasma (MB) of 200mg/kg dose. Administration of MB 200mg has shown significant reduction in the biochemical parameters like SGOT, SGPT, ALP, Bilirubin, Total cholesterol and Total triglycerides with significance of p<0.001 in all the parameters when compared to CCl4 group. Also, they were near to the standard Silymarin group. Mukta bhasma 200mg has shown significant (p<0.001) reduction in the LPO level, increase in CAT and GSH representing significant antioxidant activity of Mukta bhasma. The histopathological study showed reduction in fatty degeneration of liver in MB 200mg/kg body weight. Thus, results revealed that Mukta bhasma afford significant hepatoprotective and antioxidant effects in CCl4 induced hepatic damage.

Background: Raphanus sativus is a cruciferous plant which possess free radical scavenging properties that defend the body against the deleterious effects of reactive oxygen species. To evaluate the protective role of Raphanus sativus in CCl4-induced hepatotoxicity through assessment of liver function and oxidative stress markers in albino Wistar rats.
 Material and Methods: Quasi-experimental study was conducted from September 2019 to March 2020 at the Postgraduate Research Laboratory, Isra University, Hyderabad. Non-random purposive sampling was used for selecting Albino Wistar rats. All rats were then divided equally into three different groups: Group A (control group), Group B (CCl4 experimental group), and Group C (CCl4 plus Raphanus sativus group). Blood samples were collected through cardiac puncture followed by hepatic histopathological analysis using light microscope. Data was analyzed using SPSS version 24, with ANOVA and Post hoc Tukey analysis used for comparison of different study variables.
 Results: Statistically significant rise relative liver weight of group B rats compared with the group A and C rats (P <.05). Moreover, statistically significant (P <0.05) rise in serum markers of hepatic functions was observed after CCl4 administration in Group B. Treatment with Raphanus Sativus administration significantly reduced serum levels of LFTs (p<0.05). There was a significant decline in the plasma levels of oxidative markers in group B while marked histopathological changes like necrosis, sinusoidal dilatation and congestion observed among animals of group B.
 Conclusion: Raphanus Sativus exerts an anti-oxidative, anti- and hepatoprotective effect against CCl4-induced hepatic tissue damage.

Exhausted olive pomace (EOP) is produced in olive-pomace oil extractors as a by-product. However, the obtention of bioactive compounds from EOP can reinsert it into the economy as a new bioresource before applying other exploitation ways. The objective of the present study was to investigate the phytochemical differences between aqueous and aqueous acetonic extracts from EOP (AE-EOP and AAE-EOP, respectively) obtained by hydrothermal and ultrasound-assisted extraction, respectively. The in vitro antioxidant activities and the in vivo hepatopreventive potential were evaluated. Using RP-HPLC-ESI-QTOF-MS, the chemical profile revealed that AE-EOP and AAE-EOP showed similar qualitative profiles, with some quantitative differences. Hydroxytyrosol and mannitol were the major compounds of the extracts. The investigation of antioxidant properties in vitro highlighted that AE-EOP was slightly more efficient in scavenging DPPH, ABTS, superoxide, and hydrogen peroxide radicals, when compared to AAE-EOP. Additionally, AE-EOP and AAE-EOP showed dose-dependent suppressive effects on pancreatic lipase activity. In vivo studies showed that AE-EOP and AAE-EOP presented interesting hepatopreventive capacities against CCl4 induced liver injury, as evidenced by (i) the preventive effects against DNA damage, (ii) the normalised hepatic biomarker parameters (ALT, AST, GGT, and LDH) and (iii) the normalised lipid profile (LDL-C, TC, TG, and HDL-C) through diminishing their levels, (iv) which was supported by Oil Red O analysis. Furthermore, AE-EOP and AAE-EOP reduced the oxidative stress in liver tissue by inhibiting lipid peroxidation together with the enhancement of the hepatic antioxidant activities (CAT, SOD and GPx) and GSH content. Additionally, AE-EOP and AAE-EOP exhibited an antifibrotic effect, which was clearly demonstrated by the histopathological examination using Picrosirius red staining. The obtained results support the use of EOP extracts in industries without further purification as antioxidants and against free radical induced damage.

Investigation on medicinal plants' therapeutic potential has gained substantial importance in the discovery of novel effective and safe therapeutic agents. The present study is aimed at investigating the hepatoprotective potential of Seriphidium kurramense methanolic extract (SKM) against carbon tetrachloride- (CCl4-) induced hepatotoxicity in rats. S. kurramense is one of the most imperative plants for its various pharmacological activities. Therefore, this study was aimed at evaluating the hepatoprotective potential against CCl4-induced liver toxicity. The serum samples were analyzed for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) together with the oxidative stress mediator levels as nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), reduced glutathione (GSH), and superoxide dismutase (SOD) as well as peroxidation and H2O2 activity. CCl4 administration resulted in an elevated free radical generation, altered liver marker (AST and ALT) enzymes, reduced antioxidant enzyme, and increased DNA damage. Methanolic extract of S. kurramense decreased CCl4-induced hepatotoxicity by increasing the antioxidant status and reducing H2O2 and nitrate content generation as well as reducing DNA damage. Additionally, SKM reversed the morphological alterations induced by CCl4 in the SKM-treated groups. These results demonstrated that SKM displayed hepatoprotective activity against CCl4-induced hepatic damage in experimental rats.

Background: The main obstacles of silymarin (SIL) application in liver diseases are its low bioavailability, elevated metabolism, rapid excretion in bile and urine, and inefficient intestinal resorption. The study aimed to synthesize and characterize silymarin-conjugated gold nanoparticles (SGNPs) formulation to improve SIL bioavailability and release for potentiating its antifibrotic action. Methods: Both SGNPs and gold nanoparticles (GNPs) were prepared and characterized using standard characterization techniques. The improved formulation was assessed for in vitro drug release study and in vivo study on rats using CCl4 induced hepatic fibrosis model. SIL, SGNPs, and GNPs were administered by oral gavage daily for 30 days. At the end of the study, rats underwent anesthesia and were sacrificed, serum samples were collected for biochemical analysis. Liver tissues were collected to measure the genes and microRNAs (miRNAs) expressions. Also, histopathological and immunohistochemistry (IHC) examinations of hepatic tissues supported these results. Results: The successful formation and conjugation of SGNPs were confirmed by measurements methods. The synthesized nanohybrid SGNPs showed significant antifibrotic therapeutic action against CCl4-induced hepatic damage in rats, and preserved normal body weight, liver weight, liver index values, retained normal hepatic functions, lowered inflammatory markers, declined lipid peroxidation, and activated the antioxidant pathway nuclear factor erythroid-2-related factor 2 (NRF2). The antifibrotic activities of SGNPs mediated through enhancing the hepatic expression of the protective miRNAs; miR-22, miR-29c, and miR-219a which results in suppressed expression of the main fibrosis mediators; TGFβR1, COL3A1, and TGFβR2, respectively. The histopathology and IHC analysis confirmed the anti-fibrotic effects of SGNPs. Conclusions: The successful synthesis of SGNPs with sizes ranging from 16 up to 20 nm and entrapment efficiency and loading capacity 96% and 38.69%, respectively. In vivo studies revealed that the obtained nano-formulation of SIL boosted its anti-fibrotic effects.

PurposeThe polysaccharide of Anoectochilus roxburghii (wall.) Lindl. (ARPS) is one of its important active ingredients. Hepatoprotective effects of ARPS on rat liver injury induced by CCl4 were studied.MethodsARPS was extracted using the ultrasonic method and successfully purified by high-speed counter-current chromatography (HSCCC) with a two-phase aqueous system composed of 12.5% PEG 1000–20% K2HPO4:KH2PO4 (1:1). The HSCCC conditions were optimized, and the structure of ARPS was characterized. The hepatoprotective effects of ARPS against CCl4-induced chronic hepatic injury in SD rats were evaluated.ResultsThe results showed that ARPS was a water-soluble polysaccharide with a molecular weight of 28,518 Da. It was composed of mannose, ribose, glucose, and arabian sugar; its monosaccharide molar ratio was glucose:ribose:arabinose:mannose = 54.24:13.20:1.09:1.00. The purity of ARPS was determined by HPLC to be 96.93%. The intervention effects of ARPS on CCl4-induced hepatic damage model in rats showed that ARPS could effectively reduce the activity of alanine amino transferase and aspartate amino transferase, decrease the content of malondialdehyde and nitric oxide synthesis, and increase the content of glutathione. Pathology revealed that liver plate order, liver cell degeneration, and edema were improved; inflammatory cell infiltration was not observed after ARPS intervention.ConclusionARPS had the function of antioxidant for protecting CCl4-induced injured liver, and the mechanisms were related to anti-lipid peroxidation, which could eliminate oxygen-free radicals and protect liver cells from attacks by free radicals.

Liver disease is a dysfunction that affects all or part of the liver and can lead to death. While the use of medicinal and aromatic plants has been a source of bioactive substances with hepatoprotective properties. This study aims to evaluate the hepatoprotective effect of the methanolic extract of Corrigiola telephiifolia roots on hepatic damage induced by carbon tetrachloride (CCl4) in mice. Animals were treated daily with C. telephiifolia methanolic extract (CTME) at doses of 5 and 10 mg/kg b.w for 60 days. In addition, CCl4 was injected (1 mL/kg, i.p.) for its hepatotoxic effects. At the end of the experiment, the blood of all animals was collected to evaluate biochemical parameters and the liver for histopathological analysis. The administration of CTME showed significant hepatoprotective activity by improving the biochemical parameters and the histological appearance of hepatic cells induced by CCl4. Consequently, C. telephiifolia could be used in the prevention and/or treatment of liver intoxication.

Protective Activity of Cubebin from Syringa oblata Against CCl4-Induced Hepatic Damage in Mice