Toxicity In HepG2 Cell Line Research Articles

Exploring the hepatoprotective properties of citronellol: In vitro and in silico studies on ethanol-induced damage in HepG2 cells.

Citronellol (CT) is a monoterpene alcohol present in the essential oil of plants of the genus Cymbopogon and exhibits diverse pharmacological activities. The aim of the current study was to investigate the hepatoprotective potential of CT against ethanol-induced toxicity in HepG2 cell lines. Silymarin (SIL) was used as a standard drug. MTT, crystal violet assay, DAPI, and PI staining were carried out to assess the effect of ethanol and CT on cell viability. RT-PCR determined the molecular mechanisms of hepatoprotective action of CT. CT ameliorated cell viability and restricted ethanol-induced cell death. DAPI and PI staining showed distinct differences in cell number and morphology. Less cell viability was observed in the diseased group obviously from strong PI staining when compared to the CT- and SIL-treated group. Moreover, CT showed downregulation of interleukin (IL-6), transforming growth factor-beta 1 (TGF-β1), collagen type 1 A 1 (COL1A1), matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase-1 (TIMP-1) and glutathione peroxidase-7 (GPX-7) levels. Molecular docking studies supported the biochemical findings. It is concluded that the cytoprotective activity of CT against ethanol-induced toxicity might be explained by its anti-inflammatory, immunomodulatory, and collagen-regulating effects.

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.

Hepatoprotective activity of Ficus semicordata Buch.-Ham. ex Sm. leaves aqueous extract on D-galactosamine induced toxicity in HepG2 cell line

The leaves of Ficus semicordata Buch-Ham.ex Sm., locally in North India is referred to as Bhumi udumbara have been reported for their traditional use to cure jaundice and various liver ailments. Despite its extensive usage in northeastern states of India, there is a lack of scientific substantiation on the safety and pharmacotherapeutic efficacy. The present study was carried out to evaluate the hepatoprotective activity of F. semicordata (FS) leaves aqueous extract on HepG2 cell line to validate its ethnic claim in the management of liver disorders. D-galactosamine induced HepG2 cell toxicity model was used to evaluate its hepatoprotective activity. The cells were treated with different concentrations of aqueous extracts and dexamethasone as a standard. MTT assay was performed to determine the % inhibition of hepatotoxicity. The result indicates that the toxicity induced by D-galactosamine is reduced by FS aqueous extract group which is better when compared with the standard drug. This study revealed the hepatoprotective potential of F. semicordata on HepG2 cell line.

In vitro hepatoprotective efficacy of extract of Hedychium spicatum rhizome in paracetamol induced toxicity in HepG2 cell line

In vitro hepatoprotective efficacy of extract of Hedychium spicatum rhizome in paracetamol induced toxicity in HepG2 cell line

Psidium guajava Leaf Extracts and Their Quercetin Protect HepG2 Cell Lines Against CCL4 Induced Cytotoxicity.

The present study was carried out to evaluate the in vitro cytoprotective effects of Psidium guajava and their isolated quercetin fraction to reduce the CCl4 (carbon tetrachloride) induced toxicity in HepG2 cell lines (Hepatocellular carcinoma G2). Silymarin was used as a standard drug to compare the protective effects of plant extracts in infected cell lines. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay, cell viability assay, leakage parameters [Aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and Lactate dehydrogenase (LDH)], lipid peroxidation and reduced glutathione (GSH) levels were used to find out the protection of human derived HepG2 cells against CCl4-induced damage. The levels of cytotoxicity, viability and GSH were reduced. While the activities of AST, ALT, LDH and lipid peroxidation was increased in CCl4-treated groups. The treatment of P. guajava and their isolated quercetin fractions (100, 200, 300µg/mL) decreased the elevated levels of all these parameters. The results of the present study suggest that the ethanolic extract of P. guajava leaf and their isolated quercetin fractions can able to reduce the CCl4-induced cytotoxicity in HepG2 cell lines.

UPLC-ESI-MS/MS and HPTLC Method for Quantitative Estimation of Cytotoxic Glycosides and Aglycone in Bioactivity Guided Fractions of Solanum nigrum L.

Solanum nigrum L., is traditionally used for the management of the various liver disorders. Investigating the effect of polarity based fractionation of S. nigrum for its hepatoprotective effect on Hep G2 cells in vitro to provide base of its activity by quantifying in steroidal glycosides responsible for hepatoprotective potential. A new UPLC-ESI-MS/MS method following a high performance thin layer chromatography (HPTLC) has been developed and validated for quantification of steroidal glycosides and aglycone (solasonine, solamargine, and solasodine, respectively). The in vitro antioxidant potential, total phenolics, and flavonoid content were also determined in different fractions. The newly developed UPLC-ESI-MS/MS and HPTLC methods were linear (r2 ≥ 0.99), precise, accurate, and showing recovery more than 97%. The n-butanol enriched fraction of S. nigrum berries was found to be the most potent hepatoprotective fraction against all other fractions as it showed significantly (p < 0.01) better in vitro anti-oxidant potential than other fractions. Quantification by both methods revealed that, content of steroidal glycosides and aglycones are more than 20% in n-butanol fraction as compared to other fractions. The screened steroidal glycoside n-butanol enriched fraction underwent bioefficacy studies against D-galactosamine and H2O2 induced toxicity in HepG2 cell line showing significant (p < 0.05) liver protection. However, developed method can be used for the quality control analysis with respect to targeted metabolites and it can be explored for the pharmacokinetic and pharmacodynamic analysis in future.

Bioactive Guided Fractions of Annona reticulata L. bark: Protection against Liver Toxicity and Inflammation through Inhibiting Oxidative Stress and Proinflammatory Cytokines.

Herbal medicine is popularized worldwide due to its ability to cure the diseases with lesser or no side effects. North Eastern part of India comes under one of the world biodiversity hotspots which is very rich in traditional herbal medicine. Annona reticulata L. (Annonaceae) is one such plant used for the treatment of inflammatory diseases, liver ailments and diabetes by traditional healers. The present study was aimed to scientifically validate this folk knowledge and to develop an herbal remedy through evaluating bioactive guided fractions of A. reticulata (AR) bark against hepatotoxicity and inflammation using in vitro and in vivo models. Results of this study demonstrates that among all fractions of AR bark, methanol extract and its water fraction possess strong anti-oxidant ability and showed protection against CCl4 induced toxicity in HepG2 cell lines and rats. Both the fractions also exhibit dose dependent anti-inflammatory activity against carrageenan induced inflammation in rats. Water fraction showed potent response in the entire tests conducted than methanol extract, which states that polar components of the AR bark methanol extract were responsible for these activities. Further, from the experiments conducted to elucidate the mechanism of action, the results revealed that AR bark showed liver protection and anti-inflammatory response through inhibiting the oxidative stress and inflammatory cytokines.

Combined Effect of Hydroethanolic Extracts of Murraya koenigii and Phyllanthus niruri Leaves on Paracetamol and Ethanol-Induced Toxicity in HepG2 Cell Line

The present study is an attempt to determine the combined hepatoprotective potential of hydroethanolic leaf extracts of Murraya koenigii and Phyllanthus niruri against paracetamol (PCM) and ethanolinduced toxicity in human hepatoma HepG2 cell line. Toxicity in cells was induced by treatment with 15 mM PCM and 50 mM ethanol for 24 h as manifested by a significant ( P < 0.05) decrease in cell viability, increase in the leakage of serum glutamate oxaloacetate transaminase and serum glutamate pyruvate in culture medium, increase in lipid peroxidation and reduction in reduced glutathione in cell lysate. These alterations were significantly ameliorated when cells were treated with a combination of hydroethanolic leaf extracts of M. koenigii and P. niruri , and silymarin during both prophylactic and curative studies. Both post-treatment (curative) and pre-treatment (prophylactic) with the combination of plant extracts were able show effective hepatoprotection. This was also evident during morphological studies. The combination of plant extracts thus holds immense potential for future use as a hepatoprotectant.

Combined Effect of Hydroethanolic Extracts of &lt;I&gt;Murraya Koenigii&lt;/I&gt; and &lt;I&gt;Phyllanthus Niruri&lt;/I&gt; Leaves on Paracetamol and Ethanol-induced Toxicity in HepG2 Cell Line

Combined Effect of Hydroethanolic Extracts of &lt;I&gt;Murraya Koenigii&lt;/I&gt; and &lt;I&gt;Phyllanthus Niruri&lt;/I&gt; Leaves on Paracetamol and Ethanol-induced Toxicity in HepG2 Cell Line

Combined Effect of Hydroethanolic Extracts of &lt;I&gt;Murraya Koenigii&lt;/I&gt; and &lt;I&gt;Phyllanthus Niruri&lt;/I&gt; Leaves on Paracetamol and Ethanol-induced Toxicity in HepG2 Cell Line

Combined Effect of Hydroethanolic Extracts of &lt;I&gt;Murraya Koenigii&lt;/I&gt; and &lt;I&gt;Phyllanthus Niruri&lt;/I&gt; Leaves on Paracetamol and Ethanol-induced Toxicity in HepG2 Cell Line

Antioxidant and hepatoprotective activity of Fagonia schweinfurthii (Hadidi) Hadidi extract in carbon tetrachloride induced hepatotoxicity in HepG2 cell line and rats

Ethnopharmacological relevanceThe whole plant of Fagonia schweinfurthii (Hadidi) Hadidi (Family: Zygophyllaceae) is used in variety of diseases including hepatic ailments in deserts and dry areas of India. Aim of the studyTo evaluate antioxidant and hepatoprotective activity of ethanolic extract from Fagonia schweinfurthii (Hadidi) Hadidi (FSEE) in carbon tetrachloride (CCl4) induced hepatotoxicity in HepG2 cell line and rats. Materials and methodsIn vitro antioxidant activity was determined by DPPH, ABTS radicals and hydrogen peroxide methods. In vitro cytotoxicity and hepatoprotective potential of FSEE were evaluated using HepG2 cells. Based on the cytotoxicity assay, FSEE (50, 100, 200µg/ml) was assessed for hepatoprotective potential against CCl4 induced toxicity in HepG2 cell line by monitoring cell viability, aspartate aminotransferase (AST), alanine aminotransaminase (ALT), lactate dehydrogenase (LDH) leakage, lipid peroxidation (LPO) and glutathione level (GSH). Further, in vivo hepatoprotective activity of FSEE was evaluated against CCl4 induced hepatotoxicity in male Wistar albino rats. Rats were pre-treated with FSEE (200mg, 400mgkg−1day−1 p.o.) for 7 days followed by a single dose of CCl4 (1.0ml/kg, i.p.) on 8th day. Silymarin was used as positive control. After 24h of CCl4 administration, various biochemical parameters like aspartate aminotransferase (AST), alanine aminotransaminase (ALT), alkaline phosphatase (ALP), total bilirubin (TB) and total protein (TP) levels were estimated in serum. The antioxidant parameters like superoxide dismutase (SOD) activity, catalase (CAT) activity, glutathione (GSH) content and malondialdehyde (MDA) level in the liver homogenate were determined. Histopathological changes in the liver of different groups were also studied. ResultsThe FSEE possessed strong antioxidant activity in vitro. The results indicated that CCl4 treatment caused a significant decrease in cell viability. The CCl4-induced changes in the HepG2 cells were significantly ameliorated by treatment of the FSEE. FSEE significantly prevented CCl4 induced elevation of AST, ALT, ALP, TB, and CCl4 induced decrease in total protein in rats. FSEE treated rat liver anti-oxidant parameters (SOD, CAT, MDA and GSH,) were significantly antagonized for the pro-oxidant effect of CCl4. Histopathological studies also supported the protective effect of FSEE. ConclusionThe results of this study revealed that FSEE has significant hepatoprotective activity. This effect may be due to the ability of the extract to inhibit lipid peroxidation and increase in the anti-oxidant enzymatic activity.

In vitro hepatoprotective activity of Corchorus depressus L. against CCl4 induced toxicity in HepG2 cell line

AimTo evaluate in vitro hepatoprotective activity of ethanolic extract from Corchorus depressus (CDEE) against CCl4 induced toxicity in HepG2 cell line. MethodsIn vitro cytotoxicity and hepatoprotective potential of CDEE were evaluated using HepG2 cells. Based on the cytotoxicity assay, CDEE (50, 100, 200 μg/ml) was assessed for hepatoprotective potential against CCl4 induced toxicity by monitoring cell viability, aspartate aminotransferase (AST), alanine aminotransaminase (ALT), lactate dehydrogenase (LDH) leakage, lipid peroxidation (LPO) and glutathione level (GSH). ResultsThe results indicated that CCl4 treatment caused a significant decrease in cell viability. In addition, the toxin treatment initiated lipid peroxidation (LPO), caused leakage of enzymes like transaminase (AST & ALT) and LDH with a significant decrease in GSH levels in HepG2 cells. It was observed that CDEE effectively alleviated the changes induced by CCl4 in a concentration-dependent manner. ConclusionOur study revealed that CDEE has potent cytoprotective effect against CCl4 induced toxicity in HepG2 cell line and which may be attributed to decrease in CCl4 induced reactive oxygen species levels and resultant oxidative stress.

Mangiferin, a dietary xanthone protects against mercury-induced toxicity in HepG2 cells.

Mercury is one of the noxious heavy metal environmental toxicants and is a cause of concern for human exposure. Mangiferin (MGN), a glucosylxanthone found in Mangifera indica, reported to have a wide range of pharmacological properties. The objective of this study was to evaluate the cytoprotective potential of MGN, against mercury chloride (HgCl(2) ) induced toxicity in HepG2 cell line. The cytoprotective effect of MGN on HgCl(2) induced toxicity was assessed by colony formation assay, while antiapoptotic effect by fluorescence microscopy, flow cytometric DNA analysis, and DNA fragmentation pattern assays. Further, the cytoprotective effect of MGN against HgCl(2) toxicity was assessed by using biochemical parameters like reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) by spectrophotometrically, mitochondrial membrane potential by flowcytometry and the changes in reactive oxygen species levels by DCFH-DA spectrofluoremetric analysis. A significant increase in the surviving fraction was observed with 50 μM of MGN administered two hours prior to various concentrations of HgCl(2) . Further, pretreatment of MGN significantly decreased the percentage of HgCl(2) induced apoptotic cells. Similarly, the levels of ROS generated by the HgCl(2) treatment were inhibited significantly (P < 0.01) by MGN. MGN also significantly (P < 0.01) inhibited the HgCl(2) induced decrease in GSH, GST, SOD, and CAT levels at all the post incubation intervals. Our study demonstrated the cytoprotective potential of MGN, which may be attributed to quenching of the ROS generated in the cells due to oxidative stress induced by HgCl(2) , restoration of mitochondrial membrane potential and normalization of cellular antioxidant levels.

Isolation, characterization and antioxidative effect of phyllanthin against CCl 4-induced toxicity in HepG2 cell line

The present study was an attempt to investigate the hepatoprotective and antioxidative property of Phyllanthus amarus ( P. amarus) extract and phyllanthin. Phyllanthin, one of the active lignin present in this plant species was isolated from the aerial parts, by silica gel column chromatography employing gradient elution with hexane–ethyl acetate solvent mixture. It was obtained in high yields (1.23%), compared to reported procedures and the purity was ascertained by HPTLC and reversed-phase HPLC analysis. Characterization of phyllanthin was done by mp, UV–Visible spectrophotometry, elemental analysis, FT-IR, 1H NMR, 13C NMR and mass spectral analysis. Free radical scavenging activity of P. amarus extract and phyllanthin was also examined using DPPH assay. The protective effect of P. amarus extract and phyllanthin was studied on CCl 4-induced toxicity in human hepatoma HepG2 cell line. The results indicated that CCl 4 treatment caused a significant decrease in cell viability. In addition, the toxin treatment initiated lipid peroxidation (LPO), caused leakage of enzymes like alanine transaminase (ALT) and lactate dehydrogenase (LDH) with a significant decrease in glutathione (GSH) levels. It was observed that phyllanthin effectively alleviated the changes induced by CCl 4 in a concentration-dependent manner, with much smaller strengths as compared to P. amarus extract.

Cytoprotective and antigenotoxic potential of Mangiferin, a glucosylxanthone against cadmium chloride induced toxicity in HepG2 cells

Mangiferin (MGN), a glucosylxanthone present in large amounts in the leaves and edible mango fruits of Mangifera indica. Here, we report about MGN’s potential for mitigating cadmium chloride (CdCl 2) induced cytotoxic and genotoxic effects in HepG2 cells growing in vitro. The cytoprotective potential was assessed by MTT, clonogenic and apoptotic assays, while antigenotoxic effect by micronucleus and comet assay. The established cytotoxic and genotoxic effects were well indicated after CdCl 2 treatment and was mitigated by pretreatment with MGN. MGN prior to CdCl 2 treatment increased the cell survival (MTT), surviving fraction (clonogenic assay) and inhibited sub-G 1 population (flow cytometric analysis). Further, inhibition of CdCl 2 induced apoptotic cell death by MGN was confirmed by microscopic and DNA fragmentation assays. A significant (p < 0.01) reduction in the micronuclei frequency and comet parameters after MGN pretreatment to CdCl 2 clearly indicated the antigenotoxic potential. Similarly, the reactive oxygen species generated by the CdCl 2 treatment were inhibited significantly (p < 0.001) by MGN. Taken together, our study revealed that MGN has potent cytoprotective and antigenotoxic effect against CdCl 2 induced toxicity in HepG2 cell line and which may be attributed to decrease in CdCl 2 induced reactive oxygen species levels and resultant oxidative stress.