scholarly journals Effects of Pyrene on Human Liver HepG2 Cells: Cytotoxicity, Oxidative Stress, and Transcriptomic Changes in Xenobiotic Metabolizing Enzymes and Inflammatory Markers with Protection Trial Using Lycopene

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Jin-Kui Ma ◽  
Walaa Fathy Saad Eldin ◽  
Waleed Rizk El-Ghareeb ◽  
Abdelazim Elsayed Elhelaly ◽  
Mariam H. E. Khedr ◽  
...  
Keyword(s):  
Antioxidant Enzymes ◽  
Mrna Expression ◽  
Hepg2 Cells ◽  
Inflammatory Markers ◽  
Human Liver ◽  
Regulatory Elements ◽  
Related Factor ◽  
Dependent Manner ◽  
Metabolizing Enzymes ◽  
Xenobiotic Metabolizing Enzymes

Pyrene is one of the major polycyclic aromatic hydrocarbons formed during heat treatment of meat and in car exhausts; however, few studies have investigated pyrene-induced adverse effects on human cell lines. This study aimed at the investigation of pyrene-induced cytotoxicity and oxidative damage in human liver HepG2 cells at environmentally relevant concentrations. Pyrene-induced changes in mRNA expression of xenobiotic metabolizing enzymes (XMEs), xenobiotic transporters, antioxidant enzymes, and inflammatory markers were investigated using real-time PCR. As a protection trial, the ameliorative effects of lycopene, a carotenoid abundantly found in tomato, were investigated. The possible mechanisms behind such effects were examined via studying the co exposure effects of pyrene and lycopene on regulatory elements including the aryl hydrocarbon receptor (Air) and elytroid 2-related factor 2 (RF). The achieved results indicated that pyrene caused significant cytotoxicity at 50 n, with a clear production of reactive oxygen species (ROS) in a dose-dependent manner. Pyrene upregulated mRNA expression of phase I enzymes including CYP1A1, 1A2, and CYP1B1 and inflammatory markers including TNFα and Cox2. However, pyrene significantly downregulated phase II enzymes, xenobiotic transporters, and antioxidant enzymes. Interestingly, lycopene significantly reduced pyrene-induced cytotoxicity and ROS production. Moreover, lycopene upregulated detoxification and antioxidant enzymes, probably via its regulatory effects on Air- and RF-dependent pathways.

scholarly journals Modulatory effects of rutin on the expression of cytochrome P450s and antioxidant enzymes in human hepatoma cells

2016 ◽  
Vol 66 (4) ◽  
pp. 491-502 ◽  
Author(s):  
Sedar Karakurt
Keyword(s):  
Antioxidant Enzymes ◽  
Phase Ii ◽  
Hepg2 Cells ◽  
Cytochrome P450s ◽  
Dependent Manner ◽  
Protective Effects ◽  
Human Hepatoma Cells ◽  
Metabolizing Enzymes ◽  
Protein Expressions ◽  
Dose Dependent

Abstract Expression of a drug and xenobiotic metabolizing enzymes, cytochrome P450s (CYPs), and antioxidant enzymes can be modulated by various factors. The flavonoid rutin was investigated for its anti-carcinogen and protective effects as well as modulatory action on CYPs and phase II enzymes in human hepatocellular carcinoma cells. Rutin inhibited proliferation of HEPG2 cells in a dose-dependent manner with the IC50 value of 52.7 μmol L-1 and invasion of HEPG2 cells (21.6 %, p = 0.0018) and colony formation of those invaded cells (57.4 %, p < 0.0001). Rutin treatment also significantly increased early/late-stage apoptosis in HEPG2 cells (28.9 %, p < 0.001). Treatment by rutin significantly inhibited protein expressions of cytochrome P450-dependent CYP3A4 (75.3 %, p < 0.0001), elevated CYP1A1 enzymes (1.7-fold, p = 0.0084) and increased protein expressions of antioxidant and phase II reaction catalyzing enzymes, NQO1 (2.42-fold, p < 0.0001) and GSTP1 (2.03-fold, p < 0.0001). Besides, rutin treatment significantly inhibited mRNA expression of CYP3A4 (73.2 %, p=0.0014). Also, CYP1A1, NQO1 and GSTP1 mRNA expressions were significantly increased 2.77-fold (p = 0.029), 4.85- fold (p = 0.0051) and 9.84-fold (p < 0.0001), respectively.

Liraglutide Ameliorates Lipotoxicity-Induced Oxidative Stress by Activating the NRF2 Pathway in HepG2 Cells

2020 ◽  
Vol 52 (07) ◽  
pp. 532-539 ◽  
Author(s):  
Chong-gui Zhu ◽  
Ying Luo ◽  
Hao Wang ◽  
Jun-Yi Li ◽  
Jie Yang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Enzymes ◽  
Mrna Expression ◽  
Hepg2 Cells ◽  
Target Genes ◽  
Nuclear Translocation ◽  
Thiobarbituric Acid ◽  
Related Factor ◽  
Protective Effects ◽  
Alcoholic Fatty Liver

AbstractAlthough glucagon-like peptide-1 (GLP-1) analogue has been reported to suppress oxidative stress in non-alcoholic fatty liver disease (NAFLD), an effective therapeutic agent for NAFLD is currently unavailable. Therefore, in this study, we aimed to investigate the protective effects of the GLP-1 analogue liraglutide against lipotoxicity-induced oxidative stress in HepG2 cells and to elucidate the underlying mechanisms. HepG2 cells were cultured for 48 hours and treated with a free fatty acid (FFA) mixture: FFA mixture and liraglutide or FFA mixture, liraglutide, and exendin (9–39). Lipid accumulation was examined by oil red O staining. Oxidative stress was assessed by measuring the levels of intracellular reactive oxygen species using 2′,7′-dichlorofluorescein diacetate and thiobarbituric acid-reactive substances, whereas antioxidant capacity was assessed by measuring the activity of superoxide dismutase and catalase. Expression of the nuclear factor erythroid-2-related factor 2 (NRF2) gene and the genes encoding antioxidant enzymes was analyzed using quantitative RT-PCR. Cellular and nuclear NRF2 expression levels were assessed using immunofluorescence cell staining and western blotting. Liraglutide treatment reduced high fat-induced lipid formation and the levels of oxidative stress markers and increased antioxidant enzyme activity in HepG2 cells. Liraglutide treatment increased the mRNA expression of NRF2 target genes, induced NRF2 nuclear translocation, and increased nuclear NRF2 levels without altering NRF2 mRNA expression. Collectively, these results indicate that liraglutide exhibits a protective effect against lipotoxicity-induced oxidative stress, possibly via modulation of NRF2 and expression of antioxidant enzymes in liver cells.

scholarly journals S-Allyl Cysteine Alleviates Hydrogen Peroxide Induced Oxidative Injury and Apoptosis through Upregulation of Akt/Nrf-2/HO-1 Signaling Pathway in HepG2 Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-14 ◽  
Author(s):  
Chitra Basu ◽  
Runa Sur
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Oxidative Damage ◽  
Hepg2 Cells ◽  
Liver Diseases ◽  
Heme Oxygenase 1 ◽  
Related Factor ◽  
Dependent Manner ◽  
Cell Viability Assay

Hydrogen peroxide (H2O2) mediated oxidative stress leading to hepatocyte apoptosis plays a pivotal role in the pathophysiology of several chronic liver diseases. This study demonstrates that S-allyl cysteine (SAC) renders cytoprotective effects on H2O2 induced oxidative damage and apoptosis in HepG2 cells. Cell viability assay showed that SAC protected HepG2 cells from H2O2 induced cytotoxicity. Further, SAC treatment dose dependently inhibited H2O2 induced apoptosis via decreasing the Bax/Bcl-2 ratio, restoring mitochondrial membrane potential (∆Ψm), inhibiting mitochondrial cytochrome c release, and inhibiting proteolytic cleavage of caspase-3. SAC protected cells from H2O2 induced oxidative damage by inhibiting reactive oxygen species accumulation and lipid peroxidation. The mechanism underlying the antiapoptotic and antioxidative role of SAC is the induction of the heme oxygenase-1 (HO-1) gene in an NF-E2-related factor-2 (Nrf-2) and Akt dependent manner. Specifically SAC was found to induce the phosphorylation of Akt and enhance the nuclear localization of Nrf-2 in cells. Our results were further confirmed by specific HO-1 gene knockdown studies which clearly demonstrated that HO-1 induction indeed played a key role in SAC mediated inhibition of apoptosis and ROS production in HepG2 cells, thus suggesting a hepatoprotective role of SAC in combating oxidative stress mediated liver diseases.

Zederone, a Sesquiterpene from Curcuma elata Roxb, is Hepatotoxic in Mice

2013 ◽  
Vol 32 (6) ◽  
pp. 454-462 ◽  
Author(s):  
Prapapan Pimkaew ◽  
Kanoknetr Suksen ◽  
Koravit Somkid ◽  
Ratchanaporn Chokchaisiri ◽  
Surawat Jariyawat ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Messenger Rna ◽  
Lethal Dose ◽  
Related Factor ◽  
Metabolizing Enzymes ◽  
Centrilobular Necrosis ◽  
Cytokine Tumor Necrosis Factor ◽  
Antioxidant Gene Expression ◽  
Liver Enlargement ◽  
Factor Α

The present study aimed to investigate the hepatotoxicity of zederone isolated from Curcuma elata in mice. Adult male mice were intraperitoneally injected with a single dose of zederone (50-300 mg/kg body weight [BW]). Twenty-four hours after the injection, zederone induced liver enlargement with scattered white foci over the organ. The medium lethal dose (LD50) value at 24 hours of zederone was approximately 223 mg/kg BW. Hepatic centrilobular necrosis with marked increases in plasma alanine transaminase activity and total bilirubin levels was observed. Zederone at a dose of 200 mg/kg BW markedly decreased the activity of superoxide dismutase and the hepatic glutathione content, whereas the activity of catalase was not altered. The compound at this dose also increased the messenger RNA (mRNA) expression of Cyp2b10 and Cyp3a11, which are the main drug-metabolizing enzymes in the liver. The mRNA expression of proinflammatory cytokine tumor necrosis factor α was increased. The nuclear factor-E2-related factor 2 protein, which is the transcription factor regulating the antioxidant gene expression, was decreased. The histopathology of massive hepatic centrilobular necrosis with an increase in the expression of cytochrome P450 (Cyp) suggests that the possible potentiation of zederone-induced hepatotoxicity implicated the induction of Cyps, which leads to the formation of biological reactive metabolites and that cause the oxidative stress and liver cell injuries.

Abstract 067: Downregulation Of Vascular Factor-erythroid 2-related Factor-2 And Associated Antioxidant Enzymes In Hypertension

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Rheure A Lopes ◽  
Karla B Neves ◽  
Augusto Montezano ◽  
Rita Tostes ◽  
Rhian Touyz
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Enzymes ◽  
Mrna Expression ◽  
Vascular Dysfunction ◽  
Ros Generation ◽  
Related Factor ◽  
Mrna Levels ◽  
Ang Ii ◽  
Peroxiredoxin 1 ◽  
Anti Oxidant

Oxidative stress plays an important role in vascular dysfunction in hypertension. While mechanisms regulating vascular pro-oxidants are emerging, there is a paucity of information on anti-oxidant systems. Factor-erythroid 2-related factor-2 (Nrf2) is a master regulator of antioxidants and its role in hypertension remains elusive. We assessed vascular Nrf2 in hypertension by studying mesenteric vessels and VSMCs from WKY and SHRSP rats. Cells were stimulated with Ang II (10-7M) in the absence/presence of Nrf2 activators (bardoxolone or L-sulforaphane). ROS generation was assessed by chemiluminescence and amplex red. mRNA expression of anti-oxidant enzymes was assessed by qPCR. Nrf2 activity was analyzed by ELISA. Nrf2 activity was decreased in arteries (18%) and VSMCs (48%) in SHRSP (p<0.05 vs WKY). mRNA levels of antioxidant enzymes were reduced in SHRSP (SOD 1 (64%), catalase (60%), peroxiredoxin 1 (75%) and glutathione peroxidase (54%) Ang II increased Nrf2 activity in VSMCs from WKY (197%, 4h) and SHRSP (44%, 4h) (p<0.05, vs. vehicle). This was associated with increased antioxidant mRNA expression in WKY rats (SOD1-32%, catalase-42%, thioredoxin-71%, peroxiredoxin 1-90%, quinone oxidoreductase-84%; p<0.05 vs. vehicle) but not in SHRSP. ROS production and glucose-6-phosphate dehydrogenase (source of NADPH) mRNA levels were increased in SHRSP. Ang II-induced ROS generation in VSMCs from WKY and SHRSP was blocked by Nrf2 activators. Vascular function assessment, by wire myography, demonstrated that increased contractility (Emax Phe: WKY 113.4±5,67 vs. SHRSP 159.0±8.29) and decreased endothelial-dependent relaxation (Emax ACh: WKY 88.7±3.13 vs. SHRSP 74.7±3.25, p<0.05) in SHRSP were corrected by bardoxolone and L-sulforaphane. In conclusion, vascular dysfunction in SHRSP is associated with oxidative stress, decreased Nrf2 activity and reduced Nrf2-regulated antioxidant enzymes. A similar molecular phenotype was observed in Ang II-stimulated VSMCs. Nrf-2 agonists ameliorated vascular dysfunction in SHRSP. Our findings suggest that Nrf-2 downregulation may contribute to redox-sensitive vascular dysfunction and could be a therapeutic target in hypertension. Financial Support: ScWB.

Resveratrol Regulates Nrf2-Mediated Expression of Antioxidant and Xenobiotic Metabolizing Enzymes in Pesticides-Induced Parkinsonism

2020 ◽  
Vol 27 (10) ◽  
pp. 1038-1045
Author(s):  
Mohd Sami Ur Rasheed ◽  
Manish Kumar Tripathi ◽  
Devendra Kumar Patel ◽  
Mahendra Pratap Singh
Keyword(s):  
Nuclear Translocation ◽  
Neuroblastoma Cells ◽  
Related Factor ◽  
Quinone Oxidoreductase ◽  
Metabolizing Enzymes ◽  
Cyp2d6 Activity ◽  
Xenobiotic Metabolizing Enzymes ◽  
Dopamine Content ◽  
Nrf2 Activation ◽  
Induced Changes

Background: Combined maneb (MB) and paraquat (PQ), two widely used pesticides, increases oxidative stress leading to Parkinsonism. Xenobiotic metabolizing enzymes, cytochrome P450 (CYP) 2D6 and its mouse ortholog Cyp2d22 protect against Parkinsonism. Resveratrol, an antioxidant, restores antioxidant defense system through the activation of nuclear factor erythroid 2- related factor 2 (Nrf2). However, a crosstalk between Cyp2d22/CYP2D6-mediated protection and resveratrol-induced Nrf2 activation leading to neuroprotection is not yet elucidated. Objective: The study aimed to decipher the effect of resveratrol on Nrf2 activation and expression of its downstream mediators, nicotinamide adenine dinucleotide phosphate quinone oxidoreductase 1 (NQO1) and thioredoxin 1 (Trx1) along with Cyp2d22/CYP2D6 activity in combined MB and PQ mouse model of Parkinsonism and differentiated neuroblastoma cells. Results: MB and PQ reduced the dopamine content (mouse) and Cyp2d22/CYP2D6 activity (mouse/neuroblastoma cells) and increased the nuclear translocation of Nrf2 and expression of NQO1 and Trx1 (both). Resveratrol ameliorated pesticides-induced changes in dopamine content and Cyp2d22/CYP2D6 activity. It was found to promote nuclear translocation of Nrf2 and expression of NQO1 and Trx1 proteins. Since Cyp2d22/CYP2D6 inhibitor (ketoconazole/quinidine) per se reduced Cyp2d22/CYP2D6 activity and dopamine content, it was found to substantially increase the pesticides-induced reduction in Cyp2d22/CYP2D6 activity and dopamine content. Inhibitors normalized the pesticides induced changes in Nrf2 translocation and NQO1 and Trx1 levels in pesticides treated groups. Conclusion: The results suggest that resveratrol promotes the catalytic activity of xenobiotic metabolizing enzyme, Cyp2d22/CYP2D6, which partially contributes to Nrf2 activation in pesticides- induced Parkinsonism.

scholarly journals Effects of Flutriafol Fungicide on the Lipid Accumulation in Human Liver Cells and Rat Liver

Foods ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1346
Author(s):  
Hyuk-Cheol Kwon ◽  
Hyejin Sohn ◽  
Do-Hyun Kim ◽  
Chang-Hee Jeong ◽  
Dong-Wook Kim ◽  
...  
Keyword(s):  
Lipid Accumulation ◽  
Hepg2 Cells ◽  
Human Liver ◽  
Liver Toxicity ◽  
Fatty Infiltration ◽  
Liver Cells ◽  
Related Factor ◽  
Mrna Levels ◽  
Human Liver Cells ◽  
Sd Rats

Flutriafol (FTF) is a triazole fungicide that can cause liver toxicity through the ingestion of its residues in food and water. However, little is known about the liver toxicity of FTF, particularly nonalcoholic fatty liver disease (NAFLD) in humans. Therefore, the purpose of this study was to investigate whether FTF induces NAFLD in human liver cells and animal liver. HepG2 cells and Sprague Dawley (SD) rats were treated with FTF at doses of 0–640 µM for 24 h and 0–150 mg/kg bw/day for 28 days, respectively. FTF (80, 160, and 320 µM) treatment to cells induced lipid accumulation. FTF (80 and 160 µM)-treated cells had higher levels of cytochrome P450 enzymes and reactive oxygen species and increased mitochondrial membrane potential loss than the control. FTF also increased the mRNA levels of antioxidant enzymes through oxidative stress and nuclear factor erythroid 2-related factor 2 pathways in HepG2 cells. However, a higher level of FTF (320 µM) induced apoptosis. The treatment of SD rats with FTF (2.5–150 mg/kg bw/day) induced fatty infiltration in the liver by impairing liver metabolism and inducing apoptosis. Therefore, our data suggest that human exposure to FTF residues may be a risk factor for liver diseases, such as NAFLD.

scholarly journals Statins Activate Human PPAR Promoter and Increase PPAR mRNA Expression and Activation in HepG2 Cells

PPAR Research ◽  
2008 ◽  
Vol 2008 ◽  
pp. 1-11 ◽  
Author(s):  
Makoto Seo ◽  
Ikuo Inoue ◽  
Masaaki Ikeda ◽  
Takanari Nakano ◽  
Seiichiro Takahashi ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Hepg2 Cells ◽  
Promoter Activity ◽  
Transcriptional Activity ◽  
Statin Treatment ◽  
Expression Vectors ◽  
Peroxisome Proliferator ◽  
Dependent Manner ◽  
Peroxisome Proliferator Activated Receptor ◽  
Protein Levels

Statins increase peroxisome proliferator-activated receptor (PPAR) mRNA expression, but the mechanism of this increased PPAR production remains elusive. To examine the regulation of PPAR production, we examined the effect of 7 statins (atorvastatin, cerivastatin, fluvastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin) on human PPAR promoter activity, mRNA expression, nuclear protein levels, and transcriptional activity. The main results are as follows. (1) Majority of statins enhanced PPAR promoter activity in a dose-dependent manner in HepG2 cells transfected with the human PPAR promoter. This enhancement may be mediated by statin-induced HNF-4. (2) PPAR mRNA expression was increased by statin treatment. (3) The PPAR levels in nuclear fractions were increased by statin treatment. (4) Simvastatin, pravastatin, and cerivastatin markedly enhanced transcriptional activity in 293T cells cotransfected with acyl-coenzyme A oxidase promoter and PPAR/RXR expression vectors. In summary, these data demonstrate that PPAR production and activation are upregulated through the PPAR promoter activity by statin treatment.

scholarly journals AQP9 expression and phosphorylation affect arsenic uptake and tolerance in human liver hepatocellular cells

2021 ◽  
Author(s):  
Rong Chen ◽  
Xiaozhen Wang ◽  
Qianqian Wang ◽  
Yu Hong ◽  
Xianrong Xu ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Mammalian Cells ◽  
Human Liver ◽  
Mapk Pathway ◽  
Specific Inhibitor ◽  
Experimental Period ◽  
Dependent Manner ◽  
Arsenic Uptake ◽  
Arsenic Tolerance ◽  
Arsenic Accumulation

Abstract Background Little is known about the signaling mechanisms involved in arsenic tolerance or detoxification, although the water channel protein aquaporin-9 (AQP9) and components of the mitogen-activated protein kinase (MAPK) pathway have putative roles. Elucidation of the mechanisms of resistance in mammalian cells would be helpful in developing effective, arsenic-based therapeutic strategies. Methods The association between AQP9 and arsenic accumulation and tolerance was investigated in arsenic-sensitive human liver hepatocellular cells (HepG2) and arsenic-resistant HepG2 (AsHepG2) cells. Results The IC50 value for arsenic of AsHepG2 cells (15.59 µM) was significantly higher than that of HepG2 cells (7.33 µM; Ρ < 0.05), and AsHepG2 cells accumulated significantly low levels of arsenic after treatment with sodium arsenite (NaAsO2; Ρ < 0.01). Arsenic accumulation in AsHepG2 cells reached a plateau 6 h after treatment, while in HepG2 cells it continued to increase throughout the experimental period. Additionally, intracellular arsenic accumulation in AQP9-overexpressing AsHepG2 cells significantly increased within 10 h of treatment, whereas in HepG2 cells it increased throughout the experimental period. The level of AQP9 protein in AsHepG2 cells decreased in a concentration-dependent manner, but was not markedly different in HepG2 cells. Furthermore, after NaAsO2 treatment the level of phosphorylated AQP9 and p38 was significantly increased with time in the two cell lines. Partial inhibition of p38 activity by the specific inhibitor SB203580 did not significantly affect AQP9 protein expression or phosphorylation. Conclusion AQP9 expression and its state of phosphorylation are closely related to arsenic uptake and may regulate cellular arsenic tolerance by reducing its uptake rate. p38 may have a limited role in the regulation of AQP9 phosphorylation in AsHepG2 cells.

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