Differential response of antioxidant defense in HepG2 cells on exposure of Livotrit®, in a concentration dependent manner

Background: Arctigenin is a lignan found in Arctium lappa L. (Asteraceae) that displays anti-inflammatory activities. Previous studies showed that the crude extract of A. Lappa has antitumor activity in human liver carcinoma, lung and stomach cancer cells. The aim of this study was to obtain arctigenin from A. lappa L., as well as to evaluate its antiproliferative effects in cells of liver carcinoma (HepG2) and fibroblasts (NIH/3T3). Methods: Arctigenin was obtained from the hydrolysis of arctiin, which was isolated from the crude extract of A. lappa. The effects of arctigenin and arctiin on HepG2 cell viability and cell adhesion were analyzed by MTT method. Adhesion assay was also carried out to evaluate the antitumor activity. Results: Our results showed that the analytical process to obtain arctigenin was fast and easy. In vitro experiments showed that arctigenin (107-269 μM) decreased HepG2 cells viability and did not cause cytotoxicity on NIH/3T3 cells. Arctigenin (27-269 μM) demonstrated anti-adhesion in HepG2 cells in a concentration-dependent manner, when compared with control. Conclusion: These results suggest a promising pharmacological activity for arctigenin as an antiproliferative compound.

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Cholesterol lowering effects of mono-lactose-appended β-cyclodextrin in Niemann–Pick type C disease-like HepG2 cells

Beilstein Journal of Organic Chemistry ◽

10.3762/bjoc.11.224 ◽

2015 ◽

Vol 11 ◽

pp. 2079-2086 ◽

Cited By ~ 12

Author(s):

Keiichi Motoyama ◽

Yumi Hirai ◽

Rena Nishiyama ◽

Yuki Maeda ◽

Taishi Higashi ◽

...

Keyword(s):

Hepg2 Cells ◽

Cholesterol Content ◽

High Dose ◽

Dependent Manner ◽

Lysosomal Storage ◽

Concentration Dependent Manner ◽

Cholesterol Lowering ◽

Type C ◽

Intracellular Cholesterol ◽

Niemann Pick

The Niemann–Pick type C disease (NPC) is one of inherited lysosomal storage disorders, emerges the accumulation of unesterified cholesterol in endolysosomes. Currently, 2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) has been applied for the treatment of NPC. HP-β-CyD improved hepatosplenomegaly in NPC patients, however, a high dose of HP-β-CyD was necessary. Therefore, the decrease in dose by actively targeted-β-CyD to hepatocytes is expected. In the present study, to deliver β-CyD selectively to hepatocytes, we newly fabricated mono-lactose-appended β-CyD (Lac-β-CyD) and evaluated its cholesterol lowering effects in NPC-like HepG2 cells, cholesterol accumulated HepG2 cells induced by treatment with U18666A. Lac-β-CyD (degree of substitution of lactose (DSL) 1) significantly decreased the intracellular cholesterol content in a concentration-dependent manner. TRITC-Lac-β-CyD was associated with NPC-like HepG2 cells higher than TRITC-β-CyD. In addition, TRITC-Lac-β-CyD was partially localized with endolysosomes after endocytosis. Thus, Lac-β-CyD entered NPC-like HepG2 cells via asialoglycoprotein receptor (ASGPR)-mediated endocytosis and decreased the accumulation of intracellular cholesterol in NPC-like HepG2 cells. These results suggest that Lac-β-CyD may have the potential as a drug for the treatment of hepatosplenomegaly in NPC disease.

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Multi-parameter analysis of combined hepatotoxicity induced by mycotoxin mixtures in HepG2 cells

World Mycotoxin Journal ◽

10.3920/wmj2017.2201 ◽

2018 ◽

Vol 11 (2) ◽

pp. 225-235 ◽

Cited By ~ 4

Author(s):

Y. Li ◽

T.Q. Wang ◽

J. Wu ◽

X.L. Zhang ◽

Y.Y. Xu ◽

...

Keyword(s):

Hepg2 Cells ◽

Transmembrane Potential ◽

Synergistic Effects ◽

Cell Loss ◽

Parameter Analysis ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Evaluation Models ◽

High Content Analysis ◽

Aflatoxin B

Evaluation of combined toxicity and exploring the corresponding mechanism is of great significance in characterising the interactions of mixed mycotoxins. This study used high content analysis and multiple evaluation models to estimate combined toxic hepatotoxicity in HepG2 cells, due to aflatoxin B1, zearalenone and deoxynivalenol, which are often detected simultaneously in the same grain sample. All mycotoxins induced cell loss in HepG2 cells in a concentration dependent manner. The combined toxic effects observed by multiple evaluation models (CA, IA and CI) suggested a similar mechanism and dominant synergistic effects for binary and ternary combinations. Based on reactive oxygen species, intracellular glutathione (GSH), and mitochondrial transmembrane potential (MMP) assessment, the synergistic mechanisms may be associated with mitochondrial damage by reducing GSH and MMP.

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Procyanidin B2 Alleviates Palmitic Acid-Induced Injury in HepG2 Cells via Endoplasmic Reticulum Stress Pathway

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/8920757 ◽

2021 ◽

Vol 2021 ◽

pp. 1-14

Author(s):

Yi-Ming Li ◽

Shao-Yang Zhao ◽

Huan-Huan Zhao ◽

Bao-Hua Wang ◽

Sai-Mei Li

Keyword(s):

Endoplasmic Reticulum ◽

Endoplasmic Reticulum Stress ◽

Er Stress ◽

Hepg2 Cells ◽

Cellular Injury ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Protein Levels ◽

Caspase 1 ◽

The Metabolic Syndrome

Nonalcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the metabolic syndrome featuring ectopic lipid accumulation in hepatocytes. NAFLD has been a severe threat to humans with a global prevalence of over 25% yet no approved drugs for the treatment to date. Previous studies showed that procyanidin B2 (PCB2), an active ingredient from herbal cinnamon, has an excellent hepatoprotective effect; however, the mechanism remains inconclusive. The present study aimed to investigate the protective effect and underlying mechanism of PCB2 on PA-induced cellular injury in human hepatoma HepG2 cells. Our results showed that PA-induced oxidative stress, calcium disequilibrium, and subsequent endoplasmic reticulum stress (ERS) mediated cellular injury, with elevated protein levels of GRP78, GRP94, CHOP, and hyperphosphorylation of PERK and IRE1α as well as the increased ratio of Bax/Bcl-2, which was restored by PCB2 in a concentration-dependent manner, proving the excellent antiapoptosis effect. In addition, 4-phenylbutyric acid (4-PBA), the ER stress inhibitor, increased cell viability and decreased protein levels of GRP78 and CHOP, which is similar to PCB2, and thapsigargin (TG), the ER stress agonist, exhibited conversely meanwhile partly counteracted the hepatic protection of PCB2. What is more, upregulated protein expression of p-IKKα/β, p-NF-κB p65, NLRP3, cleaved caspase 1, and mature IL-1β occurred in HepG2 cells in response to PA stress while rescued with the PCB2 intervention. In conclusion, our study demonstrated that PA induces ERS in HepG2 cells and subsequently activates downstream NLRP3 inflammasome-mediated cellular injury, while PCB2 inhibits NLRP3/caspase 1/IL-1β pathway, inflammation, and apoptosis with the presence of ERS, thereby promoting cell survival, which may provide pharmacological evidence for clinical approaches on NAFLD.

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Celastrol ameliorates acetaminophen-induced oxidative stress and cytotoxicity in HepG2 cells

Human & Experimental Toxicology ◽

10.1177/0960327117734622 ◽

2017 ◽

Vol 37 (7) ◽

pp. 742-751 ◽

Cited By ~ 9

Author(s):

AT Jannuzzi ◽

M Kara ◽

B Alpertunga

Keyword(s):

Oxidative Stress ◽

Cell Viability ◽

Hepg2 Cells ◽

Membrane Integrity ◽

Neutral Red ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Tripterygium Wilfordii Hook F ◽

Cell Membrane Integrity ◽

And Oxidative Stress

Acetaminophen (APAP) is the most commonly used analgesic and antipyretic drug in the world. However, hepatotoxicity caused by APAP overdose is the most frequent cause of acute liver failure worldwide and oxidative stress involved in the pathogenesis of APAP hepatotoxicity. Celastrol is a natural triterpenoid derived from Tripterygium wilfordii Hook F. that exhibits antioxidant, anti-inflammatory, and antitumor activities. In this study, we aimed to investigate the potential ameliorative effects of celastrol against APAP-induced cytotoxicity and oxidative stress. Human hepatocellular carcinoma cells (HepG2) were incubated with 20 mM of APAP for 24 h and posttreated with 50 nM, 100 nM, or 200 nM of celastrol for a further 24 h. The methylthiazolyldiphenyl-tetrazolium bromide, lactate dehydrogenase, and neutral red uptake assays showed celastrol posttreatments recovered cell viability and cell membrane integrity in a concentration-dependent manner. Celastrol posttreatments exerted a significant increase in the glutathione content and a decrease in the malondialdehyde and protein carbonylation levels. Also, celastrol posttreatments attenuated the APAP-induced oxidative stress by raising glutathione peroxidase, glutathione reductase, and catalase activities. However, superoxide dismutase activity did not change. In conclusion, celastrol treatment may improve cell viability and increase cellular antioxidant defense in HepG2 cells. These results suggest that celastrol may have the potential to ameliorate the APAP-induced oxidative stress and cytotoxicity.

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Different Induction Effects of Praziquantel Racemate and Enantiomers on the Enzyme CYP3A4 Mediated by Pregnane X Receptor and Its Variants

Current Drug Metabolism ◽

10.2174/1389200221999210104204057 ◽

2021 ◽

Vol 21 ◽

Author(s):

Ran Meng ◽

Xueli Zhang ◽

Haina Wang ◽

Danlu Zhang ◽

Xin Zhao

Keyword(s):

Hepg2 Cells ◽

Pregnane X Receptor ◽

Luciferase Reporter ◽

Dependent Manner ◽

Luciferase Reporter Gene ◽

Concentration Dependent Manner ◽

Protein Levels ◽

Mrna And Protein Expression ◽

Polymerase Chain ◽

Expression Plasmids

Background: Praziquantel (PZQ), which possesses an asymmetric center, is classified as a pyrazinoisoquinoline and has been the mainstay in the treatment of schistosomiasis since 1980. PZQ undergoes a pronounced first-pass metabolism in the liver through the CYP450 system which could be mediated by nuclear receptors. Objective: The purpose of this study was to investigate the possible different induction effects of CYP3A4 by PZQ racemate and enantiomers via the pregnane X receptor (PXR) and the effect of PXR polymorphism on the induction potency of PZQs. Methods: The dual-luciferase reporter gene systems constructed in HepG2 cells were used to measure the abilities of PZQs to induce CYP3A4 expression mediated by PXR. The mRNA and protein levels of CYP3A4 were evaluated by polymerase chain reaction (PCR) and western blotting, respectively. Results: In HepG2 cells transfected with PXRwt, PXR158, PXR163, PXR370 or PXR403 expression plasmids, PZQ racemate and its enantiomers up-regulated the luciferase activity in a concentration-dependent manner, while reaching saturation after transfected with PXR379 expression plasmids. The mRNA and protein expression of CYP3A4 was effectively activated in PXR-transfected HepG2 cells. The induction ability of CYP3A4 mediated by PXR activation by PZQ racemate and its enantiomers were statistically different between the same PXR group and different PXR groups. Conclusion: The enantioselective induction effects of PZQs on CYP3A4 were related to the enantioselective activations of PXR by PZQs and were influenced by the PXR gene polymorphism. These findings provide a basis for further understanding the enantiomeric metabolism and the variable efficacy of PZQs.

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Chalcone-thiosemicarbazone Hybrids as Inhibitors of Human Hepatocellular Carcinoma HepG2 Cells Viability and Oxygen Consumption

Current Bioactive Compounds ◽

10.2174/1573407218666220111104011 ◽

2022 ◽

Vol 18 ◽

Author(s):

Vivian Cordeiro Rodrigues ◽

William Queiroz Felippe ◽

Carla Marins Goulart ◽

Aurea Echevarria ◽

Ana Paula Pereira da Silva

Keyword(s):

Hepatocellular Carcinoma ◽

Oxygen Consumption ◽

Cell Viability ◽

Hepg2 Cells ◽

Atp Synthesis ◽

Human Hepatocellular Carcinoma ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Open Chain

Background: Chalcones are open-chain flavonoids especially attractive to medicinal chemistry due to their easy synthesis and the possibility of structural modifications. Objective: Evaluate the in vitro anticancer activity of a series of hybrids chalcones-thiosemicarbazones against the human hepatocellular carcinoma cell line, HepG2. Methods: Seven hybrid chalcones-thiosemicarbazones (CTs), 3-(4’-X-phenyl)-1-phenylprop-2-en-1-one thiosemicarbazone, where X=H (CT-H), CH3 (CT-CH3), NO2 (CT-NO2), Cl (CT-Cl), CN (CT-CN), F (CT-F) and Br (CT-Br), were synthesized and their effects on cells viability and mitochondrial oxygen consumption were accessed. Results: Incubation with CTs caused a decrease in HepG2 cells viability in a time-concentration-dependent manner. The most effective compounds in inhibiting cell viability, after 24 hours of treatment, were CT-Cl and CT-CH3 (IC50 20.9 and 23.63 μM, respectively). In addition, using 10 M and only 1 hour of pre-incubation, CT-CH3 caused a reduction in basal respiration (-37%), oxygen consumption coupled with ATP synthesis (-60%) and maximum oxygen consumption (-54%). These alterations in respiratory parameters may be involved with the inhibitory effects of CT-CH3, since significant changes in oxygen consumption rates were observed in a condition that anticipates more significant losses of cell viability. The ADME parameters and the no violation of Lipinski Rule of Five showed that all compounds are safe. Conclusion: These results may contribute to the knowledge about the effects of CTs on these cells and the development of new treatments against HCCs.

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Cyclopentadione-aniline conjugate suppresses proliferation and induces apoptosis in liver cancer cells via up-regulation of p38 phosphorylation

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v18i3.9 ◽

2021 ◽

Vol 18 (3) ◽

pp. 505-511

Author(s):

Heze Yu ◽

Geao Liang ◽

Bo Dou ◽

Jiangdong Ni

Keyword(s):

Liver Cancer ◽

Cancer Cells ◽

Hepg2 Cells ◽

Dependent Manner ◽

Transwell Assay ◽

Invasive Potential ◽

Concentration Dependent Manner ◽

Liver Cancer Cells ◽

Proliferation And Apoptosis

Purpose: To investigate the effect of cyclopentadione-aniline conjugate (CAC) on proliferation of liver cancer cells. Methods: MTT assay and flow cytometry were used for the determination of the effect of CAC on cell proliferation and apoptosis. Western blotting was used to measure the influence of CAC on the expressions of various proteins, while Matrigel-coated Transwell assay was used for assessment of cell invasion. Results: CAC inhibited proliferation of liver cancer cells in a concentration-dependent manner. The degree of proliferation of HepG2 cells was 98, 89, 76, 66, 51, 42 or 36 %, on treatment with 0.25, 0.5, 1.0, 1.5, 2.0, 2.5 or 3.0 µM CAC, respectively. In H4TG cells, treatment with 0.25, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 µM CAC decreased proliferation of cells to 99, 91, 79, 70, 54, 46 and 40 %, respectively. Apoptosis was induced in 34.56, 37.37 and 52.98 % cells, on treatment with 2.0, 2.5 and 3.0 µM CAC, respectively. The invasive potential of HepG2 cells was significantly decreased by CAC (p < 0.05). Marked decreases were observed in Bcl-2, MMP-2, MMP-9, c-ERK1/2 and phospho-Akt levels in CACtreated HepG2 cells. CAC treatment markedly upregulated Bax and phospho ERK1/2, but significantly downregulated phospho PI3K, phospho mTOR and phospho Akt in HepG2 cells (p < 0.05). However, the level of phospho p38 was decreased in CAC-treated cells. Conclusion: These results demonstrate that CAC inhibits proliferation of liver cancer cells via apoptosis induction. Thus, CAC can potentially be used for the treatment of liver cancer in humans

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EDTA reduces cadmium toxicity in mustard (Brassica juncea L.) by enhancing metal chelation, antioxidant defense and glyoxalase systems

Acta Agrobotanica ◽

10.5586/aa.1772 ◽

2019 ◽

Vol 72 (2) ◽

Cited By ~ 4

Author(s):

Jubayer Al Mahmud ◽

Mirza Hasanuzzaman ◽

Kamrun Nahar ◽

Anisur Rahman ◽

Masayuki Fujita

Keyword(s):

Lipid Peroxidation ◽

Brassica Juncea ◽

Antioxidant Defense ◽

Ethylenediaminetetraacetic Acid ◽

Cadmium Toxicity ◽

Monodehydroascorbate Reductase ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Cd Uptake ◽

Mg Content

To investigate the possible role of EDTA in mitigating cadmium (Cd) toxicity, we treated mustard (Brassica juncea L.) seedlings with CdCl2 (0.5 mM and 1.0 mM, 3 days) alone and in combination with 0.5 mM EDTA in a semihydroponic medium. In the absence of EDTA, mustard seedlings accumulated Cd in their roots and shoots in a concentration dependent manner. Overaccumulation of Cd boosted generation of hydrogen peroxide (H2O2) and superoxide anions (O2•−), increased lipoxygenase (LOX) activity, lipid peroxidation, and cytotoxic methylglyoxal (MG) content. It also disturbed components of the antioxidant defense and glyoxalase systems. Furthermore, Cd stress decreased growth, leaf relative water content (RWC) and chlorophyll (chl) content but augmented the proline (Pro) content. On the other hand, EDTA supplemented Cd-stressed seedlings improved the constituents of the AsA-GSH cycle with the upregulated activities of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT). Moreover, addition of EDTA to the Cd-stressed seedlings notably enhanced Gly I activity in contrast to the stress treatment. Ethylenediaminetetraacetic acid decreased Cd accumulation in the both shoots and roots, as well as increased other nonprotein thiols (NPTs) in leaves, including the phytochelatin (PC) content. It also decreased H2O2 and O2•− generation, lipid peroxidation and MG content but enhanced RWC, chl and Pro contents in the leaves, which confirmed the improved growth of seedlings. The findings of the study suggest that exogenous application of EDTA to the Cd-treated seedlings reduces Cd-induced oxidative injuries by restricting Cd uptake, increasing NPTs concentration and upregulating most of the components of their antioxidant defense and glyoxalase systems.

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The Effect of Icariin on Human Hepatoellular Carcinomas Cell Apoptosis and Cell Cycle Arrest

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2814 ◽

2021 ◽

Vol 11 (12) ◽

pp. 2389-2394

Author(s):

Zhenyu Feng ◽

Chao Chen ◽

Zhenyu Ye ◽

Jiaming Xie ◽

Wei Chen ◽

...

Keyword(s):

Cell Cycle ◽

Membrane Potential ◽

Liver Cancer ◽

Cell Cycle Arrest ◽

Cell Apoptosis ◽

Hepg2 Cells ◽

Mitochondrial Membrane ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Cycle Arrest

Background: Natural products often have novel frameworks and unique mechanisms, It is an important way to develop new anti-tumor drugs.The paper explored the effect of Icariin on HepG2 cell apoptosis and cell cycle arrest. Material and methods: Human liver cancer HepG2 cells were studied. The biological activity of Icariin on HepG2 cells was comprehensively investigated. The mechanism was preliminarily explored from the aspects of proliferation, cell cycle, and apoptosis. Results: Icariin showed significant inhibitory effect on cell proliferation after administration, and the effect was time-and-concentration-dependent. Annexin V-PI detection showed that, after 48 hours of administration of Icariin at different concentrations, the apoptosis rate of HepG2 cells increased in a concentration-dependent manner. The results of Hoechst 33342 staining showed that, after 48 hours of intervention with Icariin at different concentrations, HepG2 cells appeared densely stained and granular fluorescence, characterizing apoptosis. In the JC-1 mitochondrial membrane potential experiment, Icariin was found to destroy cell mitochondrial membrane potential and induce HepG2 cell apoptosis. After 48 h administration of Icariin at different concentrations, Bcl-2 and survivin proteins were down-regulated, and Bax was up-regulated, both in a concentration-dependent manner. PI single staining combined with flow cytometry to detect cell cycle results showed that, Icariin can induce G2/M phase arrest of HepG2 cells, and is time-and-concentration-dependent. Western Blot detection revealed that Icariin can down-regulate the cycle-related proteins Cyclin B and CDK1 in a concentration-dependent manner, and also significantly down-regulate the expressions of p-AKT, AKT, p-ERK and ERK proteins. Conclusion: Icariin is a selectively potential active compound that can treat liver cancer. The paper provided theoretical basis and experimental support for the clinical application of Icariin in the drug treatment of liver cancer. It is necessary to further study the antitumor effect of Icariin, explore and find the target, and provide higher selectivity for the treatment of liver cancer by Icariin.

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