Global Proteomics to Study Silica Nanoparticle-Induced Cytotoxicity and Its Mechanisms in HepG2 Cells

Silica nanoparticles (SiO2 NPs) are commonly used in medical and pharmaceutical fields. Research into the cytotoxicity and overall proteomic changes occurring during initial exposure to SiO2 NPs is limited. We investigated the mechanism of toxicity in human liver cells according to exposure time [0, 4, 10, and 16 h (h)] to SiO2 NPs through proteomic analysis using mass spectrometry. SiO2 NP-induced cytotoxicity through various pathways in HepG2 cells. Interestingly, when cells were exposed to SiO2 NPs for 4 h, the morphology of the cells remained intact, while the expression of proteins involved in mRNA splicing, cell cycle, and mitochondrial function was significantly downregulated. These results show that the toxicity of the nanoparticles affects protein expression even if there is no change in cell morphology at the beginning of exposure to SiO2 NPs. The levels of reactive oxygen species changed significantly after 10 h of exposure to SiO2 NPs, and the expression of proteins associated with oxidative phosphorylation, as well as the immune system, was upregulated. Eventually, these changes in protein expression induced HepG2 cell death. This study provides insights into cytotoxicity evaluation at early stages of exposure to SiO2 NPs through in vitro experiments.

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AAnti-Inflammatory Effects of Plasma Circulating Exosomes Obtained from Normal-Weight and Obese Subjects on Hepatocytes

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530320666200505121426 ◽

2020 ◽

Vol 20 ◽

Author(s):

Reza Afrisham ◽

Sahar Sadegh-Nejadi ◽

Reza Meshkani ◽

Solaleh Emamgholipour ◽

Molood Bagherieh ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Hepg2 Cells ◽

Human Liver ◽

Liver Cells ◽

Normal Weight ◽

Protein Levels ◽

Human Liver Cells ◽

Tnf Α ◽

Anti Inflammatory

Introduction: Obesity is a disorder with low-grade chronic inflammation that plays a key role in the hepatic inflammation and steatosis. Moreover, there are studies to support the role of exosomes in the cellular communications, the regulation of metabolic homeostasis and immunomodulatory activity. Accordingly, we aimed to evaluate the influence of plasma circulating exosomes derived from females with normal-weight and obesity on the secretion of inflammatory cytokines in human liver cells. Methods: Plasma circulating exosomes were isolated from four normal (N-Exo) and four obese (O-Exo) women. The exosomes were characterized and approved for CD63 expression (common exosomal protein marker) and morphology/size using the western blot and TEM methods, respectively. The exosomes were used for stimulation of HepG2 cells in vitro. After 24 h incubation, the protein levels of TNF-α,IL-6, and IL-1β were measured in the culture supernatant of HepG2 cells using the ELISA kit. Results: The protein levels of IL-6 and TNF-α in the cells treated with O-Exo and N-Exo reduced significantly in comparison with control group (P=0.039 and P<0.001 respectively), while significance differences were not found between normal and obese groups (P=0.808, and P=0.978 respectively). However, no significant differences were found between three groups in term of IL-1β levels (P=0.069). Based on the correlation analysis, the protein levels of IL-6 were positively correlated with TNF-α (r 0.978, P<0.001). Conclusion: These findings suggest that plasma circulating exosomes have probably anti-inflammatory properties independently from body mass index and may decrease the secretion of inflammatory cytokines in liver. However, further investigations in vitro and in vivo are needed to address the anti-inflammatory function of N-Exo and O-Exo in human liver cells and/or other cells.

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Organophosphorus Flame Retardant TDCPP Displays Genotoxic and Carcinogenic Risks in Human Liver Cells

Cells ◽

10.3390/cells11020195 ◽

2022 ◽

Vol 11 (2) ◽

pp. 195

Author(s):

Quaiser Saquib ◽

Abdullah M. Al-Salem ◽

Maqsood A. Siddiqui ◽

Sabiha M. Ansari ◽

Xiaowei Zhang ◽

...

Keyword(s):

Flame Retardant ◽

Hepg2 Cells ◽

Human Liver ◽

Human Cancer ◽

Liver Cells ◽

Consumer Products ◽

In Vivo Test ◽

Human Liver Cells

Tris(1,3-Dichloro-2-propyl)phosphate (TDCPP) is an organophosphorus flame retardant (OPFR) widely used in a variety of consumer products (plastics, furniture, paints, foams, and electronics). Scientific evidence has affirmed the toxicological effects of TDCPP in in vitro and in vivo test models; however, its genotoxicity and carcinogenic effects in human cells are still obscure. Herein, we present genotoxic and carcinogenic properties of TDCPP in human liver cells (HepG2). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and neutral red uptake (NRU) assays demonstrated survival reduction in HepG2 cells after 3 days of exposure at higher concentrations (100–400 μM) of TDCPP. Comet assay and flow cytometric cell cycle experiments showed DNA damage and apoptosis in HepG2 cells after 3 days of TDCPP exposure. TDCPP treatment incremented the intracellular reactive oxygen species (ROS), nitric oxide (NO), Ca2+ influx, and esterase level in exposed cells. HepG2 mitochondrial membrane potential (ΔΨm) significantly declined and cytoplasmic localization of P53, caspase 3, and caspase 9 increased after TDCPP exposure. qPCR array quantification of the human cancer pathway revealed the upregulation of 11 genes and downregulation of two genes in TDCPP-exposed HepG2 cells. Overall, this is the first study to explicitly validate the fact that TDCPP bears the genotoxic, hepatotoxic, and carcinogenic potential, which may jeopardize human health.

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The effect of liver donors’ age, gender and metabolic state on pancreatic lineage activation

Regenerative Medicine ◽

10.2217/rme-2020-0092 ◽

2021 ◽

Vol 16 (1) ◽

pp. 19-31

Author(s):

Ioan V Matei ◽

Irit Meivar-Levy ◽

Daniela Lixandru ◽

Simona Dima ◽

Ioana R Florea ◽

...

Keyword(s):

Replacement Therapy ◽

Human Liver ◽

Liver Cells ◽

Diabetic Patients ◽

Metabolic State ◽

Human Liver Cells ◽

Metabolic States ◽

Different Ages ◽

Liver Donors

Autologous cells replacement therapy by liver to pancreas transdifferentiation (TD) allows diabetic patients to be also the donors of their own therapeutic tissue. Aim: To analyze whether the efficiency of the process is affected by liver donors’ heterogeneity with regard to age, gender and the metabolic state. Materials & methods: TD of liver cells derived from nondiabetic and diabetic donors at different ages was characterized at molecular and cellular levels, in vitro. Results: Neither liver cells proliferation nor the propagated cells TD efficiency directly correlate with the age (3–60 years), gender or the metabolic state of the donors. Conclusion: Human liver cells derived from a wide array of ages and metabolic states can be used for autologous cells therapies for diabetics.

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Microcystins activate nuclear factor erythroid 2-related factor 2 (Nrf2) in human liver cells in vitro – Implications for an oxidative stress induction by microcystins

Toxicon ◽

10.1016/j.toxicon.2016.12.012 ◽

2017 ◽

Vol 126 ◽

pp. 47-50 ◽

Cited By ~ 9

Author(s):

Johan Lundqvist ◽

Heidi Pekar ◽

Agneta Oskarsson

Keyword(s):

Oxidative Stress ◽

Human Liver ◽

Liver Cells ◽

Related Factor ◽

Nuclear Factor ◽

Stress Induction ◽

Human Liver Cells

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Qizhi Jiangtang Jiaonang Improves Insulin Signaling and Reduces Inflammatory Cytokine Secretion and Reactive Oxygen Species Formation in Insulin Resistant HepG2 Cells

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/518639 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8

Author(s):

Xiao-Tian Zhang ◽

Chun-Jiang Yu ◽

Jian-Wei Liu ◽

Yan-Ping Zhang ◽

Chao Zhang ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Palmitic Acid ◽

Hepg2 Cells ◽

Reactive Oxygen ◽

Glucose Consumption ◽

Control Group ◽

High Dose ◽

Oxygen Species ◽

Insulin Resistant

We analyzed the effects of a traditional Chinese medicine, Qizhi Jiangtang Jiaonang (QJJ), on insulin resistance (IR) in vitro. After an in vitro model of IR was established by treating human liver cancer cells (HepG2 cells) with palmitic acid, the cells were then treated with various concentrations of QJJ. Treatment with 400 µM palmitic acid for 24 h induced IR in HepG2 cells. The survival rate for HepG2 cells in the IR group was significantly lower than that of the untreated control group (P< 0.001); however, QJJ restored HepG2 cell survival (P< 0.001). As compared with HepG2 cells in the IR group, QJJ at all doses analyzed significantly increased glucose consumption (allP< 0.05). Moreover, treatment with all the QJJ doses significantly reduced the mean intracellular reactive oxygen species levels as compared with the IR group (allP< 0.05). Furthermore, high-dose QJJ reduced both TNF-αand IL-6 levels as compared to the IR group (allP< 0.05). QJJ ameliorated the altered PI3K, GLUT4, and RAGE expression observed with IR. In conclusion, QJJ can improve IR in HepG2 cells, which may be mediated through the IRS-1/PI3K/GLUT4 signaling pathway as well as regulation of NF-κB-mediated inflammation and oxidative stress.

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Potential Simultaneous Inhibitors of Angiotensin-Converting Enzyme 2 and Transmembrane Protease, Serine 2

Frontiers in Pharmacology ◽

10.3389/fphar.2020.584158 ◽

2020 ◽

Vol 11 ◽

Author(s):

Ching-Yuan Wu ◽

Yu-Shih Lin ◽

Yao-Hsu Yang ◽

Li-Hsin Shu ◽

Yu-Ching Cheng ◽

...

Keyword(s):

Protein Expression ◽

Hepg2 Cells ◽

Kidney Tissue ◽

Herbal Formula ◽

Angiotensin Converting Enzyme 2 ◽

Mrna And Protein Expression ◽

Almost All ◽

Lung And Kidney

Outbreak of coronavirus disease 2019 occurred in Wuhan and has rapidly spread to almost all parts of world. GB-1, the herbal formula from Tian Shang Sheng Mu of Chiayi Puzi Peitian Temple, is used for the prophylaxis of SARS-CoV-2 in Taiwan. In this study, we investigated that the effect of GB-1 and the index compounds of GB-1 on the ACE2 and TMPRSS2 expression through in vitro and in vivo study. In our result, GB-1 can inhibit ACE2 and TMPRSS2 protein expression in HepG2 cells, 293T cells, and Caco-2 cells without cytotoxicity. For the mouse model, GB-1 treatment could decrease ACE2 and TMPRSS2 expression levels of the lung and kidney tissue without adverse effects, including nephrotoxicity and hepatotoxicity. In the compositions of GB-1, 0.5–1 mg/ml of Glycyrrhiza uralensis Fisch. ex DC. extract could not inhibit ACE2 mRNA and protein expression in HepG2 cells. In addition, theaflavin-3-gallate could inhibit protein expression of ACE2 and TMPRSS2 without significant cytotoxicity. Our results suggest that GB-1 and theaflavin-3-gallate could act as potential candidates for prophylaxis or treatment of SARS-CoV-2 infection through inhibiting protein expression of ACE2 and TMPRSS2 for the further study.

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Protective effects of papaya extracts on tert-butyl hydroperoxide mediated oxidative injury to human liver cells (An in-vitro study)

Free Radicals and Antioxidants ◽

10.5530/ax.2012.3.2 ◽

2012 ◽

Vol 2 (3) ◽

pp. 10-19 ◽

Cited By ~ 6

Author(s):

Sheri-Ann Tan ◽

Sonia Ramos ◽

María Angeles Martin ◽

Raquel Mateos ◽

Michael Harvey ◽

...

Keyword(s):

Human Liver ◽

In Vitro Study ◽

Oxidative Injury ◽

Liver Cells ◽

Vitro Study ◽

Protective Effects ◽

Butyl Hydroperoxide ◽

Human Liver Cells ◽

Tert Butyl Hydroperoxide

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In Vitro Antihepatoma Activity of Novel 3D-Copper Cyanide Supramolecular Coordination Polymers

Journal of AOAC International ◽

10.5740/jaoacint.16-0141 ◽

2016 ◽

Vol 99 (5) ◽

pp. 1240-1246

Author(s):

Noura M Darwish ◽

Ahmed S Sultan ◽

Ahmed M Malki ◽

Hossam Khamis ◽

Mohamed El-Ziady

Keyword(s):

Protein Expression ◽

Western Blot ◽

Coordination Polymers ◽

Hepg2 Cells ◽

Inhibitory Effect ◽

Dependent Manner ◽

Invasive Potential ◽

Dna Ladder ◽

Supramolecular Coordination

Abstract This study aimed to investigate the inhibitory effect of novel 3D-organocopper supramolecular coordination polymers (SCPs) on the invasive potential of HepG2 cells. Chemoprevention could represent an important means to inhibit the process of hepatocarcinogenesis. The inhibitory effect of an SCP compound on the proliferation of HepG2 hepatoma cells was evaluated by cell vibility assay. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of the SCP compound on phosphorylated ERK1/2, Bcl-2, and β-catenin protein expression of HepG2 cells was analyzed by Western blot. The SCP compound exerted an inhibitory effect on HepG2 cell proliferation in a dose-dependent manner. This inhibitory effect was confirmed by examination of cell morphology and DNA fragmentation. Furthermore, Western blot analysis revealed that phosphorylated ERK1/2 and β-catenin protein expression was inhibited after 24 h of treatment with the SCP compound, and that this event was associated with decreased Bcl-2 expression. We concluded that SCP can effectively inhibit the invasive potential of the ERK signaling pathway in HepG2 cells by altering apoptosis and by inhibiting Bcl-2 and β-catenin, which may play a significant role in this process.

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