Evaluating the Effects of Dithiothreitol and Fructose on Cell Viability and Function of Cryopreserved Primary Rat Hepatocytes and HepG2 Cell Line

Abstract Objective In present study, the effects of the leaf extract of Pyrus biossieriana Buhse on tert-Butyl hydroperoxide (t-BHP) induced toxicity in the HepG2 cell line were investigated. Results HepG2 cells were exposed to different concentrations of both extract (1.5, 2.0, and 2.5 mg/mL) and t-BHP (100, 150, and 200 μM). The total flavonoid and phenolic contents, the cell viability, lipid peroxidation, NO generation, and the total antioxidant capacity in cell media were assessed. The amount of arbutin was estimated 12.6% of the dry weight of leaves (equivalent to 126 mg/g). Additionally, the amounts of flavonoids and phenols in extract were estimated 119 mg/g and 418 mg/g, respectively. The cells incubated with t-BHP showed a significant decrease in survival (p < 0.001). Preincubation with extract (1.5 mg/mL and 2.0 mg/mL) attenuated the t-BHP toxicity and increased the cell viability in cells exposed even to the highest concentration of t-BHP (200 μM) (p value < 0.001, and p value = 0.035) respectively. Additionally, treatment with extract reduced the cell growth suppression caused by t-BHP. The P. biossieriana Buhse leaf extract at concentrations of 1.5 and 2.0 mg/mL is capable of attenuating t-BHP-induced cytotoxicity in HepG2 cells.

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Curcumin Protects HepG2 Cells from the Cytotoxic Effect of Drugs with Anti-fibrotic Activity

10.21203/rs.3.rs-289436/v1 ◽

2021 ◽

Author(s):

Mariana Y. Medina-Pizaño ◽

Marina N. Medina-Rosales ◽

Esperanza Sánchez-Alemán ◽

Sandra L. Martínez-Hernández ◽

Liseth R. Aldaba-Muruato ◽

...

Keyword(s):

Cell Viability ◽

Cell Line ◽

Hepg2 Cell ◽

Hepg2 Cells ◽

Morphological Changes ◽

Liver Parenchyma ◽

Hepg2 Cell Line ◽

Secondary Effects ◽

Hamster Model ◽

Hematoxylin And Eosin

Abstract Background: The α and β adrenoblockers have been tested as an alternative treatment for chronic liver lesions such as fibrosis and cirrhosis in animal models, as well as their possible participation during the regeneration of the damage caused by liver cirrhosis in a hamster model. However, it was observed that doxazosin caused slight morphological changes in hepatocytes, while that curcumin showed protection to the hepatic parenchyma. Regardless, the pharmacokinetic effects of these 𝛼/𝛽 adrenoblockers on the hepatocytes' cell viability, possibly involved in the hepatic parenchyma's repopulation during cirrhosis reversal, are unknown. The present study aimed to elucidate the protective effect of curcumin on the possible side effects of doxazosin, tamsulosin, and carvedilol on the HepG2 cell line, drugs already tested with antifibrotic activity.Methods: HepG2 cells were exposed to 0.1, 0.5, 10, and 25 µM of doxazosin, carvedilol, and tamsulosin for 24, 48, and 72 h, for curcumin, cells were pretreated with 1 µM for 1 h before exposure to α and β adrenoblockers. The cell viability was assessed by MTT assay. The morphological changes were determined using hematoxylin and eosin (H&E) staining, scanning electron microscope (SEM), and acridine orange (AO) staining. Results: We observed that the doxazosin decreases cell viability dependently time and dose; carvedilol and tamsulosin increase cell proliferation. However, curcumin induces regulation of these effects in HepG2 cell line, increasing or maintaining viability compared to control. The pretreatment with curcumin regulated AST levels (aspartate aminotransferase) and ALT (alanine aminotransferase) in cells exposed to α and β adrenoblockers. The SEM and H&E staining provided evidence that doxazosin, carvedilol, and tamsulosin induced morphological changes in HepG2 cell line, depending on time and dose, approximately 80% of the cells treated with drugs were balonized, and curcumin protected these effects, maintaining the morphology in 90% of the treated cells.Conclusions: The present study demonstrates that curcumin protected the HepG2 cells against cytotoxicity and morphological changes induced by the α and β adrenoblockers attenuating secondary effects for possible oxidative stress. In this way, it is concluded that these treatments with antifibrotic effect, in co-treatment with curcumin, will not affect the possible repopulation process of the liver parenchyma during the reversion of fibrosis.

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3-Iodothyronamine and 3,5,3′-triiodo-L-thyronine reduce SIRT1 protein expression in the HepG2 cell line

Hormone Molecular Biology and Clinical Investigation ◽

10.1515/hmbci-2019-0045 ◽

2020 ◽

Vol 41 (1) ◽

Author(s):

Ginevra Sacripanti ◽

Leonardo Lorenzini ◽

Lavinia Bandini ◽

Sabina Frascarelli ◽

Riccardo Zucchi ◽

...

Keyword(s):

Protein Expression ◽

Cell Viability ◽

Cell Line ◽

Potential Role ◽

Colorimetric Assay ◽

Tumor Cell Line ◽

Rat Hepatocytes ◽

Carcinoma Cells ◽

Primary Rat Hepatocytes

AbstractBackground3-Iodothyronamine (T1AM) is an endogenous messenger chemically related to thyroid hormone. Recent results indicate significant transcriptional effects of chronic T1AM administration involving the protein family of sirtuins, which regulate important metabolic pathways and tumor progression. Therefore, the aim of this work was to compare the effect of exogenous T1AM and 3,5,3′-triiodo-L-thyronine (T3) chronic treatment on mammalian sirtuin expression in hepatocellular carcinoma cells (HepG2) and in primary rat hepatocytes at micromolar concentrations.Materials and methodsSirtuin (SIRT) activity and expression were determined using a colorimetric assay and Western blot analysis, respectively, in cells treated for 24 h with 1–20 μM T1AM or T3. In addition, cell viability was evaluated by the MTTtest upon 24 h of treatment with 0.1–20 μM T1AM or T3.ResultsIn HepG2, T1AM significantly reduced SIRT 1 (20 μM) and SIRT4 (10–20 μM) protein expression, while T3 strongly decreased the expression of SIRT1 (20 μM) and SIRT2 (any tested concentration). In primary rat hepatocytes, T3 decreased SIRT2 expression and cellular nicotinamide adenine dinucleotide (NAD) concentration, while on sirtuin activity it showed opposite effects, depending on the evaluated cell fraction. The extent of MTT staining was moderately but significantly reduced by T1AM, particularly in HepG2 cells, whereas T3 reduced cell viability only in the tumor cell line.ConclusionsT1AM and T3 downregulated the expression of sirtuins, mainly SIRT1, in hepatocytes, albeit in different ways. Differences in mechanisms are only observational, and further investigations are required to highlight the potential role of T1AM and T3 in modulating sirtuin expression and, therefore, in regulating cell cycle or tumorigenesis.

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Hepatoprotective activity of Flacourtia jangomas (Lour.) Raeuschleaves and fruit methanolic extract on paracetamol-induced hepatotoxicity in HepG2 Cells

Biomedicine ◽

10.51248/.v41i3.1197 ◽

2021 ◽

Vol 41 (3) ◽

pp. 587-591

Author(s):

Akshaya Pai ◽

Chandrakala Shenoy

Keyword(s):

Cell Viability ◽

Cell Line ◽

Hepg2 Cell ◽

Hepg2 Cells ◽

Methanolic Extract ◽

Hepatoprotective Activity ◽

Negative Control ◽

Hepg2 Cell Line ◽

Fruit Extract ◽

Deciduous Fruit

Introduction and Aim: Plants have become the current focus of research in treating the various diseases and ailments. Flacourtia jangomas (Lour.) Raeusch belongs to the familySalicaceae. Itis a small deciduous fruit tree having immense nutritional and medicinal significance. Different parts of the plant are pharmaceutically used forcuring various ailments. In this study, we investigated the hepatoprotective activity of Flacourtia jangomas (Lour.) Raeusch leaves and fruit methanolic extract on Paracetamol induced HepG2 cell line. Methods: The cytotoxic and hepatoprotective properties were evaluated by measuring cell viability; activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH); lipid peroxidation (malondialdehyde (MDA) levels). Results:The increased cell viability of 140.43± 4.07% and 133.93±3.20%was observed in HepG2 cells treated with methanolic extract of F. jangomas leaf and fruit extract respectively at 10µg/ml concentration and then decreased along with the rise of F. jangomas leaf and fruit extract concentrations. The level of LDH, ALT, AST and MDA decreased after F. jangomas leaf and fruit treatment compared to negative control. Conclusion: This study suggests that the methanolic Extract of F. jangomas (Lour.) Raeusch leaves(FJL)and fruit (FJF) shows hepatoprotective activity in Paracetamol induced HepG2 cell line by the decrease in AST and ALT activities and LDH and MDA level. Hence, it could be considered as a therapeutic agent in curing liver-related diseases.

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