Characterisation of some cytotoxic endpoints using rat liver and HepG2 spheroids as in vitro models and their application in hepatotoxicity studies. I. Glucose metabolism and enzyme release as cytotoxic markers

Xu, Jinyi; Ma, Mingwen; Purcell, Wendy M.

doi:10.1016/s0041-008x(03)00089-9

Cited by 70 publications

(49 citation statements)

References 53 publications

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“…These spheroids have a structure similar to actual liver tissues and can maintain a higher level of liver-specific functions over a longer period of time (Landry et al, 1985;Koide et al, 1990;Tong et al, 1992;Peshwa et al, 1996;Selden et al, 2000;Khalil et al, 2001;Abu-Absi et al, 2002;Depreter et al, 2002). Therefore, various applications have been proposed for these spheroid cultures, including bioartificial livers and liver tissue engineering as well as an in vitro model for studying drug metabolism and hepatotoxicity (Harries et al, 2001;Yamashita et al, 2001;Fukuda et al, 2003;Xu et al, 2003;Otsuka et al, 2004;Fukuda and Nakazawa 2005;Fukuda et al, 2006). However, it is difficult to control the diameter of spheroids grown in these cultures.…”

Section: Discussionmentioning

confidence: 99%

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Nakamura¹,

Kato²,

Aizawa³

et al. 2011

J. Toxicol. Sci.

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-The Nanoculture plate (NCP) is a recently developed plate which essentially consists of a textured surface with specific characteristics that induce spheroid formation: microfabrications with a micro-square pattern on the culture surface. The NCP can be used to generate uniform adhesive spheroids of cancer cell lines using conventional techniques without the need of any animal compounds. In this study, we assessed the performance of human hepatoma cell line HepG2 cells cultured with an NCP to evaluate the effects of the NCP on their hepatocyte-specific functions. The NCP facilitated the formation of three-dimensional (3D) HepG2 cell architecture. HepG2 cells cultured with an NCP exhibited enhanced mRNA expression levels of albumin and cytochrome P450 (CYP) enzymes compared to those cultured with a two-dimensional (2D) conventional plate. The expression levels of two specific liver-enriched transcription factors, hepatocyte nuclear factor 4α (HNF4α) and CCAAT/enhancer binding protein α (C/EBPα), were higher in HepG2 cells grown with the NCP than those in HepG2 cells grown with conventional plates before albumin and CYP enzymes expression levels were increased. The inducibility of CYP1A2 and CYP3A4 mRNA following exposure to inducers in HepG2 cells cultured with an NCP was comparable to that in HepG2 cells cultured with conventional plates, while the expression levels of CYP1A2 and CYP3A4 mRNA following exposure to inducers were higher when using an NCP than when using conventional plates. These results suggest that the use of an NCP enhances the hepatocyte-specific functions of HepG2 cells, such as drug-metabolizing enzyme expression, making the NCP/ HepG2 system a useful tool for evaluating drug metabolism in vitro.

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Section: Discussionmentioning

confidence: 99%

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Nakamura¹,

Kato²,

Aizawa³

et al. 2011

J. Toxicol. Sci.

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“…The absorbance was measured at 450 nm on a microplate reader (Tecan). Since there is an interference between APAP and LDH assay (Xu et al, 2003) only the treatments where APAP is used in the same dose are compared and results are presented as absorbance at 450 nm.…”

Section: Methodsmentioning

confidence: 99%

Lactobacillus fermentum Postbiotic-induced Autophagy as Potential Approach for Treatment of Acetaminophen Hepatotoxicity

et al. 2017

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The aim of this study was to investigate the potential of postbiotics originated from Lactobacillus fermentum BGHV110 strain (HV110) to counteract acetaminophen (APAP)-induced hepatotoxicity in HepG2 cells. This strain was selected according to its autophagy inducing potential, based on previous studies reporting protective role of autophagy in APAP caused cellular damage. Cell viability was assessed using MTT and LDH assays, while autophagy was monitored by qPCR analysis of BECN1, Atg5, p62/SQSTM1, and PINK1 mRNA expression and by Western blot analysis of p62/SQSTM1 and lipidated LC3 accumulation. Our results showed that detrimental effect of APAP on cell viability was suppressed in the presence of HV110 which was linked with increased conversion of LC3 protein and p62/SQSTM1 protein degradation. Additionally, higher p62/SQSTM1 and PINK1 mRNA transcription were noticed in cells co-treated with APAP/HV110, simultaneously. In conclusion, this study suggests that HV110 enhances activation of PINK1-dependent autophagy in HepG2 cells and its eventual co-supplementation with APAP could be potentially used for alleviation of hepatotoxic side effects caused by APAP overdose.

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“…28,42,43 However, liver is the primary organ and liver cells become resistant rst for metabolism of glucose, resulting in impaired glycogen synthesis and failure to suppress the production of glucose. 44 Hence, HepG2 cell lines were employed in the present work to investigate glucose uptake by treatment with luteolin and luteolin-manganese(II) complex, and to elucidate the hypoglycemic effect of them. As shown in Fig.…”

mentioning

confidence: 99%

Enhanced antioxidant activity, antibacterial activity and hypoglycemic effect of luteolin by complexation with manganese(ii) and its inhibition kinetics on xanthine oxidase

Dong

Yang

et al. 2017

RSC Adv.

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The present study aims to improve the biological activities of luteolin by complexation with manganese(II).UV-visible spectroscopy, infrared spectroscopy, thermogravimetric analysis and elemental analysis were adopted to assess the relevant interaction of luteolin and manganese(II) ions and the chelation sites. The antioxidant activity, hypoglycemic effect and antimicrobial activity of luteolin-manganese(II) complex with respect to its parent luteolin and the inhibition effect of which on xanthine oxidase were investigated and compared. The spectroscopic data indicated that luteolin reacts with manganese(II) cations through the chelation sites of 5-hydroxy and 4-carbonyl in two luteolin molecules. Antioxidant and antibacterial activity were enhanced after the complexation of manganese(II) cations with luteolin. An inhibition effect assay found that luteolin and luteolin-manganese(II) complex reversibly inhibited xanthine oxidase in a competitive manner. Luteolin-manganese(II) complex had a more remarkable hypoglycemic effect than luteolin by increasing the glucose consumption in liver tissue.

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Characterisation of some cytotoxic endpoints using rat liver and HepG2 spheroids as in vitro models and their application in hepatotoxicity studies. I. Glucose metabolism and enzyme release as cytotoxic markers

Cited by 70 publications

References 53 publications

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Lactobacillus fermentum Postbiotic-induced Autophagy as Potential Approach for Treatment of Acetaminophen Hepatotoxicity

Enhanced antioxidant activity, antibacterial activity and hypoglycemic effect of luteolin by complexation with manganese(ii) and its inhibition kinetics on xanthine oxidase

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