Increased susceptibility of fat-laden Zucker-rat hepatocytes to bile acid-induced oncotic necrosis: An in vitro model of steatocholestasis

2005 ◽  
Vol 145 (5) ◽  
pp. 247-262 ◽  
Author(s):  
Gregory E. Kobak ◽  
Rolf Dahl ◽  
Michael W. Devereaux ◽  
Eric Gumpricht ◽  
Maret Traber ◽  
...  
Keyword(s):  
Bile Acid ◽  
In Vitro Model ◽  
Rat Hepatocytes ◽  
Zucker Rat ◽  
Vitro Model ◽  
Increased Susceptibility

An In Vivo/In Vitro Study of Allyl Alcohol Toxicity Using Enzyme Inhibitors

1993 ◽  
Vol 21 (1) ◽  
pp. 38-42
Author(s):  
Romana Pulci ◽  
Donatella Moneta ◽  
Philippe Dostert ◽  
Marco Brughera ◽  
Giovanna Scampini ◽  
...  
Keyword(s):  
Aldehyde Dehydrogenase ◽  
Allyl Alcohol ◽  
In Vitro Model ◽  
Enzyme Inhibitors ◽  
In Vitro Study ◽  
Rat Hepatocytes ◽  
Isolated Rat Hepatocytes ◽  
Vitro Model

The aim of this study was to verify an in vitro model of hepatotoxicity, designed to assess the production of reactive species from biologically-inert chemicals through their metabolic transformation. One example is allyl alcohol, which produces acrolein through the action of the enzyme alcohol dehydrogenase. Acrolein is a highly hepatotoxic aldehyde which is detoxified to acrylic acid by aldehyde dehydrogenase (ALDH). A deficiency of this enzyme, common in some Asian populations, can give rise to pathological conditions of hepatotoxicity. Isolated rat hepatocytes were incubated with allyl alcohol with and without cyanamide, a known inhibitor of ALDH. The toxicity of allyl alcohol, assessed on the basis of release of glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT) and lactate dehydrogenase (LDH) into the culture medium, was dramatically increased by the addition of cyanamide. In vivo, the same treatment scheme was used in rats treated with allyl alcohol with or without cyanamide pretreatment. It was also demonstrated that allyl alcohol toxicity is dramatically enhanced by the addition of an aldehyde dehydrogenase inhibitor, as shown by plasma levels of hepatic enzymes (GOT, GPT and LDH) and by histological findings. We believe that this in vitro model, involving the use of enzyme inhibitors, could be useful for verification of the hypothesis that hepatotoxins, such as acrolein, are produced from some pharmaceutical and other chemical compounds.

scholarly journals Development of a Rat Sandwich-Cultured Hepatocytes Model Expressing Functional Human Organic Anion Transporting Polypeptide (OATP) 1B3: A Potential Screening Tool for Liver-Targeting Compounds

2021 ◽  
Vol 24 ◽  
pp. 475-483
Author(s):  
Taleah Farasyn ◽  
Chao Xu ◽  
Wei Yue
Keyword(s):  
Biliary Excretion ◽  
In Vitro Model ◽  
Organic Anion ◽  
Rat Hepatocytes ◽  
Drug Uptake ◽  
Liver Targeting ◽  
Organic Anion Transporting Polypeptide ◽  
Vitro Model ◽  
In Cells

Purpose: Organic anion transporting polypeptide (OATP) 1B3 transports many clinically important drugs, including statins, from blood into the liver. It exclusively expresses in human liver under normal physiological conditions. There is no rodent ortholog of human OATP1B3. Tissue targeting of therapeutic molecules mediated by transporters, including liver-targeting via liver-specific OATPs, is an emerging area in drug development. Sandwich-cultured primary hepatocytes (SCH) are a well characterized in vitro model for assessment of hepatic drug uptake and biliary excretion. The current study was designed to develop a novel rat SCH model expressing human OATP1B3 to study the hepatic disposition of OATP1B3 substrates. Methods: Primary rat hepatocytes transduced with adenoviral vectors expressing FLAG-tagged OATP1B3 (Ad-OATP1B3), a control vector Ad-LacZ, or that were non-transduced were cultured in a sandwich configuration. FLAG immunoblot and immunofluorescence-staining determined expression and localization of OATP1B3. Uptake of [3H]-cholecystokinin octapeptide (CCK-8), a specific OATP1B3 substrate, was determined. Taurocholate (TC) is a substrate routinely used in SCH to assess biliary excretion via bile canaliculi (BC) and is also a substrate of OATP1B3. [3H]-TC accumulation in cells+BC, cells, biliary excretion index (BEI) and in vitro Clbiliary were determined using B-CLEAR® technology. Results: OATP1B3 protein was extensively expressed and primarily localized on the plasma membrane in day 4 Ad-OATP1B3-transduced rat SCH. [3H]-CCK-8 accumulation in cells+BC was significantly greater (~5-13 folds, p<0.001) in day 4 SCH with vs. without Ad-OATP1B3-transduction. Expressing OATP1B3 in rat SCH significantly increased [3H]-TC accumulation in cells+BC and cells, without affecting BEI and in vitro Clbiliary. Conclusions: Rat SCH expressing human OATP1B3-is a novel in vitro model allowing simultaneous assessment of hepatic uptake, hepatocellular accumulation and biliary excretion process of a human OATP1B3 substrate. This model could be a potential tool for screening for liver-targeting compounds mediated by OATP1B3.

Bile acid synthesis in isolated rat hepatocytes

1978 ◽  
Vol 56 (8) ◽  
pp. 780-783 ◽  
Author(s):  
I. M. Yousef ◽  
J. Ho ◽  
K. N. Jeejeebhoy
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Rat Hepatocytes ◽  
Acid Synthesis ◽  
Isolated Hepatocytes ◽  
Bile Acid Synthesis ◽  
Isolated Rat Hepatocytes ◽  
Adult Rat ◽  
Vitro Model

Normal adult rat hepatocytes were incubated for 48 h and the concentration of total and individual bile acids in homogenized samples of the culture was measured at intervals during the incubation, using radiogas chromatography and isotope derivative assay. The net increase in bile acids over the value observed at the start of the culture was taken as synthesis. The results showed that bile acid synthesis was linear up to 24 h of incubation, at a rate of 20 nmol/g hepatocytes per hour, and that 85% of the newly synthesized bile acid was cholic acid. The bile acid synthesized was mainly conjugated with taurine. These results suggest that isolated hepatocytes cultured in the way described could be a useful in vitro model for the study of bile acid synthesis.

Sandwich-cultured rat hepatocytes as an in vitro model to study canalicular transport alterations in cholestasis

2014 ◽  
Vol 89 (6) ◽  
pp. 979-990 ◽  
Author(s):  
Gisel S. Miszczuk ◽  
Ismael R. Barosso ◽  
Andrés E. Zucchetti ◽  
Andrea C. Boaglio ◽  
José M. Pellegrino ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Rat Hepatocytes ◽  
Cultured Rat Hepatocytes ◽  
Vitro Model

Steatosis sensitizes hepatocytes to bile acid-induced apoptosis in a human in vitro model

2008 ◽  
Vol 46 (01) ◽  
Author(s):  
T Pusl ◽  
N Wild ◽  
R Wimmer ◽  
T Vennegeerts ◽  
C Rust
Keyword(s):  
Bile Acid ◽  
In Vitro Model ◽  
Vitro Model ◽  
Induced Apoptosis

An in vitro model for investigation of vitamin A effects on wound healing

2021 ◽  
pp. 1-6
Author(s):  
Hoda Keshmiri Neghab ◽  
Mohammad Hasan Soheilifar ◽  
Gholamreza Esmaeeli Djavid
Keyword(s):  
Wound Healing ◽  
Endothelial Cell ◽  
Vitamin A ◽  
In Vitro Model ◽  
Cellular Proliferation ◽  
Endothelial Cell Migration ◽  
Normal Fibroblast ◽  
Anti Inflammatory ◽  
Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inflammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inflammation responses.

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