Blood cell oxidative stress precedes hemolysis in whole blood-liver slice co-cultures of rat, dog, and human tissues

Journal Article · Sat May 01 00:00:00 EDT 2010 · Toxicology and Applied Pharmacology

DOI:https://doi.org/10.1016/j.taap.2010.01.017· OSTI ID:21344938

Sinclair, John R ^[1]; Fisher, Robyn L ^[2]; Morris, Stephen R; Way, William ^[1]

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1. Drug Safety Evaluation, Allergan, 2525 Dupont Dr, Irvine, CA 92612 (United States)
2. Vitron, Tucson, AZ (United States)

A novel in vitro model to investigate time-dependent and concentration-dependent responses in blood cells and hemolytic events is studied for rat, dog, and human tissues. Whole blood is co-cultured with a precision-cut liver slice. Methimazole (MMI) was selected as a reference compound, since metabolism of its imidazole thione moiety is linked with hematologic disorders and hepatotoxicity. An oxidative stress response occurred in all three species, marked by a decline in blood GSH levels by 24 h that progressed, and preceded hemolysis, which occurred at high MMI concentrations in the presence of a liver slice with rat (>= 1000 muM at 48 h) and human tissues (>= 1000 muM at 48 h, >= 750 muM at 72 h) but not dog. Human blood-only cultures exhibited a decline of GSH levels but minimal to no hemolysis. The up-regulation of liver genes for heme degradation (Hmox1 and Prdx1), iron cellular transport (Slc40a1), and GSH synthesis and utilization (mGST1 and Gclc) were early markers of the oxidative stress response. The up-regulation of the Kupffer cell lectin Lgals3 gene expression indicated a response to damaged red blood cells, and Hp (haptoglobin) up-regulation is indicative of increased hemoglobin uptake. Up-regulation of liver IL-6 and IL-8 gene expression suggested an activation of an inflammatory response by liver endothelial cells. In summary, MMI exposure led to an oxidative stress response in blood cells, and an up-regulation of liver genes involved with oxidative stress and heme homeostasis, which was clearly separate and preceded frank hemolysis.

• OSTI ID: 21344938
• Journal Information: Toxicology and Applied Pharmacology, Vol. 244, Issue 3; Other Information: DOI: 10.1016/j.taap.2010.01.017; PII: S0041-008X(10)00042-6; Copyright (c) 2010 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; ISSN 0041-008X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
BLOOD CELLS
DOGS
GENES
HEME
HEMOGLOBIN
HEMOLYSIS
HOMEOSTASIS
IN VITRO
INFLAMMATION
IRON
LIVER
MEN
METABOLISM
OXIDATION
RATS
RETICULOENDOTHELIAL SYSTEM
STRESSES
SYNTHESIS
TIME DEPENDENCE
ANIMAL TISSUES
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BODY
BODY FLUIDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
DECOMPOSITION
DIGESTIVE SYSTEM
ELEMENTS
GLANDS
GLOBINS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
LYSIS
MALES
MAMMALS
MAN
MATERIALS
METALS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PATHOLOGICAL CHANGES
PIGMENTS
PORPHYRINS
PRIMATES
PROTEINS
RODENTS
SYMPTOMS
TRANSITION ELEMENTS
VERTEBRATES