ABSTRACT
We analyzed hepatotoxicity of three drugs: acetaminophen, metformin, and isoniazid. Spheroids of differentiated HepaRG cells cultured under microfluidic conditions were used as the model. Acute toxicity of substances was assessed by analyzing cell viability, while lactate concentration in the culture medium was used as the potential marker for evaluation of chronic exposure and non-lethal side effects of xenobiotics. The results were compared with mitochondrial activity and DNA fragmentation data. The efficiency and possibility of applying the integrated approach for assessment of drug hepatotoxicity are discussed.
Subject(s)
Hepatocytes/drug effects , Microfluidics/methods , Xenobiotics/toxicity , Cell Line , DNA Fragmentation/drug effects , Hepatocytes/metabolism , Humans , Toxicity Tests/methodsABSTRACT
Effects of cisplatin on the hepatic HepaRG cells cultured in spheroids were estimated using biochemical, cytofluorometric, and molecular methods. Hepatocyte viability and expression of mRNA of stress-dependent genes involved in the cell response to toxic agent cisplatin underwent time- and dose-dependent changes. Activation of oxidative stress was observed at the early stages of incubation (3 h) followed by induction of apoptosis after prolonging the incubation to 24 h. The prospects of using HepaRG cells cultured in spheroids for estimation of drug toxicity by variations in the expression of stress-dependent genes were demonstrated. An increase in expression of genes of GSR and HSPA1A proteins at the early stages of incubation with cisplatin can serve as a marker of the cytotoxic effects of cisplatin and other agents with similar mechanisms of action.
Subject(s)
Cisplatin/toxicity , Gene Expression , Hepatocytes/drug effects , Stress, Psychological , Base Sequence , Cell Line , Cytochrome P-450 CYP2E1/genetics , Cytochrome P-450 CYP3A/genetics , DNA Primers , Hepatocytes/enzymology , Hepatocytes/metabolism , Humans , Real-Time Polymerase Chain ReactionABSTRACT
We studied the possibility of using different methods for identification of mesenchymal multipotent stromal cells in the renal parenchyma under conditions of experimental thermal ischemia of the kidneys and acute pyelonephritis. In vivo and in vitro methods of identification of mesenchymal multipotent stromal cells by magnetic resonance imaging and immunological and immunohistochemical methods were compared. Labeling of stem cells with iron-containing particles followed by their histological identification and immunohistochemical staining with species-specific antibodies were the most informative methods. Active migration of the cells to the renal tissue was detected by these methods in experimental acute pyelonephritis with inflammation foci.