RESUMEN
We studied the effects of silver nanoparticles (10 nm) on HepaRG cell spheroids simulating liver tissue. The mathematical model was proposed that describes nanoparticle diffusion in a spheroid consisting of 5000 cells depending on the external nanoparticle concentration. It was demonstrated that cells in the 3D model were less sensitive to the toxic effects of nanoparticles in comparison with 2D cultures. Impaired integrity of the cell membrane did not deteriorate cell viability (according to MTT test).
Asunto(s)
Nanopartículas del Metal/toxicidad , Plata/toxicidad , Esferoides Celulares/fisiología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Hepatocitos/fisiología , Humanos , Plata/metabolismo , Esferoides Celulares/efectos de los fármacosRESUMEN
We created a physiologically substantiated kinetic model of cadmium transport and toxicity in intestinal cell model (Caco-2 cells). Transcriptome profiling of Caco-2 cells revealed high content of transporter DMT1 and ZIP14 and intensive expression of some calcium channels of the CACN family. The mathematical model describing three types of transporters, as well as intracellular cadmium binding with metallothionein and excretion through the basolateral membrane allowed us to construct cadmium uptake and transport curves that approximated the previously obtained experimental data. Using the proposed model, we determined toxic intracellular cadmium concentration leading to cell death and impairing the integrity of cell monolayer and described cadmium transport in this case.
Asunto(s)
Cadmio/metabolismo , Transporte Biológico , Células CACO-2 , Canales de Calcio/metabolismo , Proteínas de Transporte de Catión/metabolismo , Humanos , Cinética , Modelos BiológicosRESUMEN
СаÑо-2 cell line is widely used in in vitro studies of the intestinal wall permeability for drugs, but transport activity of these cells has not been thoroughly studied. We analyzed localization and expression of nucleoside transporters of the ENT family transferring a number of anticancer and antiviral drugs. Immunocytochemical staining revealed apical localization of ENT1 and integral expression of ENT2 in Caco-2 cells. Based on these data and on the value of hypoxanthine uptake by these cells, we created a mathematical model allowing quantification of transporter expression on the apical and basolateral membranes of Caco-2 cells. The discrepancy between gene and protein expression of the transporter complicating prediction of patient's sensitivity to the drug upon individual therapy selection is discussed.
Asunto(s)
Tranportador Equilibrativo 1 de Nucleósido/metabolismo , Transportador Equilibrativo 2 de Nucleósido/metabolismo , Mucosa Intestinal/metabolismo , Algoritmos , Células CACO-2 , Membrana Celular/metabolismo , Polaridad Celular , Tranportador Equilibrativo 1 de Nucleósido/genética , Transportador Equilibrativo 2 de Nucleósido/genética , Expresión Génica , Humanos , Modelos Biológicos , Transporte de ProteínasRESUMEN
Acetaminophen in a concentration of 5 mM increased the expression of JNK, HIF1A (hypoxiainduced factor), and CASP3, which indicated development of oxidative stress and apoptotic cell death. Acetaminophen in a concentration of 10 mM did not induce expression of HIF1A and CASP3, but reduced expression of chaperone HSP90, which attested to activation of a caspase-3-independent mechanism of cell death. The methods of preventing acetaminophen intoxication are discussed.
Asunto(s)
Acetaminofén/farmacología , Analgésicos no Narcóticos/farmacología , Caspasa 3/metabolismo , Neuronas/efectos de los fármacos , Proteína p53 Supresora de Tumor/metabolismo , Apoptosis , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Humanos , Estrés OxidativoRESUMEN
We studied regulation of hypoxanthine transport depending on its concentration in the culture medium. Caco-2 cells were differentiated on membrane filters to create a model of the intestine. Different hypoxanthine uptake on the apical and basolateral cell membranes was observed. The expression of SLC29 family genes encoding passive nucleoside transporters increased upon changes in hypoxanthine concentration in the medium Localization of the transporters and their influence on the effect of pharmacological preparations are discussed.
Asunto(s)
Colon/patología , Células Epiteliales/efectos de los fármacos , Tranportador Equilibrativo 1 de Nucleósido/fisiología , Transportador Equilibrativo 2 de Nucleósido/fisiología , Hipoxantina/farmacología , Purinas/metabolismo , Pirimidinas/metabolismo , Transporte Biológico , Línea Celular Tumoral , Neoplasias del Colon/metabolismo , Neoplasias del Colon/patología , Células Epiteliales/metabolismo , Tranportador Equilibrativo 1 de Nucleósido/biosíntesis , Tranportador Equilibrativo 1 de Nucleósido/genética , Transportador Equilibrativo 2 de Nucleósido/biosíntesis , Transportador Equilibrativo 2 de Nucleósido/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Familia de Multigenes , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/fisiologíaRESUMEN
Metabolome analysis of the serum from pregnant patients aimed at detection of low-molecular-weight biomarkers of gestation process disorders indicated a relationship between the metabolic profile of maternal serum and risk of gestosis. In women with pre-eclampsia or preterm delivery, analysis of serum purine metabolites revealed changes in the metabolite concentrations, associated with pregnancy complications.
Asunto(s)
Hipoxantina/sangre , Preeclampsia/sangre , Ácido Úrico/sangre , Xantina/sangre , Adulto , Biomarcadores/sangre , Cromatografía de Fase Inversa , Femenino , Edad Gestacional , Humanos , Recién Nacido , Recien Nacido Prematuro , Preeclampsia/diagnóstico , Preeclampsia/fisiopatología , Embarazo , RiesgoRESUMEN
The content of low-molecular-weight components in blood serum was studied by tandem mass-spectrometry in pregnant women. Serum metabolic profiles of patients with a grave obstetrical history were detected. The most significant changes were observed for the concentrations of low-molecular-weight substances involved in glucogenesis and ß-oxidation processes and in metabolic chains involving carbohydrates, carnitines, amino acids, and lipids.