ABSTRACT
The toxic effect of Ðß-oligomers accompanies chronic inflammation, with cytokines as main mediators. Therefore, the cytokine link of inflammation becomes a new target on the way to restrain amyloidosis. The aim of the study was the effect of aggregated Ðß42 on the dynamics of expression and formation of endogenous Ðß40 and cytokines (IL-1ß, TNFα, IL-6, IL-10) by peripheral blood mononuclear cells in vitro and its correction by curcumin. A suspension of mononuclear cells isolated ex tempore using ficoll-urografin gradient from venous blood samples of healthy volunteers were used to study the effects of Ðß42 (15 nM), curcumin (54 pM), and their combined action (at similar concentrations) in time dynamics: 0, 1, 3, 6 and 24 h incubation at 37 °C. Polymerase chain reaction with appropriate primers was used to determine the relative expression of mRNA for AßPP, TNFα, IL-1ß, IL-6, IL-10 and enzyme-linked immunosorbent assay to determine the content of Ðß40 and cytokines in mononuclear suspension during all periods of incubation. The individual dynamics AßPP and cytokine expression was shown under the action of the Aß42, which had influence on the content of Aß40, TNFα, IL-1ß, IL-6 and IL-10 in mononuclear suspension. Curcumin displayed the inhibitory effect on gene expression of AßPP, TNFα and IL6, which resulted in the decrease of the level of these two cytokines and Aß40. Thus, the dynamics of anti-inflammatory effect of curcumin in vitro for transcriptional and translational levels of cytokine's formation by mononuclear cells was shown in the work. Direct inhibitory effect of curcumin on the concentration of endogenous Aß40 during the 24 h incubation in conditions of toxic action of Aß42 aggregates was established.
Subject(s)
Amyloid beta-Peptides/genetics , Amyloid beta-Peptides/pharmacology , Amyloid beta-Protein Precursor/genetics , Interleukin-10/genetics , Interleukin-1beta/genetics , Interleukin-6/genetics , Peptide Fragments/genetics , Peptide Fragments/pharmacology , Tumor Necrosis Factor-alpha/genetics , Amyloid beta-Peptides/antagonists & inhibitors , Amyloid beta-Peptides/immunology , Amyloid beta-Protein Precursor/immunology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Curcumin/pharmacology , Gene Expression Regulation , Humans , Interleukin-10/immunology , Interleukin-1beta/immunology , Interleukin-6/immunology , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Peptide Fragments/antagonists & inhibitors , Peptide Fragments/immunology , Primary Cell Culture , Signal Transduction , Tumor Necrosis Factor-alpha/immunologyABSTRACT
OBJECTIVE: To estimate frequencies of polymorphic variants of TP53 codon 72 in the Ukrainian population. MATERIALS AND METHODS: We determined the allele frequencies for 148 healthy people. Genotyping was performed by allele specific polymerase chain reaction. RESULTS: We identified 31 individuals (20.9 %) with Arg/Arg genotype, Arg/Pro genotype was identified for 116 indi viduals (78.4 %), whereas genotype Pro/Pro was rare and was found in one person only (0.7 %). Genotype distribu tions were not within Hardy Weinberg equilibrium (χ2 = 59,7, p < 0.0001). CONCLUSIONS: Arg and Pro allele frequencies in the population of Ukraine are 60 and 40 % respectively, which is sig nificantly differ from the frequencies described in the literature for Poland, the Czech Republic, the USA and Brazil.
