18 research outputs found
Protein carbonylation as a mechanism of regulation of MCF-7 breast cancer cell proliferation
Tumor progression is accompanied by dysregulation of cell proliferation and apoptosis, which plays an essential role in breast cancer pathogenesis and cell resistance to chemotherapy. The role of protein carbonylation in molecular mechanisms of regulating MCF-7 breast cancer cell proliferation under the effect of roscovitine, a selective inhibitor of cyclin-dependent kinases, was evaluated. The cells were grown in adherent cell culture with or without roscovitine. The levels of reduced/oxidized glutathione and the concentration of protein carbonyl derivatives were determined by spectrophotometry. The cell cycle was evaluated by the flow cytometry; the same technique was used to measure the reactive oxygen species (ROS) concentration. Cell culture with roscovitine resulted in a decrease in the redox potential of the glutathione system and a rise in the ROS and protein carbonyl derivative concentrations. Roscovitine contributed to the G0/G1 and G2/Π phase arrest due to its interaction with ATP-binding sites of cyclin-dependent kinases. Roscovitine could also promote enzyme carbonylation. The obtained results can be further used for development of personalized approaches to breast cancer therapy
ΠΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ Π±Π΅Π»ΠΊΠΎΠ² ΠΊΠ°ΠΊ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΡΠΉ ΡΠΏΠΎΡΠΎΠ± ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΠΈ ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ Π°Π΄Π΅Π½ΠΎΠΊΠ°ΡΡΠΈΠ½ΠΎΠΌΡ ΠΌΠΎΠ»ΠΎΡΠ½ΠΎΠΉ ΠΆΠ΅Π»Π΅Π·Ρ
Introduction. High rates of cancer incidence and mortality worldwide dictate the necessity of developing new methodological approaches in understanding the molecular mechanisms of cancer progression associated with intracellular redox regulation imbalance.The objective of the study was to evaluate the role of protein carbonylation in regulating breast cancer cell proliferation under redox status modulation.Materials and Methods. In the intact breast cancer cells and in the cells cultured under redox status modulation using 5mM N-ethylmaleimide (an - SH group blocker) and 5 Mm 1,4-dithioerythritol (a thiol group protector), the concentration of thioredoxin and its carbonylated form was measured using Western blot analysis. The activity of thioredoxin reductase and the level of protein carbonyl derivatives were determined using spectrophotometry. Cell cycle phase distribution was evaluated by flow cytometry.Results and Discussion. Under the effect of N-ethylmaleimide, cell cycle arrest in the S-phase was confirmed by oxidative modification of proteins, including thioredoxin carbonylation. When culturing MCF-7 cells in the presence of 1,4-dithioerythritol, cell cycle arrest in the G0/G1 phases was associated with a rise in the concentrations of reduced thioredoxin and glutathione forms.Conclusion. The thioredoxin system and oxidative modification of proteins are involved in redox-dependent modulation of breast cancer cell proliferation. Studies in the area of redox proteomics offer great potential to seek molecular targets of malignant transformation of breast cells.ΠΠ²Π΅Π΄Π΅Π½ΠΈΠ΅. ΠΠ΅ΡΠ΅ΡΠ΅Π½Π½Π°Ρ ΠΏΡΠΎΠ±Π»Π΅ΠΌΠ° ΡΠ°ΡΡΡΡΠ΅ΠΉ ΠΎΠ½ΠΊΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΉ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π΅ΠΌΠΎΡΡΠΈ ΠΈ ΡΠΌΠ΅ΡΡΠ½ΠΎΡΡΠΈ Π² ΠΌΠΈΡΠ΅ ΡΡΠ°Π²ΠΈΡ Π·Π°Π΄Π°ΡΡ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠΈ Π½ΠΎΠ²ΡΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΏΠΎΠ΄Ρ
ΠΎΠ΄ΠΎΠ² Π² ΠΏΠΎΠ½ΠΈΠΌΠ°Π½ΠΈΠΈ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΡ
ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠ³ΠΎ ΡΠΎΡΡΠ°, Π°ΡΡΠΎΡΠΈΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠ³ΠΎ Ρ Π΄ΠΈΡΠ±Π°Π»Π°Π½ΡΠΎΠΌ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ Π²Π½ΡΡΡΠΈΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ
ΡΠΈΡΡΠ΅ΠΌ.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ - ΡΡΡΠ°Π½ΠΎΠ²ΠΈΡΡ ΡΠΎΠ»Ρ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΠ΅Π΄ΠΎΠΊΡ-Π±Π΅Π»ΠΊΠΎΠ² Π² ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ Π°Π΄Π΅Π½ΠΎΠΊΠ°ΡΡΠΈΠ½ΠΎΠΌΡ ΠΌΠΎΠ»ΠΎΡΠ½ΠΎΠΉ ΠΆΠ΅Π»Π΅Π·Ρ ΠΏΡΠΈ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΠΈ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ°.ΠΠ°ΡΠ΅ΡΠΈΠ°Π» ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. Π ΠΈΠ½ΡΠ°ΠΊΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΊΠ°Ρ
Π°Π΄Π΅Π½ΠΎΠΊΠ°ΡΡΠΈΠ½ΠΎΠΌΡ ΠΌΠΎΠ»ΠΎΡΠ½ΠΎΠΉ ΠΆΠ΅Π»Π΅Π·Ρ ΠΈ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΡΠ΅ΠΌΡΡ
ΠΏΡΠΈ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΠΈ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ° Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ 5 ΠΌΠ N-ΡΡΠΈΠ»ΠΌΠ°Π»Π΅ΠΈΠΌΠΈΠ΄Π° (Π±Π»ΠΎΠΊΠ°ΡΠΎΡ SH-Π³ΡΡΠΏΠΏ Π±Π΅Π»ΠΊΠΎΠ² ΠΈ ΠΏΠ΅ΠΏΡΠΈΠ΄ΠΎΠ²) ΠΈ 5 ΠΌΠ 1,4-Π΄ΠΈΡΠΈΠΎΡΡΠΈΡΡΠΈΡΠΎΠ»Π° (ΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡ ΡΠΈΠΎΠ»ΠΎΠ²ΡΡ
Π³ΡΡΠΏΠΏ) ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ ΡΠΈΠΎΡΠ΅Π΄ΠΎΠΊΡΠΈΠ½Π° ΠΈ Π΅Π³ΠΎ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ ΡΠΎΡΠΌΡ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ Π²Π΅ΡΡΠ΅ΡΠ½-Π±Π»ΠΎΡΡΠΈΠ½Π³Π°; Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΡΠΈΠΎΡΠ΅Π΄ΠΎΠΊΡΠΈΠ½ΡΠ΅Π΄ΡΠΊΡΠ°Π·Ρ ΠΈ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΡ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΡΠ½ΡΡ
ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄Π½ΡΡ
Π±Π΅Π»ΠΊΠΎΠ² ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΡΠΏΠ΅ΠΊΡΡΠΎΡΠΎΡΠΎΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ; ΡΠ°ΡΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΠΎ ΡΠ°Π·Π°ΠΌ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ ΡΠΈΠΊΠ»Π° ΠΎΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΡΠ»ΡΠΎΡΠΈΠΌΠ΅ΡΡΠΈΠΈ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. ΠΡΠΈ Π΄Π΅ΠΉΡΡΠ²ΠΈΠΈ N-ΡΡΠΈΠ»ΠΌΠ°Π»Π΅ΠΈΠΌΠΈΠ΄Π° ΠΎΡΡΠ°Π½ΠΎΠ²ΠΊΠ° ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠΈ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π² S ΡΠ°Π·Π΅ Π±ΡΠ»Π° ΡΠ²ΡΠ·Π°Π½Π° c ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΠ΅ΠΉ Π±Π΅Π»ΠΊΠΎΠ², Π² ΡΠΎΠΌ ΡΠΈΡΠ»Π΅ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠΈΠΎΡΠ΅Π΄ΠΎΠΊΡΠΈΠ½Π°. ΠΡΡΠ°Π½ΠΎΠ²ΠΊΠ° ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ ΡΠΈΠΊΠ»Π° Π² G0/G1 ΡΠ°Π·Π°Ρ
ΠΏΡΠΈ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ ΠΠ‘F-7 Π² ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΠΈ 1,4-Π΄ΠΈΡΠΈΠΎΡΡΠΈΡΡΠΈΡΠΎΠ»Π° ΡΠΎΠΏΡΠΎΠ²ΠΎΠΆΠ΄Π°Π»Π°ΡΡ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠ΅ΠΌ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ Π²ΠΎΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½Π½ΡΡ
ΡΠΎΡΠΌ ΡΠΈΠΎΡΠ΅Π΄ΠΎΠΊΡΠΈΠ½Π° ΠΈ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π°.ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. Π Π΅Π΄ΠΎΠΊΡ-Π·Π°Π²ΠΈΡΠΈΠΌΠ°Ρ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΡ ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ Π°Π΄Π΅Π½ΠΎΠΊΠ°ΡΡΠΈΠ½ΠΎΠΌΡ ΠΌΠΎΠ»ΠΎΡΠ½ΠΎΠΉ ΠΆΠ΅Π»Π΅Π·Ρ ΠΎΡΡΡΠ΅ΡΡΠ²Π»ΡΠ΅ΡΡΡ ΠΏΡΠΈ ΡΡΠ°ΡΡΠΈΠΈ ΡΠΈΡΡΠ΅ΠΌΡ ΡΠΈΠΎΡΠ΅Π΄ΠΎΠΊΡΠΈΠ½Π° ΠΈ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π±Π΅Π»ΠΊΠΎΠ². ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ Π² ΠΎΠ±Π»Π°ΡΡΠΈ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΡΡΡΡ ΠΏΠ΅ΡΡΠΏΠ΅ΠΊΡΠΈΠ²Π½ΡΠΌΠΈ Π΄Π»Ρ ΠΏΠΎΠΈΡΠΊΠ° Π½ΠΎΠ²ΡΡ
ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΡ
ΠΌΠΈΡΠ΅Π½Π΅ΠΉ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠΉ ΡΡΠ°Π½ΡΡΠΎΡΠΌΠ°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΌΠΎΠ»ΠΎΡΠ½ΠΎΠΉ ΠΆΠ΅Π»Π΅Π·Ρ
ΠΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΠ΅ ΠΌΠ΅Ρ Π°Π½ΠΈΠ·ΠΌΡ Π²Π»ΠΈΡΠ½ΠΈΡ N-ΡΡΠΈΠ»ΠΌΠ°Π»Π΅ΠΈΠΌΠΈΠ΄Π° ΠΈ 1,4-Π΄ΠΈΡΠΈΠΎΡΡΠΈΡΡΠΈΡΠΎΠ»Π° Π½Π° ΡΠ΅Π³ΡΠ»ΡΡΠΈΡ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΎΠΏΡΡ ΠΎΠ»Π΅Π²ΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Π 19 ΠΏΡΠΈ Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈ
Relevance. Impairment of apoptosis regulation in P19 cells is correlated with generation of oxidative stress. UnderΒ hypoxia, changes in mitochondrial functions occur, which may exacerbate oxidative stress in the tumor cell.Β The aim of the study was to evaluate the effects of N-ethylmaleimide and 1,4-dithioerythritol on implementationΒ and regulation of apoptosis in P19 cells under hypoxia in vitro.Materials and methods. P19 cells (mouse teratocarcinoma) cultured under hypoxia served as the material for theΒ study. For redox status modulation, 5mM N-ethylmaleimide and 1,4-dithioerythritol in the final concentrations ofΒ 5 mM were used. The intracellular concentration of calcium ions, the transmembrane potential and the number ofΒ Annexin V, CD95 and CD120 positive cells were determined by flow cytometry. The levels of reduced, oxidizedΒ and protein-bound glutathione, protein SH groups, hydroxyl radical and protein carbonyl derivatives were measuredΒ by spectrophotometry.Results. The alteration in the redox status of the glutathione system under hypoxia, accompanied by oxidativeΒ modification of proteins (glutathionylation and carbonylation), influences the metabolism in the tumor cell onΒ the whole. Under the effects of 1,4-dithioerythritol, an SH group protector, this alteration promotes formation ofΒ additional mechanisms to escape apoptosis, whereas under the effects of N-ethylmaleimide, an SH group blocker,Β it, on the contrary, promotes apoptosis activation.Conclusions. The changes in the redox homeostasis of the tumor cell and modulation of oxidative modificationΒ of proteins (glutathionylation and carbonylation) under hypoxia are one of the promising approaches to targetedΒ regulation of cell death.ΠΠΊΡΡΠ°Π»ΡΠ½ΠΎΡΡΡ. ΠΠ°ΡΡΡΠ΅Π½ΠΈΠ΅ ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° Π² ΡΠΏΠΈΡΠ΅Π»ΠΈΠ°Π»ΡΠ½ΡΡ
ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΊΠ°Ρ
Π»ΠΈΠ½ΠΈΠΈ Π 19 ΡΠΎΠΏΡΡΠΆΠ΅Π½ΠΎΒ Ρ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΡΡΠ°. Π ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈ ΠΏΡΠΎΠΈΡΡ
ΠΎΠ΄ΠΈΡ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ ΡΡΠ½ΠΊΡΠΈΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΡΒ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΉ, ΡΡΠΎ ΠΌΠΎΠΆΠ΅Ρ Π²ΡΡΡΡΠΏΠ°ΡΡ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΡΠΌ ΡΠ°ΠΊΡΠΎΡΠΎΠΌ, ΡΡΡΠ³ΡΠ±Π»ΡΡΡΠΈΠΌΒ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΡΠΉ ΡΡΡΠ΅ΡΡ Π²Β ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠΉ ΠΊΠ»Π΅ΡΠΊΠ΅.Π¦Π΅Π»Ρ β ΠΎΡΠ΅Π½ΠΈΡΡ Π²Π»ΠΈΡΠ½ΠΈΠ΅ N-ΡΡΠΈΠ»ΠΌΠ°Π»Π΅ΠΈΠΌΠΈΠ΄Π° ΠΈ 1,4-Π΄ΠΈΡΠΈΠΎΡΡΠΈΡΡΠΈΡΠΎΠ»Π° Π½Π° ΡΠ΅Π°Π»ΠΈΠ·Π°ΡΠΈΡ ΠΈ ΡΠ΅Π³ΡΠ»ΡΡΠΈΡ Π°ΠΏΠΎΠΏΡΠΎΠ·Π°Β ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Π 19 ΠΏΡΠΈ Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈ in vitro.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»ΠΎΠΌ Π΄Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΡΠ»ΡΠΆΠΈΠ»ΠΈ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈΒ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΠ΅ ΠΊΠ»Π΅ΡΠΊΠΈ Π»ΠΈΠ½ΠΈΠΈ Π 19 (ΡΠ΅ΡΠ°ΡΠΎΠΊΠ°ΡΡΠΈΠ½ΠΎΠΌΠ° ΠΌΡΡΠΈ). ΠΠ»Ρ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΠΈ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈΒ N-ΡΡΠΈΠ»ΠΌΠ°Π»Π΅ΠΈΠΌΠΈΠ΄ Π² ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ 5 ΠΌΠ ΠΈ ΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡ SH-Π³ΡΡΠΏΠΏ β 1,4-Π΄ΠΈΡΠΈΠΎΡΡΠΈΡΡΠΈΡΠΎΠ» Π² ΠΊΠΎΠ½Π΅ΡΠ½ΠΎΠΉΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ 5 ΠΌΠ. ΠΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΡΠ»ΡΠΎΡΠΈΠΌΠ΅ΡΡΠΈΠΈ ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ Π²Π½ΡΡΡΠΈΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ΅ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅Β ΠΈΠΎΠ½ΠΎΠ² ΠΊΠ°Π»ΡΡΠΈΡ, ΡΡΠ°Π½ΡΠΌΠ΅ΠΌΠ±ΡΠ°Π½Π½ΡΠΉ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π» ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΉ, ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π°Π½Π½Π΅ΠΊΡΠΈΠ½ V-, CD95- ΠΈ CD120-ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ. ΠΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΡ Π²ΠΎΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½Π½ΠΎΠ³ΠΎ, ΠΎΠΊΠΈΡΠ»Π΅Π½Π½ΠΎΠ³ΠΎ ΠΈ Π±Π΅Π»ΠΊΠΎΠ²ΠΎ-ΡΠ²ΡΠ·Π°Π½Π½ΠΎΠ³ΠΎ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π°,Β SH-Π³ΡΡΠΏΠΏ ΠΏΡΠΎΡΠ΅ΠΈΠ½ΠΎΠ², Π³ΠΈΠ΄ΡΠΎΠΊΡΠΈΠ»ΡΠ½ΠΎΠ³ΠΎ ΡΠ°Π΄ΠΈΠΊΠ°Π»Π° ΠΈ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΡΠ½ΡΡ
ΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄Π½ΡΡ
Π±Π΅Π»ΠΊΠΎΠ² ΠΈΠ·ΠΌΠ΅ΡΡΠ»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌΒ ΡΠΏΠ΅ΠΊΡΡΠΎΡΠΎΡΠΎΠΌΠ΅ΡΠΈΠΈ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. Π ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ° ΡΠΈΡΡΠ΅ΠΌΡ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π°, ΡΠΎΠΏΡΠΎΠ²ΠΎΠΆΠ΄Π°ΡΡΠ΅Π΅ΡΡΒ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΠ΅ΠΉ Π±Π΅Π»ΠΊΠΎΠ² (Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΈ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅), ΠΎΠΊΠ°Π·ΡΠ²Π°Π΅Ρ Π²Π»ΠΈΡΠ½ΠΈΠ΅ Π½Π°Β ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΌ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠΉ ΠΊΠ»Π΅ΡΠΊΠΈ Π² ΡΠ΅Π»ΠΎΠΌ ΠΈ, ΠΏΡΠΈ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠΈ ΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠ° SH-Π³ΡΡΠΏΠΏ Π±Π΅Π»ΠΊΠΎΠ² β 1,4-Π΄ΠΈΡΠΈΠΎΡΡΠΈΡΡΠΈΡΠΎΠ»Π°, ΡΠΏΠΎΡΠΎΠ±ΡΡΠ²ΡΠ΅Ρ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΡΡΠΊΠΎΠ»ΡΠ·Π°Π½ΠΈΡ ΠΎΡ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π³ΠΈΠ±Π΅Π»ΠΈ, Π° Π²Β ΡΠ»ΡΡΠ°Π΅ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΡ Π±Π»ΠΎΠΊΠ°ΡΠΎΡΠ° SH-Π³ΡΡΠΏΠΏ ΠΏΡΠΎΡΠ΅ΠΈΠ½ΠΎΠ² β N-ΡΡΠΈΠ»ΠΌΠ°Π»Π΅ΠΈΠΌΠΈΠ΄Π° β Π°ΠΊΡΠΈΠ²Π°ΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π°.ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. Π ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ ΡΠ΅Π΄ΠΎΠΊΡ-Π³ΠΎΠΌΠ΅ΠΎΡΡΠ°Π·Π° ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠΉ ΠΊΠ»Π΅ΡΠΊΠΈ ΠΈ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΡΒ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π±Π΅Π»ΠΊΠΎΠ² (Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΈ ΠΊΠ°ΡΠ±ΠΎΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅) ΡΠ²Π»ΡΡΡΡΡ ΠΎΠ΄Π½ΠΈΠΌ ΠΈΠ·Β ΠΏΠ΅ΡΡΠΏΠ΅ΠΊΡΠΈΠ²Π½ΡΡ
ΠΏΠΎΠ΄Ρ
ΠΎΠ΄ΠΎΠ² ΡΠ°ΡΠ³Π΅ΡΠ½ΠΎΠΉ ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π³ΠΈΠ±Π΅Π»ΠΈ
Π£Π±ΠΈΠΊΠ²ΠΈΡΠΈΠ½ ΠΈ ΡΠ΅Π³ΡΠ»ΡΡΠΈΡ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΎΠΏΡΡ ΠΎΠ»Π΅Π²ΡΡ ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Jurkat
Introduction. One of the crucial tasks in medicine is studying the molecular mechanisms of selective management of tumor cell apoptosis following conformational changes in protein molecules (ubiquitination).The purpose of the study. The aim of the project is to establish the role of ubiquitin and ubiquitinligase in dexamethasone-induced apoptosis in Jurkat cells.Materials and methods. The study was carried out on the Jurkat tumor cell line (intact cells and cells cultured in the presence of an apoptosis inducer dexamethasone in the final concentration of 10 Β΅mol. In intact and dexamethasone-affected Jurkat cells, implementation of apoptosis and the amount of FAS-, TNF Receptor 1 and cells with reduced mitochondrial membrane potential were assessed by flow cytometry using FITC-conjugated Annexin V and Propidium Iodide. The levels of NF-ΞΊB, Apaf-1, ubiquitin and ubiquitin ligase were determined by Western blot analysis. The activity of caspase-3 was measured by spectrofluorometry.Results. When adding the apoptosis inducer dexamethasone to the Jurkat cell culture, we registered a fall in the concentration of ubiquitin and a rise in the level of ubiquitinligase against the backdrop of activated receptor(an increase in the amount of Annexin V positive cells, FASand TNF Receptor 1) and mitochondrialmediated (an increase in the number of cells with reduced mitochondrial membrane potential and elevation of Apaf-1 level) pathways of apoptosis, as opposed to the intact cell culture. We estimated the completion of apoptosis by determining the activity of caspase-3 in the investigated tumor cells.Conclusion. The obtained findings allow the conclusion that ubiquitination of regulatory and effector proteins in programmed cell death is one of the molecular mechanisms that regulates and selectively controls apoptosis in Jurkat cells.ΠΠΊΡΡΠ°Π»ΡΠ½ΠΎΡΡΡ. ΠΠ΄Π½ΠΎΠΉ ΠΈΠ· Π°ΠΊΡΡΠ°Π»ΡΠ½ΡΡ
Π·Π°Π΄Π°Ρ ΠΌΠ΅Π΄ΠΈΡΠΈΠ½Ρ ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΡ
ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΡΠ΅Π»Π΅ΠΊΡΠΈΠ²Π½ΠΎΠ³ΠΎ ΡΠΏΡΠ°Π²Π»Π΅Π½ΠΈΡ Π°ΠΏΠΎΠΏΡΠΎΡΠΈΡΠ΅ΡΠΊΠΎΠΉ Π³ΠΈΠ±Π΅Π»ΡΡ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π² ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠ΅ ΠΊΠΎΠ½ΡΠΎΡΠΌΠ°ΡΠΈΠΎΠ½Π½ΡΡ
ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠΉ Π±Π΅Π»ΠΊΠΎΠ²ΡΡ
ΠΌΠΎΠ»Π΅ΠΊΡΠ» (ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ). Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ: ΡΡΡΠ°Π½ΠΎΠ²ΠΈΡΡ ΡΠΎΠ»Ρ ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½Π° ΠΈ ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½Π»ΠΈΠ³Π°Π·Ρ Π² Π΄Π΅ΠΊΡΠ°ΠΌΠ΅ΡΠ°Π·ΠΎΠ½-ΠΈΠ½Π΄ΡΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΌ Π°ΠΏΠΎΠΏΡΠΎΠ·Π΅ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Jurkat.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΠ½ΡΠ°ΠΊΡΠ½ΡΠ΅ ΠΈ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½Π½ΡΠ΅ ΠΏΡΠΈ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΠΎΠΌ Π΄ΠΎΠ±Π°Π²Π»Π΅Π½ΠΈΠΈ ΠΈΠ½Π΄ΡΠΊΡΠΎΡΠ° Π°ΠΏΠΎΠΏΡΠΎΠ·Π° Π΄Π΅ΠΊΡΠ°ΠΌΠ΅ΡΠ°Π·ΠΎΠ½Π° Π² ΠΊΠΎΠ½Π΅ΡΠ½ΠΎΠΉ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ 10 ΠΌΠΊΠ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΠ΅ ΠΊΠ»Π΅ΡΠΊΠΈ Π»ΠΈΠ½ΠΈΠΈ Jurkat. ΠΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΡΠ»ΡΠΎΡΠΈΠΌΠ΅ΡΡΠΈΠΈ Π² ΠΈΠ½ΡΠ°ΠΊΡΠ½ΡΡ
ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΊΠ°Ρ
Π»ΠΈΠ½ΠΈΠΈ Jurkat ΠΈ ΠΏΠΎΡΠ»Π΅ ΠΏΡΠ΅Π΄Π²Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΡ Π΄Π΅ΠΊΡΠ°ΠΌΠ΅ΡΠ°Π·ΠΎΠ½ΠΎΠΌ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ ΠΎΡΠ΅Π½ΠΊΡ ΡΠ΅Π°Π»ΠΈΠ·Π°ΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ FITΠ‘-ΠΌΠ΅ΡΠ΅Π½Π½ΠΎΠ³ΠΎ Π°Π½Π½Π΅ΠΊΡΠΈΠ½Π° V ΠΈ ΠΏΡΠΎΠΏΠΈΠ΄ΠΈΡ ΠΈΠΎΠ΄ΠΈΠ΄Π°, ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π° FASΠΈ TNF-ΡΠ΅ΡΠ΅ΠΏΡΠΎΡ 1 ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΡΠΎ ΡΠ½ΠΈΠΆΠ΅Π½Π½ΡΠΌ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΠΌ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΠΎΠΌ. ΠΠ΅ΡΠΎΠ΄ΠΎΠΌ Π²Π΅ΡΡΠ΅ΡΠ½-Π±Π»ΠΎΡΡΠΈΠ½Π³Π° ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠΎΠ½Π½ΡΡ
ΡΠ°ΠΊΡΠΎΡΠΎΠ² NF-ΞΊB, Apaf-1; ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½Π° ΠΈ ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½Π»ΠΈΠ³Π°Π·Ρ; Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ°ΡΠΏΠ°Π·Ρ-3 ΡΠ΅Π³ΠΈΡΡΡΠΈΡΠΎΠ²Π°Π»ΠΈ ΡΠΏΠ΅ΠΊΡΡΠΎΡΠ»ΡΠΎΡΠΈΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. ΠΡΠΈ Π΄ΠΎΠ±Π°Π²Π»Π΅Π½ΠΈΠΈ Π΄Π΅ΠΊΡΠ°ΠΌΠ΅ΡΠ°Π·ΠΎΠ½Π° β ΠΈΠ½Π΄ΡΠΊΡΠΎΡΠ° Π°ΠΏΠΎΠΏΡΠΎΠ·Π° β Π² ΡΡΠ΅Π΄Ρ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Jurkat ΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½ΠΎ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½Π° ΠΈ Π²ΠΎΠ·ΡΠ°ΡΡΠ°Π½ΠΈΠ΅ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½Π»ΠΈΠ³Π°Π·Ρ Π½Π° ΡΠΎΠ½Π΅ Π°ΠΊΡΠΈΠ²Π°ΡΠΈΠΈ ΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠ½ΠΎΠ³ΠΎ (ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΠ΅ Π΄ΠΎΠ»ΠΈ Π°Π½Π½Π΅ΠΊΡΠΈΠ½-, FASΠΈ TNFΡΠ΅ΡΠ΅ΠΏΡΠΎΡ 1 ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ) ΠΈ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ (Π²ΠΎΠ·ΡΠ°ΡΡΠ°Π½ΠΈΠ΅ ΡΠΈΡΠ»Π° ΠΊΠ»Π΅ΡΠΎΠΊ ΡΠΎ ΡΠ½ΠΈΠΆΠ΅Π½Π½ΡΠΌ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΠΌ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΠΎΠΌ ΠΈ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΈΠΎΠ½Π½ΠΎΠ³ΠΎ ΡΠ°ΠΊΡΠΎΡΠ° Apaf-1) ΠΏΡΡΠ΅ΠΉ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΏΠΎ ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ Ρ ΠΈΠ½ΡΠ°ΠΊΡΠ½ΠΎΠΉ ΠΊΡΠ»ΡΡΡΡΠΎΠΉ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ. ΠΠ°Π²Π΅ΡΡΠ΅Π½Π½ΠΎΡΡΡ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΎΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ, ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ°ΡΠΏΠ°Π·Ρ-3 Π² ΠΈΠ·ΡΡΠ°Π΅ΠΌΡΡ
ΠΊΠ»Π΅ΡΠΊΠ°Ρ
.ΠΡΠ²ΠΎΠ΄Ρ. ΠΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΏΠΎΠ·Π²ΠΎΠ»ΡΡΡ ΡΠ΄Π΅Π»Π°ΡΡ Π²ΡΠ²ΠΎΠ΄ ΠΎ ΡΠΎΠΌ, ΡΡΠΎ ΠΏΡΠΎΡΠ΅ΡΡ ΡΠ±ΠΈΠΊΠ²ΠΈΡΠΈΠ½ΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π±Π΅Π»ΠΊΠΎΠ²-ΡΠ΅Π³ΡΠ»ΡΡΠΎΡΠΎΠ² ΠΈ Π±Π΅Π»ΠΊΠΎΠ²-ΡΡΡΠ΅ΠΊΡΠΎΡΠΎΠ² ΠΏΡΠΎΠ³ΡΠ°ΠΌΠΌΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ Π³ΠΈΠ±Π΅Π»ΠΈ ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΎΠ΄Π½ΠΈΠΌ ΠΈΠ· ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΡ
ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Jurkat
ΠΠΠ₯ΠΠΠΠΠΠ« ΠΠΠΠ ΠΠΠ£ΠΠ―Π¦ΠΠ ΠΠΠΠΠ’ΠΠΠ ΠΠΠ£Π₯ΠΠΠΠΠ«Π₯ ΠΠΠΠ’ΠΠ ΠΠΠΠΠ Π 19 Π Π£Π‘ΠΠΠΠΠ―Π₯ ΠΠΠΠ£ΠΠ―Π¦ΠΠ Π ΠΠΠΠΠ‘-Π‘Π’ΠΠ’Π£Π‘Π
Introduction. Changes in the redox status of tumor cells can be used as one of the molecular mechanisms of apoptosis aimed at increasing the susceptibility of tumor cells to chemotherapeutic agents. Purpose: to study the mechanisms of dysregulation of apoptosis in P19 tumor cells under the conditions of redox status modulation. Material and methods. Apoptosis in P19 tumor cells was assessed by flow cytometry analysis. The number of annexin-positive cells, the expression of CD95 and CD120, as well as the intracellular calcium ion concentration and the percentage of cells with reduced mitochondrial transmembrane potential were measured. The protein-glutathione mixed-disulfide level and the GSH/GSSG ratio were determined by spectrophotometry. To modulate redox status of cells, the protector and blocker of SH-groups, or N-acetylcysteine were used. Results. Incubation of cultures in the presence of SH-group blocker resulted in the imbalance in the glutathione system with increased concentration of glutathionylated proteins. A decreased redox status led to an increased CD95 and CD120 expression levels on the membrane of P19 tumor cells, as well as to decreased mitochondrial potential and increased intracellular calcium ion concentration, thus contributing to the launch of a P19 tumor cells. The presence of SH-group blocker and N-acetylcysteine resulted in an increased number of annexinpositive cells. Conclusion. Along with the development of oxidative stress, the molecular redox-dependent mechanisms of apoptosis dysregulation through the mitochondrial and receptor-mediated pathways were identified in the P19 tumor cells.ΠΠ²Π΅Π΄Π΅Π½ΠΈΠ΅. ΠΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ° ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΠΌΠΎΠΆΠ΅Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°ΡΡΡΡ ΠΊΠ°ΠΊ ΠΎΠ΄ΠΈΠ½ ΠΈΠ· ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΡ
ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π°, Π½Π°ΡΠ΅Π»Π΅Π½Π½ΡΠΉ Π½Π° ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΠ΅ Π²ΠΎΡΠΏΡΠΈΠΈΠΌΡΠΈΠ²ΠΎΡΡΠΈ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΠΊ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ Ρ
ΠΈΠΌΠΈΠΎΡΠ΅ΡΠ°ΠΏΠ΅Π²ΡΠΈΡΠ΅ΡΠΊΠΈΡ
Π°Π³Π΅Π½ΡΠΎΠ². Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ β ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΠΎΠ² Π΄ΠΈΠ·ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Π 19 Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ
ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΠΈ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ°. ΠΌΠ°ΡΠ΅ΡΠΈΠ°Π» ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. Π Ρ
ΠΎΠ΄Π΅ ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½Π½ΠΎΠ³ΠΎ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ½ΡΡ ΠΎΡΠ΅Π½ΠΊΡ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠΉ Π»ΠΈΠ½ΠΈΠΈ Π 19 ΠΎΡΡΡΠ΅ΡΡΠ²Π»ΡΠ»ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΡΠ»ΡΠΎΡΠΈΠΌΠ΅ΡΡΠΈΠΈ. ΠΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π°Π½Π½Π΅ΠΊΡΠΈΠ½ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ, ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ CD95 ΠΈ CD120, Π° ΡΠ°ΠΊΠΆΠ΅ ΠΏΡΠΎΡΠ΅Π½Ρ ΠΊΠ»Π΅ΡΠΎΠΊ ΡΠΎ ΡΠ½ΠΈΠΆΠ΅Π½Π½ΡΠΌ ΡΡΠ°Π½ΡΠΌΠ΅ΠΌΠ±ΡΠ°Π½Π½ΡΠΌ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΠΎΠΌ ΠΈ Π²Π½ΡΡΡΠΈΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΡ ΠΈΠΎΠ½ΠΎΠ² ΠΊΠ°Π»ΡΡΠΈΡ. Π‘ΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ Π±Π΅Π»ΠΊΠΎΠ²ΠΎΡΠ²ΡΠ·Π°Π½Π½ΠΎΠ³ΠΎ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° ΠΈ Π²Π΅Π»ΠΈΡΠΈΠ½Ρ ΡΠΎΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΡ Π²ΠΎΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½Π½ΠΎΠΉ ΡΠΎΡΠΌΡ ΡΡΠΈΠΏΠ΅ΠΏΡΠΈΠ΄Π° ΠΊ ΠΎΠΊΠΈΡΠ»Π΅Π½Π½ΠΎΠΉ ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΡΠΏΠ΅ΠΊΡΡΠΎΡΠΎΡΠΎΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ. ΠΠ»Ρ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΈΠΈ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ Π±Π»ΠΎΠΊΠ°ΡΠΎΡ ΠΈΠ»ΠΈ ΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡ SH-Π³ΡΡΠΏΠΏ, Π»ΠΈΠ±ΠΎ N-Π°ΡΠ΅ΡΠΈΠ»ΡΠΈΡΡΠ΅ΠΈΠ½. ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ. ΠΠ½ΠΊΡΠ±Π°ΡΠΈΡ ΠΊΡΠ»ΡΡΡΡΡ Π² ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΠΈ Π±Π»ΠΎΠΊΠ°ΡΠΎΡΠ° SH-Π³ΡΡΠΏΠΏ ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΠ»Π° ΠΊ Π΄ΠΈΡΠ±Π°Π»Π°Π½ΡΡ ΡΠΈΡΡΠ΅ΠΌΡ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° Π½Π° ΡΠΎΠ½Π΅ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΡ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ Π΅Π³ΠΎ ΡΡΠ°ΠΊΡΠΈΠΈ, ΡΠ²ΡΠ·Π°Π½Π½ΠΎΠΉ Ρ Π±Π΅Π»ΠΊΠ°ΠΌΠΈ. Π‘Π½ΠΈΠΆΠ΅Π½ΠΈΠ΅ ΡΠ΅Π΄ΠΎΠΊΡ-ΡΡΠ°ΡΡΡΠ° ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΠ»ΠΎ ΠΊ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΡ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ CD95 ΠΈ CD120 Π½Π° ΠΌΠ΅ΠΌΠ±ΡΠ°Π½Π΅ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΈ Π 19, Π° ΡΠ°ΠΊΠΆΠ΅ ΠΊ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΡ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»Π° ΠΈ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ Π²Π½ΡΡΡΠΈΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ ΠΈΠΎΠ½ΠΎΠ² ΠΊΠ°Π»ΡΡΠΈΡ, ΡΡΠΎ ΡΠΏΠΎΡΠΎΠ±ΡΡΠ²ΠΎΠ²Π°Π»ΠΎ Π·Π°ΠΏΡΡΠΊΡ Π°ΠΏΠΎΠΏΡΠΎΠ·Π°. ΠΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π°Π½Π½Π΅ΠΊΡΠΈΠ½-ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΡΠ²Π΅Π»ΠΈΡΠΈΠ²Π°Π»ΠΎΡΡ ΠΏΡΠΈ Π΄Π΅ΠΉΡΡΠ²ΠΈΠΈ Π±Π»ΠΎΠΊΠ°ΡΠΎΡΠ° SH-Π³ΡΡΠΏΠΏ ΠΈ Π² ΠΏΡΠΈΡΡΡΡΡΠ²ΠΈΠΈ N-Π°ΡΠ΅ΡΠΈΠ»ΡΠΈΡΡΠ΅ΠΈΠ½Π°. ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. Π ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΊΠ°Ρ
Π»ΠΈΠ½ΠΈΠΈ Π 19 Π½Π° ΡΠΎΠ½Π΅ ΡΠ°Π·Π²ΠΈΡΠΈΡ ΠΎΠΊΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΡΡΠ° Π²ΡΡΠ²Π»Π΅Π½Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΡΠ΅ ΡΠ΅Π΄ΠΎΠΊΡ-Π·Π°Π²ΠΈΡΠΈΠΌΡΠ΅ ΠΌΠ΅Ρ
Π°Π½ΠΈΠ·ΠΌΡ Π΄ΠΈΠ·ΡΠ΅Π³ΡΠ»ΡΡΠΈΠΈ Π°ΠΏΠΎΠΏΡΠΎΠ·Π° ΠΏΠΎ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΠΎΠΌΡ ΠΈ ΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠΏΠΎΡΡΠ΅Π΄ΠΎΠ²Π°Π½Π½ΠΎΠΌΡ ΠΏΡΡΠΈ
ΠΠ°ΡΠΈΠ°Π½ΡΡ ΡΠ΅ΡΠ΅Π½ΠΈΡ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° Ρ Π΄Π΅ΡΠ΅ΠΉ ΠΈ Π²ΠΊΠ»Π°Π΄ Π°ΠΊΡΠΈΠ²Π°ΡΠΈΠΈ ΠΏΠ΅ΡΠ΅ΠΊΠΈΡΠ½ΠΎΠ³ΠΎ ΠΎΠΊΠΈΡΠ»Π΅Π½ΠΈΡ Π»ΠΈΠΏΠΈΠ΄ΠΎΠ², Π΄ΠΈΡΠ±Π°Π»Π°Π½ΡΠ° Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠ½ΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΡ Π² ΠΈΡ ΡΠ°Π·Π²ΠΈΡΠΈΠ΅
The objective is to compare clinical symptoms, lipid peroxidation indicators, the state of the antioxidant system and assess their impact on the severity and progression of pseudotuberculosis in children.Materials and methods. We examined 125 children with pseudotuberculosis divided into 4 groups according to the severity and nature of the disease progression and 45 healthy children. The material for the study was red blood cells and blood plasma of patients received in the dynamics β the acute period (during hospitalization); 3-4 weeks later β the phase of early convalescence with a non-smooth progression and moderate and heavy severity; the recovery period with a smooth progression and mild and moderate severity; 5-6 weeks later β the recovery period with a non-smooth progression and moderate and heavy severity. The spectrophotometric method was used to study lipid peroxidation (the concentration of diene conjugates, TBA-reactive substances) in the blood plasma and components of the antioxidant support system (the content of reduced glutathione; the activity of glutathione reductase, glutathione peroxidase, and catalase) in red blood cells.Results. It was determined that moderate and heavy pseudotuberculosis forms prevail in hospitalized children, the disease progression in 35.2% of them was non-smooth; lipid peroxidation products accumulate in the blood plasma and the concentration of reduced glutathione decreases in red blood cells during the acute period of pseudotuberculosis in all children relative to the parameters in the control group. In the period of early convalescence an imbalance in the functioning of antioxidant enzymes of red blood cells, as well as the accumulation of TBA-reactive substances and a decrease in the content of reduced glutathione were observed in patients with moderate and heavy pseudotuberculosis.Conclusion. The impact of the imbalance of pro-/antioxidants on the formation of predominantly moderate and heavy pseudotuberculosis in children is shown. Prognostic criteria for the development of a non-smooth progression of pseudotuberculosis are a high level of lipid peroxidation products in the blood plasma, no normalization in values of glutathioneΒ system components and the activity of erythrocyte catalase during early convalescence.Β Π¦Π΅Π»Ρ: ΡΠΎΠΏΠΎΡΡΠ°Π²ΠΈΡΡ ΠΊΠ»ΠΈΠ½ΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΡΠΈΠΌΠΏΡΠΎΠΌΡ, ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»ΠΈ ΠΏΠ΅ΡΠ΅ΠΊΠΈΡΠ½ΠΎΠ³ΠΎ ΠΎΠΊΠΈΡΠ»Π΅Π½ΠΈΡ Π»ΠΈΠΏΠΈΠ΄ΠΎΠ², ΡΠΎΡΡΠΎΡΠ½ΠΈΡ Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠ½ΠΎΠΉ ΡΠΈΡΡΠ΅ΠΌΡ ΠΈ ΠΎΡΠ΅Π½ΠΈΡΡ ΠΈΡ
Π²ΠΊΠ»Π°Π΄ Π² ΡΡΠ΅ΠΏΠ΅Π½Ρ ΡΡΠΆΠ΅ΡΡΠΈ ΠΈ Π²Π°ΡΠΈΠ°Π½Ρ ΡΠ΅ΡΠ΅Π½ΠΈΡ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° Ρ Π΄Π΅ΡΠ΅ΠΉ.ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΠ±ΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΎ 125 Π΄Π΅ΡΠ΅ΠΉ, Π±ΠΎΠ»ΡΠ½ΡΡ
ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·ΠΎΠΌ, ΡΠ°Π·Π΄Π΅Π»Π΅Π½Π½ΡΡ
Π½Π° 4 Π³ΡΡΠΏΠΏΡ ΠΏΠΎ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΡΡΠΆΠ΅ΡΡΠΈ ΠΈ Ρ
Π°ΡΠ°ΠΊΡΠ΅ΡΡ ΡΠ΅ΡΠ΅Π½ΠΈΡ Π±ΠΎΠ»Π΅Π·Π½ΠΈ, ΠΈ 45 Π·Π΄ΠΎΡΠΎΠ²ΡΡ
Π΄Π΅ΡΠ΅ΠΉ. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»ΠΎΠΌ Π΄Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΡΠ»ΡΠΆΠΈΠ»ΠΈ ΡΡΠΈΡΡΠΎΡΠΈΡΡ ΠΈ ΠΏΠ»Π°Π·ΠΌΠ° ΠΊΡΠΎΠ²ΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ², ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ Π² Π΄ΠΈΠ½Π°ΠΌΠΈΠΊΠ΅ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΡ β ΠΎΡΡΡΡΠΉ ΠΏΠ΅ΡΠΈΠΎΠ΄ (ΠΏΡΠΈ Π³ΠΎΡΠΏΠΈΡΠ°Π»ΠΈΠ·Π°ΡΠΈΠΈ); ΡΠ΅ΡΠ΅Π· 3β4 Π½Π΅Π΄Π΅Π»ΠΈ β ΡΠ°Π·Π° ΡΠ°Π½Π½Π΅ΠΉ ΡΠ΅ΠΊΠΎΠ½Π²Π°Π»Π΅ΡΡΠ΅Π½ΡΠΈΠΈ ΠΏΡΠΈ Π½Π΅Π³Π»Π°Π΄ΠΊΠΎΠΌ ΡΠ΅ΡΠ΅Π½ΠΈΠΈ ΡΡΠ΅Π΄Π½Π΅ΠΉ ΠΈ ΡΡΠΆΠ΅Π»ΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΡΡΠΆΠ΅ΡΡΠΈ; ΠΏΠ΅ΡΠΈΠΎΠ΄ Π²ΡΠ·Π΄ΠΎΡΠΎΠ²Π»Π΅Π½ΠΈΡ ΠΏΡΠΈ Π³Π»Π°Π΄ΠΊΠΎΠΌ ΡΠ΅ΡΠ΅Π½ΠΈΠΈ Π»Π΅Π³ΠΊΠΎΠΉ ΠΈ ΡΡΠ΅Π΄Π½Π΅ΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΡΡΠΆΠ΅ΡΡΠΈ; ΡΠ΅ΡΠ΅Π· 5β6 Π½Π΅Π΄Π΅Π»Ρ β ΠΏΠ΅ΡΠΈΠΎΠ΄ Π²ΡΠ·Π΄ΠΎΡΠΎΠ²Π»Π΅Π½ΠΈΡ ΠΏΡΠΈ Π½Π΅Π³Π»Π°Π΄ΠΊΠΎΠΌ ΡΠ΅ΡΠ΅Π½ΠΈΠΈ ΡΡΠ΅Π΄Π½Π΅ΠΉ ΠΈ ΡΡΠΆΠ΅Π»ΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΡΡΠΆΠ΅ΡΡΠΈ. Π‘ΠΏΠ΅ΠΊΡΡΠΎΡΠΎΡΠΎΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π»ΠΈ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»ΠΈ ΠΏΠ΅ΡΠ΅ΠΊΠΈΡΠ½ΠΎΠ³ΠΎ ΠΎΠΊΠΈΡΠ»Π΅Π½ΠΈΡ Π»ΠΈΠΏΠΈΠ΄ΠΎΠ² (ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΡ Π΄ΠΈΠ΅Π½ΠΎΠ²ΡΡ
ΠΊΠΎΠ½ΡΡΠ³Π°ΡΠΎΠ², Π’ΠΠ-ΡΠ΅Π°ΠΊΡΠΈΠ²Π½ΡΡ
ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ²) Π² ΠΏΠ»Π°Π·ΠΌΠ΅ ΠΊΡΠΎΠ²ΠΈ ΠΈ ΠΊΠΎΠΌΠΏΠΎΠ½Π΅Π½ΡΠΎΠ² ΡΠΈΡΡΠ΅ΠΌΡ Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠ½ΠΎΠΉ Π·Π°ΡΠΈΡΡ (ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΠ΅ Π²ΠΎΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½Π½ΠΎΠ³ΠΎ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π°; Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½ΡΠ΅Π΄ΡΠΊΡΠ°Π·Ρ, Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½ΠΏΠ΅ΡΠΎΠΊΡΠΈΠ΄Π°Π·Ρ, ΠΊΠ°ΡΠ°Π»Π°Π·Ρ) Π² ΡΡΠΈΡΡΠΎΡΠΈΡΠ°Ρ
.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. Π£ΡΡΠ°Π½ΠΎΠ²ΠΈΠ»ΠΈ ΠΏΡΠ΅ΠΎΠ±Π»Π°Π΄Π°Π½ΠΈΠ΅ ΡΡΠ΅Π΄Π½Π΅ΠΉ ΠΈ ΡΡΠΆΠ΅Π»ΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° Ρ Π³ΠΎΡΠΏΠΈΡΠ°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Π΄Π΅ΡΠ΅ΠΉ, Π½Π΅Π³Π»Π°Π΄ΠΊΠΎΠ΅ ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ Ρ 35,2% ΠΈΠ· Π½ΠΈΡ
; Π½Π°ΠΊΠΎΠΏΠ»Π΅Π½ΠΈΠ΅ ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² ΠΏΠ΅ΡΠ΅ΠΊΠΈΡΠ½ΠΎΠ³ΠΎ ΠΎΠΊΠΈΡΠ»Π΅Π½ΠΈΡ Π»ΠΈΠΏΠΈΠ΄ΠΎΠ² Π² ΠΏΠ»Π°Π·ΠΌΠ΅ ΠΊΡΠΎΠ²ΠΈ ΠΈ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ Π²ΠΎΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½Π½ΠΎΠ³ΠΎ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° Π² ΡΡΠΈΡΡΠΎΡΠΈΡΠ°Ρ
Π² ΠΎΡΡΡΡΠΉ ΠΏΠ΅ΡΠΈΠΎΠ΄ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° Ρ Π²ΡΠ΅Ρ
Π΄Π΅ΡΠ΅ΠΉ ΠΎΡΠ½ΠΎΡΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ΠΉ Π² Π³ΡΡΠΏΠΏΠ΅ ΠΊΠΎΠ½ΡΡΠΎΠ»Ρ. Π ΠΏΠ΅ΡΠΈΠΎΠ΄ ΡΠ°Π½Π½Π΅ΠΉ ΡΠ΅ΠΊΠΎΠ½Π²Π°Π»Π΅ΡΡΠ΅Π½ΡΠΈΠΈ Ρ Π±ΠΎΠ»ΡΠ½ΡΡ
ΡΡΠ΅Π΄Π½Π΅ΠΉ ΠΈ ΡΡΠΆΠ΅Π»ΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΡΡΠΆΠ΅ΡΡΠΈ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° ΠΎΠ΄Π½ΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΠΎ Ρ Π½Π°ΠΊΠΎΠΏΠ»Π΅Π½ΠΈΠ΅ΠΌ Π’ΠΠ-ΡΠ΅Π°ΠΊΡΠΈΠ²Π½ΡΡ
ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² ΠΈ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ΠΌ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ Π²ΠΎΡΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½Π½ΠΎΠ³ΠΎ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° Π½Π°Π±Π»ΡΠ΄Π°Π»ΡΡ Π΄ΠΈΡΠ±Π°Π»Π°Π½Ρ Π² ΡΠ°Π±ΠΎΡΠ΅ ΡΠ΅ΡΠΌΠ΅Π½ΡΠΎΠ² Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠ½ΠΎΠΉ Π·Π°ΡΠΈΡΡ ΡΡΠΈΡΡΠΎΡΠΈΡΠΎΠ².ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. ΠΠΎΠΊΠ°Π·Π°Π½ Π²ΠΊΠ»Π°Π΄ Π΄ΠΈΡΠ±Π°Π»Π°Π½ΡΠ° ΠΏΡΠΎ-/Π°Π½ΡΠΈΠΎΠΊΡΠΈΠ΄Π°Π½ΡΠΎΠ² Π² ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠ΅ΠΈΠΌΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎ ΡΡΠ΅Π΄Π½Π΅ΠΉ ΠΈ ΡΡΠΆΠ΅Π»ΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½Π΅ΠΉ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° Ρ Π΄Π΅ΡΠ΅ΠΉ. ΠΡΠΎΠ³Π½ΠΎΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌΠΈ ΠΊΡΠΈΡΠ΅ΡΠΈΡΠΌΠΈ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π½Π΅Π³Π»Π°Π΄ΠΊΠΎΠ³ΠΎ ΡΠ΅ΡΠ΅Π½ΠΈΡ ΠΏΡΠ΅Π²Π΄ΠΎΡΡΠ±Π΅ΡΠΊΡΠ»Π΅Π·Π° ΡΠ²Π»ΡΡΡΡΡ Π²ΡΡΠΎΠΊΠΈΠΉ ΡΡΠΎΠ²Π΅Π½Ρ ΠΏΡΠΎΠ΄ΡΠΊΡΠΎΠ² ΠΏΠ΅ΡΠ΅ΠΊΠΈΡΠ½ΠΎΠ³ΠΎ ΠΎΠΊΠΈΡΠ»Π΅Π½ΠΈΡ Π»ΠΈΠΏΠΈΠ΄ΠΎΠ² Π² ΠΏΠ»Π°Π·ΠΌΠ΅ ΠΊΡΠΎΠ²ΠΈ, ΠΎΡΡΡΡΡΡΠ²ΠΈΠ΅ Π½ΠΎΡΠΌΠ°Π»ΠΈΠ·Π°ΡΠΈΠΈ ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ΠΉ ΠΊΠΎΠΌΠΏΠΎΠ½Π΅Π½ΡΠΎΠ² ΡΠΈΡΡΠ΅ΠΌΡ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° ΠΈ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΊΠ°ΡΠ°Π»Π°Π·Ρ ΡΡΠΈΡΡΠΎΡΠΈΡΠΎΠ² Π² ΠΏΠ΅ΡΠΈΠΎΠ΄ ΡΠ°Π½Π½Π΅ΠΉ ΡΠ΅ΠΊΠΎΠ½Π²Π°Π»Π΅ΡΡΠ΅Π½ΡΠΈΠΈ Π±ΠΎΠ»Π΅Π·Π½ΠΈ.