The Role of P-Glycoprotein in Decreasing Cell Membranes Permeability during Oxidative Stress

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Alexey V. Shchulkin^1,a*, Yulia V. Abalenikhina¹, Pelageya D. Erokhina¹, Ivan V. Chernykh¹, and Elena N. Yakusheva¹

¹Ryazan State Medical University, 390026 Ryazan, Russia

^* To whom correspondence should be addressed.

Received July 15, 2020; Revised September 3, 2020; Accepted September 3, 2020

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P-Glycoprotein (P-gp) is one of the most clinically significant representatives of the ABC transporter superfamily due to its participation in the transport of biotic components and xenobiotics across the plasma membrane. It is known that various chemicals, environmental factors, and pathological processes can affect P-gp activity and expression. In this study, we investigated the role of P-gp in limiting the cell membrane permeability during oxidative stress. Human adenocarcinoma colon cells (Caco-2) overexpressing P-gp were cultured for 72 h in the medium containing hydrogen peroxide (0.1-50 µM). The transport of the P-gp substrate fexofenadine was evaluated in a special Transwell system. The amounts of P-gp and Nrf2 transcription factor were analyzed by the enzyme-linked immunosorbent assay. The concentration of SH-groups in proteins and the contents of lipid peroxidation products and protein carbonyl derivatives were determined spectrophotometrically. Hydrogen peroxide at a concentration of 0.1-5 µM did not significantly affect the studied parameters, while incubation with 10 µM H₂O₂ decreased in the level of SH groups in cell lysates and increased in the amount of Nrf2 in the cell lysates. Nrf2, in its turn, mediated an increase in the content and activity of the P-gp transporter, thus limiting the increasing permeability of the cell membrane. Hydrogen peroxide at a concentration of 50 µM promoted oxidative stress, which was manifested as a decrease in the content of SH-groups, increase in the concentration of lipid peroxidation products and protein carbonyl derivatives, and decrease in the P-gp level, which led to a significantly increased permeability of the plasma membrane. These results show that the transport and protective roles of P-gp, in particular, reduction of the cell membrane permeability, are affected by the intensity of oxidative stress and can be manifested only if the extent of membrane damage is insignificant.

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KEY WORDS: P-glycoprotein (P-gp), Nrf2, oxidative stress, permeability of biological membranes, hydrogen peroxide cytotoxicity, oxidative damage of lipids and proteins, reduced thiols

DOI: 10.1134/S0006297921020085

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