Regulation of hepatic transporters OATP1A2 and OATP1B1 by the action of nitric oxide (II)

Nitric oxide II (NO) is a signaling molecule that has a wide range of physiological effects, including the regulation of gastrointestinal processes. The liver actively expresses the clinically significant transporters OATP1A2 and OATP1B1, which are involved in the influx of biologically active and medicinal substances. That is why it seems relevant to determine the pathways of regulation of hepatic transporters under the influence of NO. Aim. To study the effect of NO on the relative amount and expression of the transporters OATP1A2 and OATP1B1 in vitro in HepG2 cells. Materials and methods. The study was performed on a culture of HepG2 cells, which were cultured in 6-well plates at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) with a high glucose content (4500 mg/l) containing L-glutamine (4 mM), 10% fetal bovine serum, 100 U/ml penicillin and 100 mg/ml streptomycin (all components from Sigma-Aldrich, Germany). S-nitrosoglutathione (Sigma-Aldrich, Germany) was added to the culture medium at concentrations of 1, 10, 50, 100 and 500 µM, incubated for 24 and 72 hours. Water for injection (solvent) was added to control cells in an equivalent volume S-nitrosoglutathione). The relative amounts of OATP1A2 and OATP1B1 proteins were assessed by Western blot, and the expression of SLCO1A2 and SLCO1B1 by real-time PCR. The results of the study. In the course of this study, it was shown that the addition of S-nitrosoglutathione in the concentration range of 10-500 μM and exposure duration of 24 and 72 hours causes an increase in the intracellular level of nitric oxide metabolites, which indicates the adequacy of the use of this NO donor. At the same time, under the influence of NO, there was an increase in the relative amount of the studied transporters - OATP1A2 at an exposure period of 24 hours and S-nitrosoglutathione concentrations of 50 and 100 μM, OATP1B1-24 and 72 hours, at concentrations of 10-500 μM, a similar trend was noted for the expression genes SLCO1A2 and SLCO1B1. Conclusion. The NO donor - S-nitrosoglutathione causes an increase in the relative amount of OATP family transporters - OATP1A2 and OATP1B1, due to increased expression of the SLCO1A2 and SLCO1B1 genes, in vitro in HepG2 cells.

HepG2, S-nitrosoglutathione, transporter protein OATP1А2/OATP1B1, SLCO1А2, SLCO1B1

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