Article
Ginsenosides act as positive modulators of P2X4 receptors
Author/s | Dhuna, K.
Felgate, M. Bidula, S.M. Walpole, Samuel Bibic, L. Cromer, B.A. Angulo Álvarez, Jesús Sanderson, J. Stebbing, M.J. Stokes, L. |
Department | Universidad de Sevilla. Departamento de Química orgánica |
Publication Date | 2019 |
Deposit Date | 2020-06-09 |
Published in |
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Abstract | We investigated the selectivity of protopanaxadiol ginsenosides from Panax ginseng acting as positive allosteric modulators on P2X receptors. ATP-induced responses were measured in stable cell lines overexpressing human ... We investigated the selectivity of protopanaxadiol ginsenosides from Panax ginseng acting as positive allosteric modulators on P2X receptors. ATP-induced responses were measured in stable cell lines overexpressing human P2X4 using a YOPRO-1 dye uptake assay, intracellular calcium measurements, and whole-cell patch-clamp recordings. Ginsenosides CK and Rd were demonstrated to enhance ATP responses at P2X4 by ∼twofold, similar to potentiation by the known positive modulator ivermectin. Investigations into the role of P2X4 in mediating a cytotoxic effect showed that only P2X7 expression in HEK-293 cells induces cell death in response to high concentrations of ATP, and that ginsenosides can enhance this process. Generation of a P2X7-deficient clone of BV-2 microglial cells using CRISPR/ Cas9 gene editing enabled an investigation of endogenous P2X4 in a microglial cell line. Compared with parental BV-2 cells, P2X7-deficient BV-2 cells showed minor potentiation of ATP responses by ginsenosides, and insensitivity to ATP 2 or ATP 1 ginsenoside-induced cell death, indicating a primary role for P2X7 receptors in both of these effects. Computational docking to a homology model of human P2X4, based on the open state of zfP2X4, yielded evidence of a putative ginsenoside binding site in P2X4 in the central vestibule region of the large ectodomain. |
Citation | Dhuna, K., Felgate, M., Bidula, S.M., Walpole, S., Bibic, L., Cromer, B.A.,...,Stokes, L. (2019). Ginsenosides act as positive modulators of P2X4 receptors. Molecular Pharmacology, 95 (2), 210-221. |
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