Ginsenosides act as positive modulators of P2X4 receptors

Dhuna, Kshitija, Felgate, Matthew, Bidula, Stefan ORCID: https://orcid.org/0000-0002-3790-7138, Walpole, Samuel, Bibic, Lucka, Cromer, Brett, Angulo, Jesus ORCID: https://orcid.org/0000-0001-7250-5639, Sanderson, Julie, Stebbing, Martin and Stokes, Leanne ORCID: https://orcid.org/0000-0003-4013-6781 (2019) Ginsenosides act as positive modulators of P2X4 receptors. Molecular Pharmacology, 95 (2). pp. 210-221. ISSN 0026-895X

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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 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.

Item Type: Article
Uncontrolled Keywords: atp receptors,p2x receptors,nucleotides,docking proteins,crispr,drug discovery,pharmacology ,/dk/atira/pure/subjectarea/asjc/3000/3004
Faculty \ School: Faculty of Science > School of Pharmacy
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 20 Dec 2018 16:31
Last Modified: 21 Oct 2022 21:32
URI: https://ueaeprints.uea.ac.uk/id/eprint/69408
DOI: 10.1124/mol.118.113696

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