Ginsenosides act as positive modulators of P2X4 receptors

Kshitija Dhuna, Matthew Felgate, Stefan Bidula, Samuel Walpole, Lucka Bibic, Brett Cromer, Jesus Angulo, Julie Sanderson, Martin Stebbing, Leanne Stokes (Lead Author)

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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.

Original languageEnglish
Pages (from-to)210-221
Number of pages12
JournalMolecular Pharmacology
Issue number2
Early online date13 Dec 2018
Publication statusPublished - Jan 2019


  • ATP receptors
  • P2X receptors
  • Nucleotides
  • Docking proteins
  • Gene editing/CRISPR
  • Drug discovery

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