Cystitis-Related Bladder Pain Involves ATP-Dependent HMGB1 Release from Macrophages and Its Downstream H2S/Cav3.2 Signaling in Mice

Shiori Hiramoto, Maho Tsubota, Kaoru Yamaguchi, Kyoko Okazaki, Aya Sakaegi, Yuki Toriyama, Junichi Tanaka, Fumiko Sekiguchi, Hiroyasu Ishikura, Hidenori Wake, Masahiro Nishibori, Huy Du Nguyen, Takuya Okada, Naoki Toyooka, Atsufumi Kawabata

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)


Cystitis-related bladder pain involves RAGE activation by HMGB1, and increased Cav3.2 T-type Ca2+ channel activity by H2S, generated by upregulated cystathionine-γ-lyase (CSE) in mice treated with cyclophosphamide (CPA). We, thus, investigated possible crosstalk between the HMGB1/RAGE and CSE/H2S/Cav3.2 pathways in the bladder pain development. Bladder pain (nociceptive behavior/referred hyperalgesia) and immuno-reactive CSE expression in the bladder were determined in CPA-treated female mice. Cell signaling was analyzed in urothelial T24 and macrophage-like RAW264.7 cells. The CPA-induced bladder pain was abolished by pharmacological inhibition of T-type Ca2+ channels or CSE, and genetic deletion of Cav3.2. The CPA-induced CSE upregulation, as well as bladder pain was prevented by HMGB1 inactivation, inhibition of HMGB1 release from macrophages, antagonists of RAGE or P2X4/P2X7 receptors, and N-acetylcysteine, an antioxidant. Acrolein, a metabolite of CPA, triggered ATP release from T24 cells. Adenosine triphosphate (ATP) stimulated cell migration via P2X7/P2X4, and caused HMGB1 release via P2X7 in RAW264.7 cells, which was dependent on p38MAPK/NF-κB signaling and reactive oxygen species (ROS) accumulation. Together, our data suggest that CPA, once metabolized to acrolein, causes urothelial ATP-mediated, redox-dependent HMGB1 release from macrophages, which in turn causes RAGE-mediated CSE upregulation and subsequent H2S-targeted Cav3.2-dependent nociceptor excitation, resulting in bladder pain.

Original languageEnglish
Issue number8
Publication statusPublished - Jul 22 2020


  • Adenosine triphosphate (ATP)
  • Cav3.2 T-type Ca2+ channel
  • cyclophosphamide (CPA)
  • cystathionine-γ-lyase (CSE)
  • high mobility group box 1 (HMGB1)
  • hydrogen sulfide (H2S)
  • interstitial cystitis/bladder pain syndrome (IC/BPS)
  • macrophage
  • reactive oxygen species (ROS)
  • receptor for advanced glycation end products (RAGE)

ASJC Scopus subject areas

  • General Medicine


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