Activation of TRPV1 by capsaicin induces functional Kinin B₁ receptor in rat spinal cord microglia

Background: The kinin B₁ receptor (B₁R) is upregulated by pro-inflammatory cytokines and oxydative stress, which are enhanced by transient receptor potential vanilloid subtype 1 (TRPV1) activation. To examine the link between TRPV1 and B₁R in inflammatory pain, this study aimed to determine the ability of TRPV1 to regulate microglial B₁R expression in the spinal cord dorsal horn, and the underlying mechanism.Methods: B₁R expression (mRNA, protein and binding sites) was measured in cervical, thoracic and lumbar spinal cord in response to TRPV1 activation by systemic capsaicin (1-50 mg/kg, s.c) in rats pre-treated with TRPV1 antagonists (capsazepine or SB-366791), the antioxidant N-acetyl-L-cysteine (NAC), or vehicle. B₁R function was assessed using a tail-flick test after intrathecal (i.t.) injection of a selective B₁R agonist (des-Arg⁹-BK), and its microglial localization was investigated by confocal microscopy with the selective fluorescent B₁R agonist, [N^α-bodipy]-des-Arg⁹-BK. The effect of i.t. capsaicin (1 μg/site) was also investigated.Results: Capsaicin (10 to 50 mg/kg, s.c.) enhanced time-dependently (0-24h) B₁R mRNA levels in the lumbar spinal cord; this effect was prevented by capsazepine (10 mg/kg, i.p.; 10 μg/site, i.t.) and SB-366791 (1 mg/kg, i.p.; 30 μg/site, i.t.). Increases of B₁R mRNA were correlated with IL-1β mRNA levels, and they were significantly less in cervical and thoracic spinal cord. Intrathecal capsaicin (1 μg/site) also enhanced B₁R mRNA in lumbar spinal cord. NAC (1 g/kg/d × 7 days) prevented B₁R up-regulation, superoxide anion production and NF-kB activation induced by capsaicin (15 mg/kg). Des-Arg⁹-BK (9.6 nmol/site, i.t.) decreased by 25-30% the nociceptive threshold at 1 min post-injection in capsaicin-treated rats (10-50 mg/kg) while it was without effect in control rats. Des-Arg⁹-BK-induced thermal hyperalgesia was blocked by capsazepine, SB-366791 and by antagonists/inhibitors of B₁R (SSR240612, 10 mg/kg, p.o.), glutamate NMDA receptor (DL-AP5, 10 μg/site, i.t.), substance P NK-1 receptor (RP-67580, 10 μg/site, i.t.) and nitric oxide synthase (L-NNA, 10 μg/site, i.t.). The B₁R fluorescent agonist was co-localized with an immunomarker of microglia (Iba-1) in spinal cord dorsal horn of capsaicin-treated rats.Conclusion: This study highlights a new mechanism for B₁R induction via TRPV1 activation and establishes a link between these two pro-nociceptive receptors in inflammatory pain.