Modulation of retinal blood flow by kinin B 1 receptor in Streptozotocin-diabetic rats

a b s t r a c t The vasoactive kinin B 1 receptor (B 1 R) is overexpressed in the retina of diabetic rats in response to hyperglycemia and oxidative stress. The aim of the present study was to determine whether B 1 R could contribute to the early retinal blood flow changes occurring in diabetes. Male Wistar rats were rendered diabetic with a single i.p. injection of Streptozotocin (STZ) and studied 4 days or 6 weeks after diabetes induction. The presence of B 1 R in the retina was confirmed by Western blot. The impact of oral administration of the B 1 R selective antagonist SSR240612 (10 mg/kg) was measured on alteration of retinal perfusion in awake diabetic rats by quantitative autoradiography. Data showed that B 1 R was upregulated in the STZ-diabetic retina at 4 days and 6 weeks. Retinal blood flow was not altered in 4-day diabetic rats compared with age-matched controls but was significantly decreased following SSR240612 treatment. In 6-week diabetic rats, retinal blood flow was markedly reduced compared to control rats and SSR240612 did not further decrease the blood flow. These results suggest that B 1 R is upregulated in STZ-diabetic retina and has a protective compensatory role on retinal microcirculation at 4 days but not at 6 weeks following diabetes induction

Activation of TRPV1 by capsaicin induces functional Kinin B₁receptor in rat spinal cord microglia

<p>Abstract</p> <p>Background</p> <p>The kinin B₁receptor (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.</p> <p>Methods</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusion</p> <p>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.</p