In vivo B1 kinin-receptor upregulation. Evidence for involvement of protein kinases and nuclear factor kappaB pathways

Abstract

1. Intradermal (i.d.) injection of cytokines, IL-1beta and TNFalpha (5 ng, 60 and 30 min prior) produces a rapid onset up-regulation of des-Arg9-BK-mediated rat paw oedema. Here we analyse the mechanisms involved in des-Arg9-BK-induced oedema in animals pre-treated with IL-1beta or TNFalpha. 2. Co-injection of anti-IL-1beta, anti-TNFalpha and anti-IL-8 (50 ng) significantly inhibited des-Arg9-BK-induced oedema in animals pre-treated with IL-1beta (65, 37 and 42%) or TNFalpha (39, 64, 25%). IL-1 receptor antagonist (IRA, 100 microg) or IL-10 (10 ng) inhibited the oedema caused by des-Arg9-BK, in rats that had received either IL-1beta (67 and 63%) or TNFalpha (46 and 35%). 3. Co-injection of the PKC inhibitors, staurosporine (10 nmol) or RO 318220 (30 nmol) inhibited des-Arg9-BK-induced paw oedema (44 and 42% for IL-1beta and, 53 and 30% for TNFalpha, respectively). Genistein (tyrosine kinase inhibitor, 2.5 mg kg-1, s.c.) or PD 098059 (MAP-kinase inhibitor, 30 nmol) produced marked inhibition of des-Arg9-BK-induced oedema (58 and 39% for IL-1beta and 31 and 35% for TNFalpha respectively). 4. The NF-kappaB inhibitors TLCK (2 mg kg-1, i.p.) and PDCT (100 mg kg-1, i.p.) significantly inhibited the oedema of des-Arg9-BK in IL-1beta (27 and 83%) or TNFalpha (28 and 80%) pre-treated animals. 5. It is concluded that up-regulation of B1 receptors modulated by IL-1beta or TNFalpha involves the release of other cytokines, activation of PKC and tyrosine kinase pathways, co-ordinated with the activation of MAP-kinase and nuclear factor kappaB, reinforcing the view that B1 receptors may exert a pivotal role in modulating chronic inflammatory processes.

Figure 1

Effect of co-injection of anti-murine antibodies anti-IL-1β (A and B), anti-TNFα (C and D) or anti-IL-8 (E and F) on des-Arg⁹-BK-induced paw oedema in rats pre-treated with IL-1β (5 ng paw⁻¹, 60 min beforehand; A, C and E) or with TNFα (5 ng paw⁻¹, 30 min beforehand; (B, D and F)). Values represent the differences between volumes (in ml) of vehicle-injected (0.1 ml of PBS solution) and drug-injected paws. Each point represents the mean±s.e.mean of 4–6 rats. In some cases the error bars are hidden within the symbols. Significantly different from control values ^*P<0.05; ^**P<0.01 (A–E, Student's unpaired t-test) (F, ANOVA followed by Dunnett's test).

Figure 2

Effect of co-injection of the recombinant human IL-1 antagonist (IRA, A and B) or of IL-10 (C and D) on des-Arg⁹-BK-induced paw oedema in rats pre-treated with IL-1β (5 ng paw⁻¹, 60 min beforehand; (A and C)) or with TNFα (5 ng paw⁻¹, 30 min beforehand; (B and D)). Values represent the differences between volumes (in ml) of vehicle-injected (0.1 ml of PBS solution) and drug-injected paws. Each point represents the mean±s.e.mean of 4–6 rats. In some cases the error bars are hidden within the symbols. Significantly different from control values ^*P<0.05; ^**P<0.01 (A–C, Student's unpaired t-test) (D, ANOVA followed by Dunnett's test).

Figure 3

Effect of co-injection of staurosporine (A and B) or of RO 319220 (C and D) on des-Arg⁹-BK-induced paw oedema in rats pre-treated with IL-1β (5 ng paw⁻¹, 60 min beforehand (A and C)) or with TNFα (5 ng paw⁻¹, 30 min beforehand (B and D)). Values represent the differences between volumes (in ml) of vehicle-injected (0.1 ml of PBS solution) and drug-injected paws. Each point represents the mean±s.e.mean of 4–6 rats. In some cases the error bars are hidden within the symbols. Significantly different from control values ^*P<0.05; ^**P<0.01 (Student's unpaired t-test).

Figure 4

Effect of systemic treatment with genistein (A and B) or with PD 098059 (C and D) on des-Arg⁹-BK-induced paw oedema in rats pre-treated with IL-1β (5 ng paw⁻¹, 60 min beforehand (A and C)) or with TNFα (5 ng paw⁻¹, 30 min beforehand (B and D)). Values represent the differences between volumes (in ml) of vehicle-injected (0.1 ml of PBS solution) and drug-injected paws. Each point represents the mean±s.e.mean of 4–6 rats. In some cases the error bars are hidden within the symbols. Significantly different from control values ^*P<0.05; ^**P<0.01 (Student's unpaired t-test).

Figure 5

Effect of systemic treatment with TLCK (A and B) or with PDCT (C and D) on des-Arg⁹-BK-induced paw oedema in rats pre-treated with IL-1β (5 ng paw⁻¹, 60 min beforehand (A and C)) or with TNFα (5 ng paw⁻¹, 30 min beforehand (B and D)). Values represent the differences between volumes (in ml) of vehicle-injected (0.1 ml of PBS solution) and drug-injected paws. Each point represents the mean±s.e.mean of 4–6 rats. In some cases the error bars are hidden within the symbols. Significantly different from control values ^*P<0.05; ^**P<0.01 (Student's unpaired t-test).