Kinin B1 and B2 Receptors Contribute to Cisplatin-Induced Painful Peripheral Neuropathy in Male Mice

Abstract

Cisplatin is the preferential chemotherapeutic drug for highly prevalent solid tumours. However, its clinical efficacy is frequently limited due to neurotoxic effects such as peripheral neuropathy. Chemotherapy-induced peripheral neuropathy is a dose-dependent adverse condition that negatively impacts quality of life, and it may determine dosage limitations or even cancer treatment cessation. Thus, it is urgently necessary to identify pathophysiological mechanisms underlying these painful symptoms. As kinins and their B₁ and B₂ receptors contribute to the development of chronic painful conditions, including those induced by chemotherapy, the contribution of these receptors to cisplatin-induced peripheral neuropathy was evaluated via pharmacological antagonism and genetic manipulation in male Swiss mice. Cisplatin causes painful symptoms and impaired working and spatial memory. Kinin B₁ (DALBK) and B₂ (Icatibant) receptor antagonists attenuated some painful parameters. Local administration of kinin B₁ and B₂ receptor agonists (in sub-nociceptive doses) intensified the cisplatin-induced mechanical nociception attenuated by DALBK and Icatibant, respectively. In addition, antisense oligonucleotides to kinin B₁ and B₂ receptors reduced cisplatin-induced mechanical allodynia. Thus, kinin B₁ and B₂ receptors appear to be potential targets for the treatment of cisplatin-induced painful symptoms and may improve patients' adherence to treatment and their quality of life.

Keywords: CIPN; allodynia; bradykinin; chemotherapy; neuropathic pain.

Figure 1

Characterization of cisplatin-induced painful peripheral neuropathy. (A) Male Swiss mice were treated intraperitoneally (i.p.) with cisplatin (2.3 mg/kg) or its vehicle (10 mL/kg) from day 0 onwards every other day (days 0, 2, 4, 6, 8, and 10) for a total of 10 days to induce the peripheral neuropathy experimental model. The mice’s mechanical PWT was assessed between the treatment days until 30 days after the first cisplatin administration (B). Cold sensitivity was evaluated on the 5th, 11th, 18th, and 25th days after the first cisplatin administration (C). B1 denotes baseline values measured before the first cisplatin or vehicle dose. Data are expressed as the mean + SEM (n = 6/group) and were analysed by two-way ANOVA followed by Bonferroni post hoc test. ^## p < 0.01 and ^### p < 0.001 when compared to the vehicle group. The arrows represent the days of cisplatin or vehicle administration. PWT: paw withdrawal threshold.

Figure 2

Treatment with cisplatin induced spontaneous nociceptive behaviours and impaired working and spatial memory in mice. Male Swiss mice were treated intraperitoneally (i.p.) with cisplatin (2.3 mg/kg) or its vehicle (10 mL/kg) from day 0 onwards every other day for a total of 10 days to induce the peripheral neuropathy experimental model. On the 11th day, the mice were subjected to behavioural tests. Spontaneous nociception was evaluated by voluntary wheel activity (A) and nesting behaviour (B). The cognitive function was evaluated by NORPT. The discrimination index demonstrates the preference for the novel object (C) and the total interaction time with the novel or familiar object (D). The symbols on the bars indicate individual values for each animal. ^# p < 0.05 and ^## p < 0.01 vs. vehicle group. Data are expressed as the mean + SEM (n = 6/group) and were analysed by an unpaired two-tailed Student’s t-test, except (B) nesting score (n = 6/group) data, which are expressed as 25th and 75th percentiles (interquartile range) and were analysed by one-tailed Mann–Whitney test.

Figure 3

Therapeutic effect of kinin B₁ and B₂ receptor antagonists on cisplatin-induced mechanical allodynia. (A) Therapeutic protocol: Male Swiss mice were treated intraperitoneally (i.p.) with cisplatin (2.3 mg/kg) every 48 h for 10 days. On the 11th day after the first cisplatin dose, the animals received a single administration of DALBK or SSR240612 (150 nmol/kg, i.p., peptide and non-peptide kinin B₁ receptor antagonist, respectively), Icatibant or FR173657 (100 nmol/kg, i.p., peptide and non-peptide kinin B₂ receptor antagonist, respectively), or vehicle (10 mL/kg, i.p.). Time–response curve for mechanical allodynia after treatment with DALBK or Icatibant (B) and SSR240612 or FR173657 (C). Baseline 1 (B1) values were measured before the first cisplatin dose. Baseline 2 (B2) values were measured on the 11th day after the first cisplatin dose and before the treatments. The symbols on the bars indicate individual values for each animal. ^# p < 0.05, ^## p < 0.01 and ^### p < 0.001 vs. B1 values. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. cisplatin plus vehicle group. Data are expressed as the mean + SEM (n = 6/group) and were analysed by two-way ANOVA followed by the Bonferroni post hoc test. PWT: paw withdrawal threshold.

Figure 4

Preventive effect of kinin B₁ and B₂ receptor antagonists on cisplatin-induced mechanical allodynia. (A) Preventive protocol: Male Swiss mice were treated concomitantly by intraperitoneal (i.p.) injections with cisplatin (2.3 mg/kg) plus peptide kinin B₁ (DALBK, 150 nmol/kg, i.p.) or B₂ (Icatibant, 100 nmol/kg, i.p.) receptor antagonists or vehicle (10 mL/kg, i.p.) every 48 h for 10 days. Time–response curve for PWT throughout the treatment with cisplatin plus DALBK or Icatibant (B). Baseline (B) values were measured before the first cisplatin dose. The symbols on the bars indicate individual values for each animal. ^# p < 0.05 and ^## p < 0.01 vs. B1 values. * p < 0.05 and ** p < 0.01 vs. cisplatin plus vehicle group. Data are expressed as the mean + SEM (n = 6/group) and were analysed by two-way ANOVA followed by the Bonferroni post hoc test. PWT: paw withdrawal threshold.

Figure 5

Sub-nociceptive doses of B₁ and B₂ receptor agonists intensified cisplatin-induced mechanical nociception. (A) Male Swiss mice were treated intraperitoneally (i.p.) with cisplatin (2.3 mg/kg) every 48 h for 10 days. On the 11th day after the first cisplatin injection, the animals were treated with sub-nociceptive doses of agonists kinin B₁ (DABk; 3 nmol/paw, i.pl.) or B₂ (Bk; 1 nmol/paw, i.pl.) receptor or vehicle (20 μL PBS/paw, intraplantar, i.pl.) via the intraplantar route. PWT (B,C) was assessed from 0.5 h up to 3 h after injection of sub-nociceptive doses of the agonists. Baseline 1 (B1) values were measured before cisplatin or vehicle administration. Baseline 2 (B2) values were measured on the 11th day after the first cisplatin dose and before the treatments. The symbols on the bars indicate individual values for each animal. ^### p < 0.001 vs. Veh plus Veh group. ^&& p < 0.01 vs. cisplatin plus Veh. Data are expressed as the mean + SEM (n = 6/group) and were analysed by two-way ANOVA followed by the Bonferroni post hoc test, except for the DABk effect on mechanical allodynia enhanced (Student’s t-test). Veh: vehicle; PBS: phosphate-buffered saline; PWT: paw withdrawal threshold.

Figure 6

Kinin B₁ and B₂ receptor antagonists prevent the enhancement of B₁ and B₂ receptor agonist-induced mechanical nociception in cisplatin-treated mice. (A) Male Swiss mice were treated intraperitoneally (i.p.) with cisplatin (2.3 mg/kg) every 48 h for 10 days. On the 11th day after the first cisplatin injection, the animals received a single administration of DALBK (150 nmol/kg, i.p.), Icatibant (100 nmol/kg, i.p.), or vehicle (10 mL/kg, intraperitoneal, i.p.). After 0.5 h, sub-nociceptive doses of the respective agonists, i.e., DABk (3 nmol/paw, i.pl.) or Bk (1 nmol/paw, i.pl.), or vehicle (20 μL PBS/paw, intraplantar, i.pl.) were administered via the intraplantar route. PWT (B,C) was assessed from 0.5 h up to 3 h after the treatments. Baseline 1 (B1) values were measured before cisplatin or vehicle administration. Baseline 2 (B2) values were measured on the 11th day after the first cisplatin dose and before the treatments. The symbols on the bars indicate individual values for each animal. ^### p < 0.001 vs. Veh plus Veh group. ^&& p < 0.01 vs. cisplatin plus Veh. *** p < 0.001 vs. cisplatin plus Veh. ^$$$ p < 0.001 vs. cisplatin plus DABk/Bk group. Data are expressed as the mean + SEM (n = 6/group) and were analysed by two-way ANOVA followed by the Bonferroni post hoc test, except for the DABk effect on mechanical allodynia (Student’s t-test). Veh: vehicle; PBS: phosphate-buffered saline; PWT: paw withdrawal threshold.

Figure 7

Antisense oligonucleotides for kinin B₁ and B₂ receptors relieved cisplatin-caused mechanical allodynia. (A) Male Swiss mice were treated by intraperitoneal (i.p.) injections with cisplatin (2.3 mg/kg) every 48 h for 10 days. Mice were treated with antisense oligonucleotides targeting kinin B₁ and B₂ receptors, and nonsense control was administered via the intrathecal (i.t.) route for three consecutive days every 12 h and 1 h before assessment of mechanical allodynia on the 14th day after induction of peripheral neuropathy by cisplatin. (B) Mechanical allodynia was evaluated on the 14th day after cisplatin-induced peripheral neuropathy. B1 values were measured before the first cisplatin dose. B2 values were measured on the 11th day after the first cisplatin dose and before the treatments. Results are presented as mean + SEM (n = 5–6/group). The symbols on the bars indicate individual values for each animal. ^### p < 0.001 compared to baseline threshold (B1). ** p < 0.01; *** p < 0.001 compared to the nonsense group. Two-way ANOVA repeated measures followed by Bonferroni’s post hoc test. PWT: paw withdrawal threshold.