Kinin Receptors Sensitize TRPV4 Channel and Induce Mechanical Hyperalgesia: Relevance to Paclitaxel-Induced Peripheral Neuropathy in Mice

  • Robson Costa
  • Maíra A. Bicca
  • Marianne N. Manjavachi
  • Gabriela C. Segat
  • Fabiana Chaves Dias
  • Elizabeth S. Fernandes
  • João B. Calixto
Article
  • 401 Downloads

Abstract

Kinin B1 (B1R) and B2 receptors (B2R) and the transient receptor potential vanilloid 4 (TRPV4) channel are known to play a critical role in the peripheral neuropathy induced by paclitaxel (PTX) in rodents. However, the downstream pathways activated by kinin receptors as well as the sensitizers of the TRPV4 channel involved in this process remain unknown. Herein, we investigated whether kinins sensitize TRPV4 channels in order to maintain PTX-induced peripheral neuropathy in mice. The mechanical hyperalgesia induced by bradykinin (BK, a B2R agonist) or des-Arg9-BK (DABK, a B1R agonist) was inhibited by the selective TRPV4 antagonist HC-067047. Additionally, BK was able to sensitize TRPV4, thus contributing to mechanical hyperalgesia. This response was dependent on phospholipase C/protein kinase C (PKC) activation. The selective kinin B1R (des-Arg9-[Leu8]-bradykinin) and B2R (HOE 140) antagonists reduced the mechanical hyperalgesia induced by PTX, with efficacies and time response profiles similar to those observed for the TRPV4 antagonist (HC-067047). Additionally, both kinin receptor antagonists inhibited the overt nociception induced by hypotonic solution in PTX-injected animals. The same animals presented lower PKCε levels in skin and dorsal root ganglion samples. The selective PKCε inhibitor (εV1–2) reduced the hypotonicity-induced overt nociception in PTX-treated mice with the same magnitude observed for the kinin receptor antagonists. These findings suggest that B1R or B2R agonists sensitize TRPV4 channels to induce mechanical hyperalgesia in mice. This mechanism of interaction may contribute to PTX-induced peripheral neuropathy through the activation of PKCε. We suggest these targets represent new opportunities for the development of effective analgesics to treat chronic pain.

Keywords

Paclitaxel Peripheral neuropathy Neuropathic pain Kinins TRPV4 

Introduction

Peripheral neuropathy caused by paclitaxel (PTX) is a dose-limiting side effect characterized by painful paraesthesia of the hands and feet [1, 2]. Kinins were recently implicated in the physiopathology of PTX-induced peripheral neuropathy in mice [3]. Kinins are algogenic peptides endogenously formed by the cleavage of kininogens by kallikreins. They act on two subtypes of G protein-coupled receptors, called B1 (B1R) and B2 (B2R) receptor. Typically constitutive, B2R receptors are expressed in peripheral tissues and the central nervous system, displaying higher affinity for bradykinin (BK) and kallidin peptides and mediating most of the physiological effects of the kinins. In contrast, B1R shows higher affinity for the kinin metabolites des-Arg9-BK (DABK) and Lys-des-Arg9-BK, and it is usually absent in peripheral tissues under physiological conditions. However, B1R can be upregulated during pathological states [4, 5] and its constitutive expression was previously detected in the central nervous system [6, 7].

The contribution of both kinin receptors to the transduction of pain has been widely demonstrated, particularly in chronic painful states. Of note, B1R and B2R are suggested to be involved in both the mechanical and thermal hypersensitivities induced by PTX in mice, but little is known of the mechanisms underlying these processes [3]. Evidences from in vitro and in vivo studies have shown that the pro-algesic effects of kinins can be mediated by sensitization of transient receptor potential (TRP) channels, including the TRP vanilloid 1 (TRPV1) and TRP ankyrin 1 (TRPA1) channels [8, 9, 10, 11, 12, 13, 14]. Recently, BK was shown to sensitize the TRP vanilloid 4 (TRPV4) channel via a protein kinase C (PKC)-dependent mechanism [15], suggesting this channel might be implicated in the nociceptive actions of kinins.

TRPV4 is a cation channel sensitive to osmolarity changes and mechanical stimuli [16]. Indeed, hypotonicity induces the excitation of dorsal root ganglion (DRG) neurons by activating TRPV4 channels and increasing intracellular Ca2+ levels [17]. Inflammatory mediators have been shown to sensitize rodents to the nociceptive effects of hypotonicity, causing TRPV4-dependent overt nociception and mechanical hyperalgesia [17, 18, 19]. TRPV4 is also involved in PTX-induced peripheral neuropathy, by mediating both mechanical hyperalgesia and increased sensitivity to hypotonicity [20, 21, 22, 23].

Despite the well-characterized involvement of kinin receptors and TRPV4 channels in PTX-induced peripheral neuropathy in rodents, the downstream pathways to kinin receptor activation, as well as the sensitizers of the TRPV4 channel involved in the maintenance of PTX-induced mechanical hypersensitivity, are currently unknown. Herein, we present evidence on the crosstalk between kinins and TRPV4 channels and their contribution to PTX-induced mechanical hyperalgesia in mice.

Methods

Animals

Male adult Swiss mice (8–10 weeks) were used. Animals were housed in a room with controlled temperature (22 ± 2 °C) and humidity (~60–80%) under a 12:12 h light-dark cycle (lights on at 06:00 a.m.). Food and water were provided ad libitum. Mice were randomly distributed in experimental groups (n = 6–10/group), and behavioural experiments were conducted in a blinded manner in order to reduce experimental bias. The n numbers and the intensity of noxious stimuli used in this study were the minimum necessary to achieve consistent effects. All experimental procedures were carried out in accordance with the National Institutes of Health animal care guidelines (NIH publications No. 8023) and were approved by the Ethics Committee of the Universidade Federal de Santa Catarina (protocol number PP00811). All animal studies are in compliance with the ARRIVE guidelines [24].

Mechanical Hyperalgesia Induced by Selective Kinin Receptor Agonists

In order to investigate whether TRPV4 contributes to the mechanical hyperalgesia induced by kinins, animals (n = 8/group) received either the selective kinin B2 (BK; 3 nmol/paw; 20 μl) or B1 (DABK; 30 nmol/site; 5 μl) receptor agonist by intraplantar (i.pl.) and intrathecal (i.t.) routes, respectively. Vehicle (saline)-treated animals were used as controls. Mechanical nociceptive thresholds were measured at different time points (20–240 min) following the injection of the stimuli.

The involvement of TRPV4 channels in these responses was assessed in mice pre-treated with the selective TRPV4 antagonist HC-067047, either by intraperitoneal (i.p.; 10 mg/kg, 1 h prior to the stimuli) or i.t. (3 μg/site, co-injected with the stimuli) routes. Control animals received the same volume of vehicle by i.p. (8% dimethyl sulfoxide (DMSO) plus 2% Tween 80 in saline) or i.t. (2% DMSO) routes. i.t. injections were performed as previously described [25], with minor modifications [26]. Treatment schedules are represented in Fig. 1(a, b).
Fig. 1

Schematic representation of treatments in acute protocols. a Mechanical hyperalgesia induced by the selective B2 receptor agonist bradykinin (BK; 3 nmol/paw). b Mechanical hyperalgesia induced by the selective B1 receptor agonist des-Arg9-BK (DABK; 30 pmol/site, i.t.). c Mechanical hyperalgesia induced by TRPV4 activators in animals sensitized by BK (0.3 nmol/paw)

Mechanical Hyperalgesia Induced by TRPV4 Activation in Animals Sensitized by Kinin Receptor Agonists

It was investigated whether BK or DABK could sensitize mice to the nociceptive response elicited by the TRPV4 activators, 4α-PDD and hypotonic solution. For this, 10 μl of saline (0.9% NaCl) containing either BK (0.3 nmol/paw) or DABK (20 nmol/paw) were injected intraplantarly into the right mouse hindpaw (n = 8/group). Control groups received an equal volume of saline. After 5 min, the TRPV4 agonist 4α-PDD (1 nmol/paw, 10 μl of 1% DMSO) or hypotonic solution (10 μl of deionized water) was injected into paws previously treated with the algogens. Mechanical thresholds were evaluated 15–120 min after the stimuli. Control groups received 10 μl of vehicle (1% DMSO) or isotonic solution (0.9% NaCl), respectively.

To evaluate the mechanisms implicated in the sensitizing effect of BK to hypotonicity, animals received one of the following drugs: the selective TRPV4 (HC-067047; 3 μg/paw), kinin B1 (des-Arg9-[Leu8]-bradykinin, DALBK; 3 nmol/paw), kinin B2 (HOE 140; 3 nmol/paw) or TRPV1 (SB366791; 1 nmol/paw) receptor antagonists; the non-selective cyclooxygenase (COX) 1/2 (indomethacin; 5 mg/kg, p.o.) or catecholamine release (guanethidine; 30 mg/kg, s.c.) inhibitors, to assess the involvement of prostaglandins and catecholamines, respectively; or the selective phospholipase A2 (PLA2) (PACOCF3; 1 nmol/paw), protein kinase A (PKA) (KT-5720; 3 nmol/paw), phospholipase C (PLC) (U73122; 30 pmol/paw) or PKC (GF109203X; 3 nmol/paw) inhibitors. Indomethacin and guanethidine were given 1 h prior to BK (0.3 nmol/paw). All other drugs were co-injected with BK (5 min prior to hypotonic solution injection). Treatment schedules are represented in Fig. 1(c).

Mechanical Hyperalgesia Induced by Hypotonicity in Animals Sensitized by PKCε Activation

We also evaluated whether PKCε activation sensitizes mice to hypotonicity. Thus, the selective PKCε activator ΨεRACK (0.1 μg/paw) or its vehicle (0.9% NaCl; control group) was intraplantarly injected (in 10 μl of saline) into the right mouse hindpaw (n = 8/group). After 5 min, a hypotonic solution (10 μl of deionized water) was injected into the paws previously treated with the sensitizing agent, and the mechanical thresholds were then evaluated after 15 min. Control groups received 10 μl of isotonic solution (0.9% NaCl).

Mechanical Hyperalgesia Induced by PTX in Mice

Peripheral neuropathy was induced by PTX as previously described [1] and adapted for mice [3]. Briefly, mice (n = 6–10/group) were injected with PTX (2 mg/kg, i.p.) for five consecutive days (days 0 to 4), using an injection volume of 10 ml/kg. The cumulative dose of PTX was of 10 mg/kg. Control animals received vehicle (0.9% NaCl). The development of peripheral neuropathy was assessed 7 days after the first injection of PTX by testing the mouse sensitivity to mechanical stimuli.

To assess the participation of TRPV4, kinin B1 and B2 receptors in this response, animals received either the selective TRPV4 (HC-067047; 10 mg/kg, i.p.), kinin B1 (DALBK; 150 nmol/kg, i.p.) or B2 (HOE 140; 50 nmol/kg, i.p.) receptor antagonist on day 7, and the mechanical hyperalgesia was evaluated at different time points following injection (60–360 min). Treatment schedules are represented in Fig. 2(a).
Fig. 2

Schematic representation of treatments in paclitaxel (PTX)-induced peripheral neuropathy. a Mechanical hyperalgesia induced by PTX in mice. b Overt nociception induced by hypotonicity in PTX-treated mice

Mechanical Threshold Measurements

Mechanical nociceptive thresholds were evaluated as previously described [27]. Initially, mice were individually placed in clear Plexiglas boxes (9 × 7 × 11 cm) on elevated wire-mesh platforms to allow access to the ventral surface of the right hindpaw. Animals were acclimatized to the boxes for 1 h before the initiation of the behavioural tests. Through the wire-mesh floor of the chamber, a series of eight von Frey hair monofilaments (Stöelting, Wood Dale, IL, USA), calibrated to produce incremental forces of 0.02 to 2 g, were applied to the plantar surface of the right hindpaw for a maximum of 3 s, or until the animal displayed a nocifensive response. Tests were initiated with a 0.6-g filament. In the absence of a nocifensive response, incrementally stronger filaments were used consecutively until a response was elicited. If the 0.6-g filament elicited a response, decrementally weaker filaments were used until one of them failed to cause a nocifensive response. The data collected using this up-down method were then used to calculate paw withdrawal thresholds (in g) [28]. Baseline measurements were taken before the injection of the nociceptive stimuli. Significant decreases in the paw withdrawal thresholds were interpreted as indicatives of mechanical hyperalgesia.

Overt Nociception Induced by Hypotonicity in PTX-Treated Mice

Overt nociception was induced as described previously [20]. Briefly, 7 days after the first PTX treatment, animals (n = 10) were individually placed into inverted glass cylinders of 20 cm in diameter, for at least 30 min, in order to acclimatize them to the experimental environment. Then, 20 μl of hypotonic (deionized water) or isotonic (0.9% NaCl; control) solutions was injected intraplantarly, in both PTX- and vehicle-treated mice, into the right mouse hindpaw, using a 500-μl insulin syringe (30-gauge needle). Mice were then observed for 5 min. The amount of time spent shaking, licking, flinching or elevating the injected paw was measured with a chronometer and considered as an indicative of overt nociception.

In order to assess the contribution of TRPV4, kinin B1 and B2 receptors to the increased sensitivity to hypotonicity in PTX-treated mice, animals received either HC-067047 (3 μg/paw), DALBK (150 nmol/kg, i.p.) or HOE 140 (50 nmol/kg, i.p.), respectively. We also verified the involvement of PKCε in this response by using the selective peptide PKCε inhibitor εV1–2 (9 μg/paw). HC-067047 was co-injected with hypotonic solution 7 days after the first injection of PTX. DALBK, HOE 140 or the εV1–2 peptide was given twice a day (every 12 h) for 6 days, starting at the time of the first PTX treatment. Eight to twelve animals were used per group in each experiment. Treatment schedules are represented in Fig. 2(b).

Membrane and Cytosolic Extract Preparation

To evaluate the contribution of kinin B1R and B2R to the increased levels of PKCε in PTX-treated mice, neuropathic mice received either the selective B1 DALBK (150 nmol/kg, i.p., n = 6–8) or B2 HOE 140 (50 nmol/kg, i.p., n = 6) receptor antagonists, given twice a day (every 12 h) for 6 days, starting at the time of the first PTX treatment. Vehicle-treated mice were used as controls (n = 6). Seven days after the first PTX treatment, animals (under isoflurane anaesthesia) were killed by decapitation, and the hindpaw skins and L1 to L6 DRGs were collected. PKCε levels were assessed as previously described [29], with minor modifications. Hindpaw skin samples were homogenized in 500 μl of ice-cold buffer A (10 mM HEPES; pH 7.4, 2 mM MgCl2, 10 mM KCl, 1 mM EDTA, 1 mM phenylmethylsulfonyl fluoride, 50 mm sodium fluoride, 10 mm β-glycerophosphate, 1 mM dithiothreitol, 10 μg/ml leupeptin, 10 μg/ml pepstatin A, 10 μg/ml aprotinin, 1 mM sodium orthovanadate). Homogenates were vigorously shaken and chilled on ice for 15 min and then centrifuged at 14,000 rpm for 45 min, at 4 °C. Supernatants (cytosolic fraction) were stored at −70 °C until further use. For the obtention of membrane extracts, the resulting pellets were washed twice in ice-cold PBS and suspended in 200 μl of buffer A containing Triton X-100 (1%). Samples were sonicated for 5 min, centrifuged at 14,000 rpm, at 4 °C, for 45 min, and the supernatant was collected and stored at −70 °C until further use. Dorsal root ganglion tissue samples from L1 to L6 segments were homogenized in 50 μl of RIPA buffer containing protease inhibitors. The lysates were centrifuged twice at 14,000g for 10 min, at 4 °C, and the supernatant was collected and stored at −70 °C until further use.

Protein concentrations were determined by spectrophotometry (NanoDrop Spectrophotometer ND-1000; Thermo Scientific, Rockford, IL, EUA). Samples were then diluted in 5× Laemmli buffer (25% of 62.5 mM Tris-HCl (pH 6.8), 2% glycerol, 0.01% SDS and bromophenol blue) containing 5% β-mercaptoethanol, adjusted to the same amount of protein and boiled at 100 °C for 5 min.

Western Blotting Assay

Fifteen micrograms of protein were loaded per lane and electrophoretically separated using 12% denaturing SDS polyacrylamide gel electrophoresis (PAGE). Proteins were then transferred to nitrocellulose membranes using a Mini Trans-Blot Cell System (Bio-Rad Laboratories, Inc., Hercules, CA, USA) following the manufacturer’s protocol. Membranes were blocked using 10% BSA in 0.05% TBST solution for 1 h, at 4 °C, and probed with the following antibodies: mouse anti-β-actin (sc-81178, 1:1000) and rabbit anti-PKCε (sc-214, 1:1000), both from Santa Cruz Technology Biotechnology, Inc. (Dallas, TX, USA), diluted in blocking buffer and incubated at 4 °C, overnight. Following incubation, membranes were washed and incubated with specific secondary antibodies conjugated to horseradish peroxidase (1:25.000; Cell Signaling Technology, Danvers, MA, USA). Immunocomplexes were visualized using SuperSignal West Femto Chemiluminescent Substrate Detection System (Thermo Fischer Scientific, Rockford, IL, USA). Protein levels were quantified by optical density using ImageJ Software®, and densitometric values were normalized as the ratio to β-actin in arbitrary units (AU). Results are expressed as a percentage of PKCε expression in relation to the control group.

Drugs

The following drugs were used: paclitaxel (PTX; Dosa S.A., Buenos Aires, Argentina); 4α-phorbol 12,13-didecanoate (4α-PDD), bradykinin (BK), des-Arg9-bradykinin (DABK), des-Arg9-[Leu8]-bradykinin (DALBK), guanethidine, HC-067047, indomethacin and SB366791 (Sigma Chemical, Saint Louis, MO, USA); GF109203X, KT-5720, PACOCF3 and U73122 (Tocris Bioscience, Bristol, UK); and εV1–2 (Glu-Ala-Val-Ser-Leu-Lys-Pro-Thr) and ΨεRACK (His-Asp-Ala-Pro-IIe-Gly-Tyr-Asp) (GenScript USA, Inc., Piscataway, NJ, USA). PTX stock solution (6 mg/ml in Cremophor EL®) was diluted in saline (0.9% NaCl) to a concentration of 0.2 mg/ml (injection solution). 4α-PDD and HC-067047 were diluted in 1% dimethyl sulfoxide (DMSO) in saline. Indomethacin was diluted in 5% Na2CO3. SB366791, GF109203X, KT-5720, PACOCF3 and U73122 stock solutions (10−3 M) were prepared in ethanol. For in vivo experiments, the final concentration of ethanol did not exceed 1%. All other drugs were dissolved in saline. The doses of TRPV4 and kinin B1 and B2 receptor antagonists were selected based on previous studies [3, 30].

Statistical Analysis

Results are presented as the mean ± SEM of 6–10 animals for each experimental group. Statistical comparisons were performed by two-way ANOVA followed by the Bonferroni post test, one-way ANOVA followed by the Newman–Keuls post test or Student’s t test, in GraphPad Prism software, version 5.01. P values <0.05 were considered significant. Power analysis was performed in order to calculate the size of the experimental groups.

Results

TRPV4 Mediates the Mechanical Hyperalgesia Induced by Kinin Receptor Agonists

As shown in Fig. 3a, b, both the i.pl. injection of BK (3 nmol/paw) and the i.t. injection of DABK (30 nmol/site) induced significant decreases in the mechanical withdrawal thresholds when compared with the saline-injected group. The systemic treatment with HC-067047 (10 mg/kg, i.p., 1 h prior) markedly inhibited BK-induced responses (Fig. 3a). Similarly, DABK-induced mechanical hyperalgesia was significantly attenuated by the co-treatment with this compound (3 μg/site, Fig. 3b).
Fig. 3

TRPV4 mediates the mechanical hyperalgesia induced by kinins in mice. a Effect of the selective TRPV4 channel antagonist HC-067047 (10 mg/kg, i.p., 1 h) on bradykinin (BK; 3 nmol/paw)-induced mechanical hyperalgesia in mice. b Effect of the selective TRPV4 channel antagonist HC-067047 (3 μg/site, co-injected) on mechanical hyperalgesia induced by the intrathecal (i.t.) injection of des-Arg9-BK (DABK; 30 nmol/site) in mice. Each group represents the mean of eight animals, and the error bars indicate the SEM. *P < 0.05, significantly different from saline-injected mice. #P < 0.05, significantly different from mice injected with BK (a) or DABK (b). Two-way ANOVA followed by the Bonferroni post test. BL baseline withdrawal threshold

BK Sensitizes Mice to TRPV4 Activators Causing Mechanical Hyperalgesia

Neither 4α-PDD (1 nmol/paw) nor hypotonic solution (10 μl of deionized water) i.pl. injection in saline pre-injected mice caused significant alterations in the mechanical threshold within 15 min after their injections, when compared with control groups (Fig. 4a, b). However, when BK (0.3 nmol/paw) was injected 5 min prior to either 4α-PDD (Fig. 4a) or hypotonic solution (Fig. 4b), a significant reduction was observed in the mechanical thresholds to von Frey filament stimuli. The sensitizing effect of BK to 4α-PDD (data not shown) or hypotonic solution (Fig. 4c) was observed within the first 15 min and lasted for 60 min following the injection of TRPV4 activators. The treatment with either the selective B2R (HOE 140; 3 nmol/paw) or TRPV4 (HC-067047; 3 μg/paw), but not the B1R antagonist (DALBK; 3 nmol/paw), significantly prevented the sensitizing effect of BK to hypotonicity (Table 1). Additionally, pre-injection of the selective B1R agonist DABK (20 nmol/paw) was not able to sensitize mice to hypotonicity (data not shown).
Fig. 4

Bradykinin sensitizes mice to TRPV4 activators and causes mechanical hyperalgesia. a, b Effect of a sub-effective dose of bradykinin (BK; 0.3 nmol/paw) on the plantar mechanical threshold after the injection of TRPV4 activators, a 4α-PDD (1 nmol/paw) or b hypotonic solution (Hypo; 10 μl of deionized water per paw). BK was injected 5 min before TRPV4 activators (or their vehicles). Plantar mechanical threshold was evaluated 15 min after the injection of TRPV4 activators (or their vehicles). Iso isotonic solution. c Time response profile of the experiments represented in b. Each group represents the mean of eight animals, and the error bars indicate the SEM. *P < 0.05, significantly different from saline-injected groups. a, b One-way ANOVA followed by the Newman–Keuls post test. c Two-way ANOVA followed by the Bonferroni post test. BL baseline withdrawal threshold

Table 1

Effect of different classes of drugs on the sensitizing effect of bradykinin to hypotonicity

Drugs and doses

Mechanism

Mechanical threshold (g)

Inhibition (%)

Basal

Hypotonicity (10 μl of deionized water)

Saline

BK (0.3 nmol/paw)

+ Vehicle

+ Drug

HC-067047 (3 μg/paw)

TRPV4 antagonist

0.88 ± 0.12

0.66 ± 0.10

0.12 ± 0.05*

0.54 ± 0.16#

78 ± 30%

HOE 140 (3 nmol/paw)

B2R antagonist

0.87 ± 0.11

0.66 ± 0.11

0.15 ± 0.04*

0.56 ± 0.09#

80 ± 18%

DALBK (3 nmol/paw)

B1R antagonist

0.87 ± 0.11

0.66 ± 0.11

0.15 ± 0.04*

0.14 ± 0.07*

N.S.

Indomethacin (5 mg/kg, p.o.)

COX1/2 inhibitor

0.87 ± 0.11

0.62 ± 0.05

0.13 ± 0.05*

0.18 ± 0.05*

N.S.

Guanethidine (30 mg/kg, s.c.)

Sympathetic amine release inhibitor

0.89 ± 0.15

0.76 ± 0.13

0.15 ± 0.04*

0.21 ± 0.09*

N.S.

PACOCF3 (1 nmol/paw)

PLA2 inhibitor

1.02 ± 0.10

0.75 ± 0.05

0.24 ± 0.08*

0.21 ± 0.07*

N.S.

KT-5720 (3 nmol/paw)

PKA inhibitor

0.98 ± 0.05

0.76 ± 0.13

0.22 ± 0.09*

0.27 ± 0.08*

N.S.

SB366791 (1 nmol/paw)

TRPV1 antagonist

0.84 ± 0.13

0.62 ± 0.08

0.19 ± 0.10*

0.26 ± 0.11*

N.S.

U73122 (30 pmol/paw)

PLC inhibitor

1.02 ± 0.13

0.77 ± 0.06

0.24 ± 0.08*

0.65 ± 0.12#

77 ± 22%

GF109203X (3 nmol/paw)

PKC inhibitor

0.95 ± 0.10

0.70 ± 0.06

0.20 ± 0.05*

0.48 ± 0.10#

56 ± 20%

N.S. no significant inhibition.*P < 0.05, significantly different from saline-injected mice. #P < 0.05, significantly different from BK + vehicle group. One-way ANOVA followed by the Newman–Keuls post-test.

The Sensitizing Effect of BK to Hypotonicity Is Dependent on PLC and PKC Activation

As depicted in Table 1, treatment with the selective PLC (U73122; 30 pmol/paw) or PKC (GF00109203X; 3 nmol/paw) inhibitors significantly attenuated the mechanical hyperalgesia induced by hypotonicity in animals pre-injected with BK (0.3 nmol/paw). However, neither the treatment with the selective PLA2 (PACOCF3; 1 nmol/paw) nor the PKA (KT-5720; 3 nmol/paw) inhibitor prevented this response (Table 1). Similarly, neither the i.pl injection with the selective TRPV1 antagonist SB366791 (1 nmol/paw) nor the systemic treatment with the non-selective COX 1/2 (indomethacin; 5 mg/kg, p.o., 1 h) or the catecholamine release (guanethidine; 30 mg/kg, s.c., 1 h) inhibitor was able to interfere with the BK-induced sensitization to hypotonicity (Table 1).

PTX-Induced Mechanical Hyperalgesia Is Similarly Reduced by Both Kinin Receptors and TRPV4 Antagonism

The administration of the selective TRPV4 channel antagonist HC-067047 (10 m/kg, i.p.) inhibited PTX-induced mechanical hyperalgesia for up to 4 h after its administration, when evaluated 7 days after the first PTX treatment (Fig. 5a). A similar profile was observed in animals treated with either the kinin B1R (DALBK; 150 nmol/kg, i.p.) or B2R (HOE 140; 50 nmol/kg, i.p.) antagonists (Fig. 5a).
Fig. 5

Kinin receptor activation sensitizes TRPV4 in neuropathic mice. a Effect of the selective kinin B1 (DALBK; 150 nmol/kg, i.p.) or B2 (HOE 140; 50 nmol/kg, i.p.) receptor or TRPV4 channel (HC-047067, 10 mg/kg, i.p.) antagonists on PTX-induced mechanical hyperalgesia in mice. b Overt nociception induced by hypotonic solution (20 μl of deionized water per paw) in PTX-treated animals. c Effect of selective kinin B1 (DALBK) or B2 (HOE 140) receptor antagonists on the overt nociception induced by hypotonic solution in PTX-treated animals. Each group represents the mean of 6 (a) or 10 (b, c) animals, and the error bars indicate the SEM. *P < 0.05, significantly different from control (a, c) or control + isotonicity (b) groups. #P < 0.05, significantly different from the control + hypotonicity group. a Two-way ANOVA followed by the Bonferroni post test. b, c One-way ANOVA followed by the Newman–Keuls post test. BL baseline withdrawal threshold

Kinin Receptors Mediate Hypotonicity-Induced Overt Nociception in PTX-Treated Mice

PTX-treated mice exhibited increased overt nociception following the i.pl. injection of hypotonic solution (10 μl of deionized water) when compared with vehicle-treated animals (Fig. 5b). This response was prevented by the co-treatment with the selective TRPV4 antagonist HC-067047 (3 μg/paw) (data not shown). Similarly, mice systemically treated with either the selective kinin B1R (DALBK; 100 nmol/kg) or B2R (HOE 140; 50 nmol/kg) antagonists (Fig. 5c) did not exhibit significant overt nociception.

Kinin Receptor Antagonism Inhibited the Increased Levels of PKCε in PTX-Treated Mice

PKCε levels were found to be increased in plantar skin membrane and cytosolic preparations (Fig. 6a) and DRG total extracts (Fig. 6b) obtained from PTX-treated animals. Importantly, the treatment with the kinin B1R (DALBK; 100 nmol/kg, i.p.) and B2R (HOE 140; 50 nmol/kg, i.p.) antagonists significantly prevented the increased levels of PKCε in all the evaluated tissues obtained from neuropathic animals (Fig. 6a, b).
Fig. 6

Kinin receptor antagonists inhibited the increased levels of PKCε in paclitaxel-treated mice. a Levels of PKCε in membrane and cytosolic extracts of plantar skin from PTX-treated animals. b Levels of PKCε in total extract of DRG from PTX-treated animals. Animals were treated with the selective kinin B1 (DALBK; 150 nmol/kg, i.p.) or B2 (HOE 140; 50 nmol/kg, i.p.) receptor antagonists. Each group represents the mean of six animals, and the error bars indicate the SEM. *P < 0.05, significantly different from the control group. #P < 0.05, significantly different from the vehicle-treated group. One-way ANOVA followed by the Newman–Keuls post test

PKCε Mediates Hypotonicity-Induced Overt Nociception in PTX-Treated Mice

The injection of the selective PKCε activator ψεRACK (0.1 μg/paw) 5 min prior to hypotonic solution caused a significant decrease in the mechanical withdrawal threshold in comparison with that observed for saline pre-injected mice (Fig. 7a). We also assessed the involvement of PKCε in the hypotonicity-induced overt nociception in PTX-treated mice. Administration of the selective PKCε inhibitor εV1–2 (9 μg/paw) markedly prevented the hypotonicity-induced overt nociception in neuropathic animals (Fig. 7b).
Fig. 7

Activation of PKCε sensitizes paclitaxel-treated mice to hypotonicity. a Effect of a sub-effective dose of the selective PKCε activator ΨεRACK (0.1 μg/paw) on the plantar mechanical threshold after the injection of hypotonic solution (Hypo; 10 μl of deionized water per paw). b Effect of selective PKCε inhibitor εV1–2 (9 μg/paw) on the overt nociception induced by hypotonic solution (20 μl of deionized water per paw) in PTX-treated animals. Each group represents the mean of 8 (a) or 10 (b) animals, and the error bars indicate the SEM. *P < 0.05, significantly different from saline-injected (a) or control (b) groups. One-way ANOVA followed by the Newman–Keuls post test

Discussion

Paclitaxel is one of the most effective and commonly used anti-neoplastic drug. However, its major dose-limiting side effect is the development of peripheral sensory neuropathy, which remains without satisfactory treatment and compromises patient’s quality of life [2, 31]. Hence, understanding the mechanisms underlying this syndrome is critical to the discovery of new molecular targets and to the development of more effective analgesic drugs. We have recently reported that in the absence of kinin receptor (B1R and B2R) activation, there is a decrease in PTX-induced mechanical hyperalgesia [3]. Similarly, the mechanical hyperalgesia induced by PTX was shown to be reduced in TRPV4-deficient mice [21] and in animals treated with TRPV4 antisense oligodeoxynucleotides [20] or selective TRPV4 antagonists [22, 23]. Here, we show that TRPV4 channels mediate the mechanical hyperalgesia induced by the activation of kinin receptors and that this phenomenon may contribute to PTX-induced peripheral neuropathy in mice.

Although it is well known that BK causes hypersensitivity to mechanical stimuli, the mechanisms underlying this response are not completely understood [32]. Importantly, TRPV4 is a mechanotransducer channel that contributes to the mechanical hyperalgesia induced by pro-inflammatory mediators such as prostaglandins and proteases [18, 19]. Here, we investigated whether TRPV4 mediates the mechanical hyperalgesia induced by BK (a B2R agonist) and DABK (a B1R agonist) in mice. We showed that the treatment with the selective TRPV4 antagonist HC-067047 significantly inhibits the nociceptive responses elicited by both BK and DABK, suggesting that TRPV4 sensitization occurs downstream to B2R and B1R activation in order to induce mechanical hypersensitivity in mice. Since the i.pl. injection of DABK is not capable of producing any detectable overt nociceptive response [11] or mechanical hyperalgesia in mice (data not shown), DABK was given i.t. Indeed, this approach is known to produce increased sensitivity to both thermal and mechanical stimuli in mice [33, 34]. In fact, B1 receptors are reported to be constitutively and functionally expressed in the central nervous system [6, 33, 34, 35]. To our knowledge, we present here the first evidences on that kinin receptors and TRPV4 interact in vivo.

Corroborating the above data, a sub-effective dose of BK sensitized mice to the TRPV4 activator 4α-PDD or hypotonic solution and caused mechanical hyperalgesia. Nonetheless, the pre-injection of DABK (20 nmol/paw) was not able to sensitize mice to hypotonicity (data not shown). Antagonism of B2R or TRPV4, but not B1R, significantly attenuated the mechanical hyperalgesia induced by the combination of BK and hypotonicity, providing, therefore, additional evidence on the involvement of these receptors in this response. Conversely, the antagonism of B1R was not effective, possibly because BK acts preferentially on B2R and needs to be degraded in DABK in order to activate B1R [4]. Additionally, B1R is suggested to be non-functional in peripheral tissues under normal conditions and needs to be stimulated in order to mediate nociception [11, 36].

Our findings also indicate that the sensitizing effect of BK to hypotonicity is dependent on PLC and PKC activation as inhibition of their activity significantly attenuated the mechanical hyperalgesia induced by hypotonicity in BK-injected animals. The PLC/PKC signalling pathway was previously suggested to be involved in the sensitization of TRPV4 by inflammatory mediators in HEK293 cells [15]. Nonetheless, BK can also sensitize nociceptors by mechanisms other than the PLC/PKC pathway, including indirect sensitization by releasing of prostaglandins or sympathomimetic amines [37], increases in the intracellular Ca2+ concentration by opening of TRPV1 channels [12, 13] or activation of PLA2 or PKA enzymes [14, 38]. However, the sensitizing effect of BK to hypotonicity seems to be independent of these mechanisms as the blockade of such pathways did not interfere with the mechanical hyperalgesia induced by hypotonicity in BK-injected mice. Therefore, we can conclude that the sensitizing effect of BK to hypotonicity is mainly dependent on PLC and PKC activation.

It was previously demonstrated that TRPV4 and kinin receptors play a crucial role in the peripheral neuropathy induced by PTX in rodents [3, 20]. Indeed, the deletion of TRPV4 or the treatment with TRPV4 antisense oligodeoxynucleotides [20] or selective antagonists for this channel [22, 23] inhibits PTX-induced mechanical hyperalgesia in rodents. Similar results were found in mice lacking kinin receptors and also in those treated with selective kinin B1R or B2R antagonists [3]. However, to date, the downstream pathways activated by kinin receptors to maintain PTX-induced mechanical hyperalgesia are unknown. Herein, the treatment with the selective kinin receptor antagonists, DALBK and HOE 140, presented similar efficacy to that observed for the selective TRPV4 antagonist (HC-067047) on the PTX-induced mechanical hyperalgesia in mice. These results suggest that kinin receptors modulate TRPV4 channel activity. Additionally, it is possible that these receptors may act through common anti-nociceptive pathways, as both the TRPV4 and the kinin receptor antagonists exhibited similar inhibition profiles in our model.

To investigate this hypothesis, we evaluated the effect of kinin receptor antagonists on the overt nociception induced by hypotonicity in PTX-treated mice. We found that in addition to the mechanical hyperalgesia, peripheral neuropathy induced by PTX is accompanied by increased sensitivity to hypotonicity, characterized by TRPV4-dependent overt nociception [17, 20]. We show that the treatment with either DALBK (B1R antagonist) or HOE 140 (B2R antagonist) reduces hypotonicity-induced overt nociception in PTX-treated animals, suggesting that kinins act on their receptors and sensitize TRPV4 channels, thus contributing to the peripheral neuropathy induced by PTX in mice.

PKCε activation was previously suggested to contribute to the mechanical hyperalgesia caused by PTX [39] and to the sensitization of TRP channels by inflammatory mediators [9, 40, 41, 42]. Thus, we investigated whether PKCε activation acts downstream to the B1R and B2R activation in order to sensitize TRPV4 channels in our model. PTX treatment increased the levels of PKCε but did not change the ratio between membrane and cytoplasm protein expression (data not shown), suggesting a change in PKCε expression rather than in its trafficking to the cell membrane. Importantly, treatment with the selective kinin receptor antagonists reduced the levels of PKCε in both the membrane and cytosolic extracts from the plantar skin of neuropathic mice, indicating that kinins may regulate its expression. Corroborating these findings, kinin receptor antagonism reduced the levels of PKCε in DRG total extracts from PTX-treated animals. Indeed, it was recently shown that PKCε mediates the pro-nociceptive response to kinins in a mouse model of muscle pain induced by formalin [43].

We also reported here that the selective PKCε activator (ψεRACK) is able to sensitize mice to hypotonicity and, most importantly, that the treatment with the selective PKCε inhibitor εV1–2 reduces hypotonicity-induced overt nociception in PTX-treated mice in a similar manner to that observed for the kinin receptor antagonists. Collectively, our findings suggest that PKCε acts downstream to B1R and B2R activation to sensitize TRPV4 channels in PTX-treated mice. Indeed, PKCε activation was implicated in the sensitization of TRPV4 by inflammatory mediators, contributing to mechanical hyperalgesia [18]. Additionally, Chen and collaborators [22] proposed that PKCε modulates TRPV4 activity in PTX-induced peripheral neuropathy in mice, based on the similar efficacy observed for a PKCε inhibitor and a TRPV4 antagonist in mechanical hyperalgesia.

Overall, we demonstrate that kinins sensitize TRPV4 to cause mechanical hyperalgesia and that this phenomenon may contribute to PTX-induced peripheral neuropathy.

Conclusions

In summary, this study suggests that kinins sensitize TRPV4 channels to induce mechanical hyperalgesia in mice. Our results also demonstrate that this mechanism of interaction seems to contribute to the maintenance of mechanical hyperalgesia induced by PTX through the activation of PKCε. These evidences are novel and support the notion that these receptors are potential pharmacological targets for the development of new and effective analgesics to treat chronic pain.

Notes

Acknowledgments

The study was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina (FAPESC) e Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA). M.A.B and M.N.M. are PhD students funded by the CNPq. G.C.S. and F.C.D are master’s students funded by the CAPES.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Polomano RC, Bennett GJ (2001) Chemotherapy-evoked painful peripheral neuropathy. Pain Med 2(1):8–14. doi:10.1046/j.1526-4637.2001.002001008.x CrossRefPubMedGoogle Scholar
  2. 2.
    Dougherty PM, Cata JP, Cordella JV, Burton A, Weng HR (2004) Taxol-induced sensory disturbance is characterized by preferential impairment of myelinated fiber function in cancer patients. Pain 109(1–2):132–142. doi:10.1016/j.pain.2004.01.021 CrossRefPubMedGoogle Scholar
  3. 3.
    Costa R, Motta EM, Dutra RC, Manjavachi MN, Bento AF, Malinsky FR, Pesquero JB, Calixto JB (2011) Anti-nociceptive effect of kinin B(1) and B(2) receptor antagonists on peripheral neuropathy induced by paclitaxel in mice. Br J Pharmacol 164(2b):681–693. doi:10.1111/j.1476-5381.2011.01408.x CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Calixto JB, Medeiros R, Fernandes ES, Ferreira J, Cabrini DA, Campos MM (2004) Kinin B1 receptors: key G-protein-coupled receptors and their role in inflammatory and painful processes. Br J Pharmacol 143(7):803–818. doi:10.1038/sj.bjp.0706012 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Marceau F, Regoli D (2004) Bradykinin receptor ligands: therapeutic perspectives. Nat Rev Drug Discov 3(10):845–852. doi:10.1038/nrd1522 CrossRefPubMedGoogle Scholar
  6. 6.
    Ma QP, Heavens R (2001) Basal expression of bradykinin B(1) receptor in the spinal cord in humans and rats. Neuroreport 12(11):2311–2314CrossRefPubMedGoogle Scholar
  7. 7.
    Ma QP (2001) The expression of bradykinin B(1) receptors on primary sensory neurones that give rise to small caliber sciatic nerve fibres in rats. Neuroscience 107(4):665–673CrossRefPubMedGoogle Scholar
  8. 8.
    Cesare P, McNaughton P (1996) A novel heat-activated current in nociceptive neurons and its sensitization by bradykinin. Proc Natl Acad Sci U S A 93(26):15435–15439CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Cesare P, Dekker LV, Sardini A, Parker PJ, McNaughton PA (1999) Specific involvement of PKC-epsilon in sensitization of the neuronal response to painful heat. Neuron 23(3):617–624CrossRefPubMedGoogle Scholar
  10. 10.
    Chuang HH, Prescott ED, Kong H, Shields S, Jordt SE, Basbaum AI, Chao MV, Julius D (2001) Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition. Nature 411(6840):957–962. doi:10.1038/35082088 CrossRefPubMedGoogle Scholar
  11. 11.
    Ferreira J, da Silva GL, Calixto JB (2004) Contribution of vanilloid receptors to the overt nociception induced by B2 kinin receptor activation in mice. Br J Pharmacol 141(5):787–794. doi:10.1038/sj.bjp.0705546 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Bandell M, Story GM, Hwang SW, Viswanath V, Eid SR, Petrus MJ, Earley TJ, Patapoutian A (2004) Noxious cold ion channel TRPA1 is activated by pungent compounds and bradykinin. Neuron 41(6):849–857CrossRefPubMedGoogle Scholar
  13. 13.
    Bautista DM, Jordt SE, Nikai T, Tsuruda PR, Read AJ, Poblete J, Yamoah EN, Basbaum AI et al (2006) TRPA1 mediates the inflammatory actions of environmental irritants and proalgesic agents. Cell 124(6):1269–1282. doi:10.1016/j.cell.2006.02.023 CrossRefPubMedGoogle Scholar
  14. 14.
    Wang S, Dai Y, Fukuoka T, Yamanaka H, Kobayashi K, Obata K, Cui X, Tominaga M et al (2008) Phospholipase C and protein kinase A mediate bradykinin sensitization of TRPA1: a molecular mechanism of inflammatory pain. Brain 131(Pt 5):1241–1251. doi:10.1093/brain/awn060 PubMedGoogle Scholar
  15. 15.
    Fan HC, Zhang X, McNaughton PA (2009) Activation of the TRPV4 ion channel is enhanced by phosphorylation. J Biol Chem 284(41):27884–27891. doi:10.1074/jbc.M109.028803 CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Liedtke W, Tobin DM, Bargmann CI, Friedman JM (2003) Mammalian TRPV4 (VR-OAC) directs behavioral responses to osmotic and mechanical stimuli in Caenorhabditis elegans. Proc Natl Acad Sci U S A 100(Suppl 2):14531–14536. doi:10.1073/pnas.2235619100 CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Alessandri-Haber N, Yeh JJ, Boyd AE, Parada CA, Chen X, Reichling DB, Levine JD (2003) Hypotonicity induces TRPV4-mediated nociception in rat. Neuron 39(3):497–511CrossRefPubMedGoogle Scholar
  18. 18.
    Alessandri-Haber N, Dina OA, Joseph EK, Reichling D, Levine JD (2006) A transient receptor potential vanilloid 4-dependent mechanism of hyperalgesia is engaged by concerted action of inflammatory mediators. The Journal of Neuroscience 26(14):3864–3874. doi:10.1523/JNEUROSCI.5385-05.2006 CrossRefPubMedGoogle Scholar
  19. 19.
    Grant AD, Cottrell GS, Amadesi S, Trevisani M, Nicoletti P, Materazzi S, Altier C, Cenac N et al (2007) Protease-activated receptor 2 sensitizes the transient receptor potential vanilloid 4 ion channel to cause mechanical hyperalgesia in mice. J Physiol 578(Pt 3):715–733. doi:10.1113/jphysiol.2006.121111 CrossRefPubMedGoogle Scholar
  20. 20.
    Alessandri-Haber N, Dina OA, Yeh JJ, Parada CA, Reichling DB, Levine JD (2004) Transient receptor potential vanilloid 4 is essential in chemotherapy-induced neuropathic pain in the rat. The Journal of Neuroscience 24(18):4444–4452. doi:10.1523/JNEUROSCI.0242-04.2004 CrossRefPubMedGoogle Scholar
  21. 21.
    Alessandri-Haber N, Dina OA, Joseph EK, Reichling DB, Levine JD (2008) Interaction of transient receptor potential vanilloid 4, integrin, and SRC tyrosine kinase in mechanical hyperalgesia. The Journal of Neuroscience 28(5):1046–1057. doi:10.1523/JNEUROSCI.4497-07.2008 CrossRefPubMedGoogle Scholar
  22. 22.
    Chen Y, Yang C, Wang ZJ (2011) Proteinase-activated receptor 2 sensitizes transient receptor potential vanilloid 1, transient receptor potential vanilloid 4, and transient receptor potential ankyrin 1 in paclitaxel-induced neuropathic pain. Neuroscience 193:440–451. doi:10.1016/j.neuroscience.2011.06.085 CrossRefPubMedGoogle Scholar
  23. 23.
    Materazzi S, Fusi C, Benemei S, Pedretti P, Patacchini R, Nilius B, Prenen J, Creminon C et al (2012) TRPA1 and TRPV4 mediate paclitaxel-induced peripheral neuropathy in mice via a glutathione-sensitive mechanism. Pflugers Archiv 463(4):561–569. doi:10.1007/s00424-011-1071-x CrossRefPubMedGoogle Scholar
  24. 24.
    Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG, Group NCRRGW (2010) Animal research: reporting in vivo experiments: the ARRIVE guidelines. Br J Pharmacol 160(7):1577–1579. doi:10.1111/j.1476-5381.2010.00872.x CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Hylden JL, Wilcox GL (1980) Intrathecal morphine in mice: a new technique. Eur J Pharmacol 67(2–3):313–316CrossRefPubMedGoogle Scholar
  26. 26.
    Costa R, Manjavachi MN, Motta EM, Marotta DM, Juliano L, Torres HA, Pesquero JB, Calixto JB (2010) The role of kinin B1 and B2 receptors in the scratching behaviour induced by proteinase-activated receptor-2 agonists in mice. Br J Pharmacol 159(4):888–897. doi:10.1111/j.1476-5381.2009.00571.x CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Costa R, Motta EM, Manjavachi MN, Cola M, Calixto JB (2012) Activation of the alpha-7 nicotinic acetylcholine receptor (alpha7 nAchR) reverses referred mechanical hyperalgesia induced by colonic inflammation in mice. Neuropharmacology 63(5):798–805. doi:10.1016/j.neuropharm.2012.06.004 CrossRefPubMedGoogle Scholar
  28. 28.
    Dixon WJ (1980) Efficient analysis of experimental observations. Annu Rev Pharmacol Toxicol 20:441–462. doi:10.1146/annurev.pa.20.040180.002301 CrossRefPubMedGoogle Scholar
  29. 29.
    Dutra RC, Bicca MA, Segat GC, Silva KA, Motta EM, Pianowski LF, Costa R, Calixto JB (2015) The antinociceptive effects of the tetracyclic triterpene euphol in inflammatory and neuropathic pain models: the potential role of PKCepsilon. Neuroscience 303:126–137. doi:10.1016/j.neuroscience.2015.06.051 CrossRefPubMedGoogle Scholar
  30. 30.
    Everaerts W, Zhen X, Ghosh D, Vriens J, Gevaert T, Gilbert JP, Hayward NJ, McNamara CR et al (2010) Inhibition of the cation channel TRPV4 improves bladder function in mice and rats with cyclophosphamide-induced cystitis. Proc Natl Acad Sci U S A 107(44):19084–19089. doi:10.1073/pnas.1005333107 CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Rivera E, Cianfrocca M (2015) Overview of neuropathy associated with taxanes for the treatment of metastatic breast cancer. Cancer Chemother Pharmacol 75(4):659–670. doi:10.1007/s00280-014-2607-5 CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Mizumura K, Sugiura T, Katanosaka K, Banik RK, Kozaki Y (2009) Excitation and sensitization of nociceptors by bradykinin: what do we know? Exp Brain Res 196(1):53–65. doi:10.1007/s00221-009-1814-5 CrossRefPubMedGoogle Scholar
  33. 33.
    Ferreira J, Campos MM, Araujo R, Bader M, Pesquero JB, Calixto JB (2002) The use of kinin B1 and B2 receptor knockout mice and selective antagonists to characterize the nociceptive responses caused by kinins at the spinal level. Neuropharmacology 43(7):1188–1197CrossRefPubMedGoogle Scholar
  34. 34.
    Fox A, Wotherspoon G, McNair K, Hudson L, Patel S, Gentry C, Winter J (2003) Regulation and function of spinal and peripheral neuronal B1 bradykinin receptors in inflammatory mechanical hyperalgesia. Pain 104(3):683–691CrossRefPubMedGoogle Scholar
  35. 35.
    Ma QP, Hill R, Sirinathsinghji D (2000) Basal expression of bradykinin B1 receptor in peripheral sensory ganglia in the rat. Neuroreport 11(18):4003–4005CrossRefPubMedGoogle Scholar
  36. 36.
    Ferreira J, Triches KM, Medeiros R, Cabrini DA, Mori MA, Pesquero JB, Bader M, Calixto JB (2008) The role of kinin B1 receptors in the nociception produced by peripheral protein kinase C activation in mice. Neuropharmacology 54(3):597–604. doi:10.1016/j.neuropharm.2007.11.008 CrossRefPubMedGoogle Scholar
  37. 37.
    Cunha TM, Verri WA Jr, Fukada SY, Guerrero AT, Santodomingo-Garzon T, Poole S, Parada CA, Ferreira SH et al (2007) TNF-alpha and IL-1beta mediate inflammatory hypernociception in mice triggered by B1 but not B2 kinin receptor. Eur J Pharmacol 573(1–3):221–229. doi:10.1016/j.ejphar.2007.07.007 CrossRefPubMedGoogle Scholar
  38. 38.
    Yanaga F, Hirata M, Koga T (1991) Evidence for coupling of bradykinin receptors to a guanine-nucleotide binding protein to stimulate arachidonate liberation in the osteoblast-like cell line, MC3T3-E1. Biochim Biophys Acta 1094(2):139–146CrossRefPubMedGoogle Scholar
  39. 39.
    Dina OA, Chen X, Reichling D, Levine JD (2001) Role of protein kinase Cepsilon and protein kinase A in a model of paclitaxel-induced painful peripheral neuropathy in the rat. Neuroscience 108(3):507–515CrossRefPubMedGoogle Scholar
  40. 40.
    Aley KO, Messing RO, Mochly-Rosen D, Levine JD (2000) Chronic hypersensitivity for inflammatory nociceptor sensitization mediated by the epsilon isozyme of protein kinase C. The Journal of Neuroscience 20(12):4680–4685PubMedGoogle Scholar
  41. 41.
    Zhang H, Cang CL, Kawasaki Y, Liang LL, Zhang YQ, Ji RR, Zhao ZQ (2007) Neurokinin-1 receptor enhances TRPV1 activity in primary sensory neurons via PKCepsilon: a novel pathway for heat hyperalgesia. The Journal of Neuroscience 27(44):12067–12077. doi:10.1523/JNEUROSCI.0496-07.2007 CrossRefPubMedGoogle Scholar
  42. 42.
    Sachs D, Villarreal C, Cunha F, Parada C, Ferreira S (2009) The role of PKA and PKCepsilon pathways in prostaglandin E2-mediated hypernociception. Br J Pharmacol 156(5):826–834. doi:10.1111/j.1476-5381.2008.00093.x CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Meotti FC, Campos R, da Silva K, Paszcuk AF, Costa R, Calixto JB (2012) Inflammatory muscle pain is dependent on the activation of kinin B(1) and B(2) receptors and intracellular kinase pathways. Br J Pharmacol 166(3):1127–1139. doi:10.1111/j.1476-5381.2012.01830.x CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Robson Costa
    • 1
    • 2
  • Maíra A. Bicca
    • 1
  • Marianne N. Manjavachi
    • 1
  • Gabriela C. Segat
    • 1
  • Fabiana Chaves Dias
    • 2
  • Elizabeth S. Fernandes
    • 3
    • 4
  • João B. Calixto
    • 1
    • 5
  1. 1.Departamento de Farmacologia, Centro de Ciências BiológicasUniversidade Federal de Santa CatarinaFlorianópolisBrazil
  2. 2.Programa de Pós-graduação de Ciências Farmacêuticas, Faculdade de FarmáciaUniversidade Federal do Rio de JaneiroRJBrazil
  3. 3.Programa de Pós-Graduação em Biologia ParasitáriaUniversidade CeumaSão LuísBrazil
  4. 4.Vascular Biology and Inflammation Section, Cardiovascular DivisionKing’s College LondonLondonUK
  5. 5.Centro de Inovação e Ensaios Pré-clínicos (CIEnP)FlorianópolisBrazil

Personalised recommendations