Localisation, mécanisme d’induction et rôle physiopathologique du récepteur B1 des kinines dans de modèles expérimentaux de douleur chez le rat

Les kinines sont des peptides neuro- et vaso- actifs impliqués dans les processus hémodynamiques, inflammatoires et douloureux. Leurs effets biologiques sont produits par l’entremise de deux types de récepteurs couplés aux protéines G, soit B1 (B1R) et B2 (B2R). Le B1R est inductible, son expression est augmentée à la suite d’un dommage tissulaire ou de l’exposition à des endotoxines bactériennes (lipopolysaccharide bactérien (LPS)), à des cytokines pro-inflammatoires (interleukine-1β (IL-1β), facteur de nécrose tumorale-α (TNF-α)) ou à des espèces réactives oxygénées (ROS). Les travaux présentés dans cette thèse avaient pour objectif d’élucider et/ou de raffiner les connaissances sur 1) la localisation, 2) le mécanisme d’induction et 3) le rôle physiopathologique du B1R dans des modèles expérimentaux de douleur chez le rat. Nos données ont permis de démontrer pour la première fois que le B1R est augmenté de façon significative dans la moelle épinière du rat diabétique de type 1 où il est localisé sur les fibres sensorielles de type C, les astrocytes et les cellules de la microglie (1er article). Également, l’inhibition de l’activation des cellules de la microglie supprime les neuropathies diabétiques, l’expression de médiateurs pro-inflammatoires ainsi que l’activité pro-nociceptive du B1R (2e et 3e articles). Finalement, nous avons démontré que la stimulation systémique du TRPV1 par la capsaïcine induit une surexpression du B1R au niveau microgliale, via un mécanisme impliquant l’augmentation de la production de ROS et possiblement de cytokines (4e article). Ces données nous permettent de mieux comprendre les mécanismes impliqués dans l’expression et l’activité du B1R. Aussi, elles nous permettent d’imaginer de nouvelles stratégies pour prévenir l’induction du B1R (inhibition du TRPV1) ou son activité délétère (inhibition de l’activation des cellules de la microglie) dans la douleur inflammatoire et neuropathique. Kinins are vaso- and neuro-active peptides involved in hemodynamic, inflammatory and pain processes. Their biological effects are mediated by two G Protein Coupled Receptors (GPCR), termed B2R (constitutive) and B1R (inducible). B1R is expressed following tissue damage or exposure to bacterial endotoxin (LPS), pro-inflammatory cytokines (IL-1β, TNF-α) and increased reactive oxygen species (ROS) levels. The objectives of this doctoral thesis were to define 1) the localisation, 2) the mechanism of induction and 3) the pathophysiological role of B1R in experimental models of pain in rat. Our data showed that B1R is significantly upregulated on sensory C fibers, astrocytes and microglia in spinal cord of type 1 diabetic rat (paper #1). Moreover, pharmacological inhibition of microglia reversed diabetic pain neuropathy, reduced levels of pro-inflammatory mediators and prevented B1R pro-nociceptive activity (papers #2 and 3). Finally, our data showed that systemic stimulation of TRPV1 with capsaicin upregulated B1R expression, mainly on microglia, through the increase of ROS and possibly cytokines (paper #4). Altogether, these data increased our knowledge related to B1R mechanism of induction and B1R activity. Also, these data shed light on new strategies to prevent B1R expression (TRPV1 blockade) and B1R deleterious activity (inhibition of microglia activation) in inflammatory and neuropathic pain

Key role for spinal dorsal horn microglial kinin B1 receptor in early diabetic pain neuropathy

<p>Abstract</p> <p>Background</p> <p>The pro-nociceptive kinin B₁receptor (B₁R) is upregulated on sensory C-fibres, astrocytes and microglia in the spinal cord of streptozotocin (STZ)-diabetic rat. This study aims at defining the role of microglial kinin B₁R in diabetic pain neuropathy.</p> <p>Methods</p> <p>Sprague-Dawley rats were made diabetic with STZ (65 mg/kg, i.p.), and 4 days later, two specific inhibitors of microglial cells (fluorocitrate, 1 nmol, i.t.; minocycline, 10 mg/kg, i.p.) were administered to assess the impact on thermal hyperalgesia, allodynia and mRNA expression (qRT-PCR) of B₁R and pro-inflammatory markers. Spinal B₁R binding sites ((¹²⁵I)-HPP-desArg¹⁰-Hoe 140) were also measured by quantitative autoradiography. Inhibition of microglia was confirmed by confocal microscopy with the specific marker Iba-1. Effects of intrathecal and/or systemic administration of B₁R agonist (des-Arg⁹-BK) and antagonists (SSR240612 and R-715) were measured on neuropathic pain manifestations.</p> <p>Results</p> <p>STZ-diabetic rats displayed significant tactile and cold allodynia compared with control rats. Intrathecal or peripheral blockade of B₁R or inhibition of microglia reversed time-dependently tactile and cold allodynia in diabetic rats without affecting basal values in control rats. Microglia inhibition also abolished thermal hyperalgesia and the enhanced allodynia induced by intrathecal des-Arg⁹-BK without affecting hyperglycemia in STZ rats. The enhanced mRNA expression (B₁R, IL-1β, TNF-α, TRPV1) and Iba-1 immunoreactivity in the STZ spinal cord were normalized by fluorocitrate or minocycline, yet B₁R binding sites were reduced by 38%.</p> <p>Conclusion</p> <p>The upregulation of kinin B₁R in spinal dorsal horn microglia by pro-inflammatory cytokines is proposed as a crucial mechanism in early pain neuropathy in STZ-diabetic rats.</p

Sense and Immunity: Context-Dependent Neuro-Immune Interplay

The sensory nervous and immune systems, historically considered autonomous, actually work in concert to promote host defense and tissue homeostasis. These systems interact with each other through a common language of cell surface G protein-coupled receptors and receptor tyrosine kinases as well as cytokines, growth factors, and neuropeptides. While this bidirectional communication is adaptive in many settings, helping protect from danger, it can also become maladaptive and contribute to disease pathophysiology. The fundamental logic of how, where, and when sensory neurons and immune cells contribute to either health or disease remains, however, unclear. Our lab and others’ have begun to explore how this neuro-immune reciprocal dialog contributes to physiological and pathological immune responses and sensory disorders. The cumulative results collected so far indicate that there is an important role for nociceptors (noxious stimulus detecting sensory neurons) in driving immune responses, but that this is highly context dependent. To illustrate this concept, we present our findings in a model of airway inflammation, in which nociceptors seem to have major involvement in type 2 but not type 1 adaptive immunity

Neurons and Microglia; A Sickly-Sweet Duo in Diabetic Pain Neuropathy

Diabetes is a common condition characterized by persistent hyperglycemia. High blood sugar primarily affects cells that have a limited capacity to regulate their glucose intake. These cells include capillary endothelial cells in the retina, mesangial cells in the renal glomerulus, Schwann cells, and neurons of the peripheral and central nervous systems. As a result, hyperglycemia leads to largely intractable complications such as retinopathy, nephropathy, hypertension, and neuropathy. Diabetic pain neuropathy is a complex and multifactorial disease that has been associated with poor glycemic control, longer diabetes duration, hypertension, advanced age, smoking status, hypoinsulinemia, and dyslipidemia. While many of the driving factors involved in diabetic pain are still being investigated, they can be broadly classified as either neuron -intrinsic or -extrinsic. In neurons, hyperglycemia impairs the polyol pathway, leading to an overproduction of reactive oxygen species and reactive nitrogen species, an enhanced formation of advanced glycation end products, and a disruption in Na+/K+ ATPase pump function. In terms of the extrinsic pathway, hyperglycemia leads to the generation of both overactive microglia and microangiopathy. The former incites a feed-forward inflammatory loop that hypersensitizes nociceptor neurons, as observed at the onset of diabetic pain neuropathy. The latter reduces neurons' access to oxygen, glucose and nutrients, prompting reductions in nociceptor terminal expression and losses in sensation, as observed in the later stages of diabetic pain neuropathy. Overall, microglia can be seen as potent and long-lasting amplifiers of nociceptor neuron activity, and may therefore constitute a potential therapeutic target in the treatment of diabetic pain neuropathy

Impact of Climate Change on the Relict Tropical Fish Fauna of Central Sahara: Threat for the Survival of Adrar Mountains Fishes, Mauritania

Background: Four central Sahara mountainous massifs provide habitats for relict populations of fish. In the Adrar of Mauritania all available data on the presence and distribution of fish come from pre-1960 surveys where five fish species were reported: Barbus pobeguini, Barbus macrops, Barbus mirei, Sarotherodon galilaeus, and Clarias anguillaris. Since 1970, drought has had a severe impact in the Adrar where rainfall decreased by 35%. To investigate whether the relict populations of fish have survived the continuing drought, a study was carried out from 2004 to 2008. Methodology/Principal Findings: An inventory of perennial bodies of water was drawn up using a literature review and analysis of topographical and hydrological maps. Field surveys were carried out in order to locate the bodies of water described in the literature, identify the presence of fish, determine which species were present and estimate their abundance. The thirteen sites where the presence of fish was observed in the 1950s -Ksar Torchane, Ilij, Molomhar, Agueni, Tachot, Hamdoun, Terjit, Toungad, El Berbera, Timagazine, Dayet el Mbarek, Dayet et-Tefla, Nkedei-were located and surveyed. The Ksar Torchane spring -type locality and the only known locality of B. mirei- has dried up at the height of the drought in 1984, and any fish populations have since become extinct there. The Timagazine, Dayet el Mbarek and Dayet et-Tefla pools have become ephemeral. The Hamdoun guelta appears to be highly endangered. The fish populations at the other sites remain unchanged. Four perennial pools which are home to populations of B. pobeguini are newly recorded. Conclusion/Significance: The tropical relict fish populations of the Adrar mountains of Mauritania appear to be highly endangered. Of thirteen previously recorded populations, four have become extinct since the beginning of the drought period. New fish population extinctions may occur should low levels of annual rainfall be repeated

A value-based comparison of the management of ambulatory respiratory diseases in walk-in clinics, primary care practices, and emergency departments : protocol for a multicenter prospective cohort study

Background: In Canada, 30%-60% of patients presenting to emergency departments are ambulatory. This category has been labeled as a source of emergency department overuse. Acting on the presumption that primary care practices and walk-in clinics offer equivalent care at a lower cost, governments have invested massively in improving access to these alternative settings in the hope that patients would present there instead when possible, thereby reducing the load on emergency departments. Data in support of this approach remain scarce and equivocal. Objective: The aim of this study is to compare the value of care received in emergency departments, walk-in clinics, and primary care practices by ambulatory patients with upper respiratory tract infection, sinusitis, otitis media, tonsillitis, pharyngitis, bronchitis, influenza-like illness, pneumonia, acute asthma, or acute exacerbation of chronic obstructive pulmonary disease. Methods: A multicenter prospective cohort study will be performed in Ontario and Québec. In phase 1, a time-driven activity-based costing method will be applied at each of the 15 study sites. This method uses time as a cost driver to allocate direct costs (eg, medication), consumable expenditures (eg, needles), overhead costs (eg, building maintenance), and physician charges to patient care. Thus, the cost of a care episode will be proportional to the time spent receiving the care. At the end of this phase, a list of care process costs will be generated and used to calculate the cost of each consultation during phase 2, in which a prospective cohort of patients will be monitored to compare the care received in each setting. Patients aged 18 years and older, ambulatory throughout the care episode, and discharged to home with one of the aforementioned targeted diagnoses will be considered. The estimated sample size is 1485 patients. The 3 types of care settings will be compared on the basis of primary outcomes in terms of the proportion of return visits to any site 3 and 7 days after the initial visit and the mean cost of care. The secondary outcomes measured will include scores on patient-reported outcome and experience measures and mean costs borne wholly by patients. We will use multilevel generalized linear models to compare the care settings and an overlap weights approach to adjust for confounding factors related to age, sex, gender, ethnicity, comorbidities, registration with a family physician, socioeconomic status, and severity of illness. Results: Phase 1 will begin in 2021 and phase 2, in 2023. The results will be available in 2025. Conclusions: The end point of our program will be for deciders, patients, and care providers to be able to determine the most appropriate care setting for the management of ambulatory emergency respiratory conditions, based on the quality and cost of care associated with each alternative

Bulk organic geochemistry of sediments from Puyehue Lake and its watershed (Chile, 40°S) : implications for paleoenvironmental reconstructions

Author Posting. © Elsevier B.V., 2009. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Palaeogeography, Palaeoclimatology, Palaeoecology 294 (2010): 56-71, doi:10.1016/j.palaeo.2009.03.012.Since the last deglaciation, the mid-latitudes of the southern Hemisphere have undergone considerable environmental changes. In order to better understand the response of continental ecosystems to paleoclimate changes in southern South America, we investigated the sedimentary record of Puyehue Lake, located in the western piedmont of the Andes in south-central Chile (40°S). We analyzed the elemental (C, N) and stable isotopic (δ13C, δ15N) composition of the sedimentary organic matter preserved in the lake and its watershed to estimate the relative changes in the sources of sedimentary organic carbon through space and time. The geochemical signature of the aquatic and terrestrial end-members was determined on samples of lake particulate organic matter (N/C: 0.130) and Holocene paleosols (N/C: 0.069), respectively. A simple mixing equation based on the N/C ratio of these end-members was then used to estimate the fraction of terrestrial carbon (ƒT) preserved in the lake sediments. Our approach was validated using surface sediment samples, which show a strong relation between ƒT and distance to the main rivers and to the shore. We further applied this equation to an 11.22 m long sediment core to reconstruct paleoenvironmental changes in Puyehue Lake and its watershed during the last 17.9 kyr. Our data provide evidence for a first warming pulse at 17.3 cal kyr BP, which triggered a rapid increase in lake diatom productivity, lagging the start of a similar increase in sea surface temperature (SST) off Chile by 1500 years. This delay is best explained by the presence of a large glacier in the lake watershed, which delayed the response time of the terrestrial proxies and limited the concomitant expansion of the vegetation in the lake watershed (low ƒT). A second warming pulse at 12.8 cal kyr BP is inferred from an increase in lake productivity and a major expansion of the vegetation in the lake watershed, demonstrating that the Puyehue glacier had considerably retreated from the watershed. This second warming pulse is synchronous with a 2°C increase in SST off the coast of Chile, and its timing corresponds to the beginning of the Younger Dryas Chronozone. These results contribute to the mounting evidence that the climate in the mid-latitudes of the southern Hemisphere was warming during the Younger Dryas Chronozone, in agreement with the bipolar see-saw hypothesis.This research was partly supported by the Belgian OSTC project EV/12/10B "A continuous Holocene record of ENSO variability in southern Chile". S.B. is supported by a BAEF fellowship (Belgian American Educational Foundation), and by an EU Marie Curie Outgoing Fellowship under the FP6 programme

Silencing Nociceptor Neurons Reduces Allergic Airway Inflammation

Lung nociceptors initiate cough and bronchoconstriction. To elucidate if these fibers also contribute to allergic airway inflammation, we stimulated lung nociceptors with capsaicin and observed increased neuropeptide release and immune cell infiltration. In contrast, ablating Nav1.8(+) sensory neurons or silencing them with QX-314, a charged sodium channel inhibitor that enters via large-pore ion channels to specifically block nociceptors, substantially reduced ovalbumin- or house-dust-mite-induced airway inflammation and bronchial hyperresponsiveness. We also discovered that IL-5, a cytokine produced by activated immune cells, acts directly on nociceptors to induce the release of vasoactive intestinal peptide (VIP). VIP then stimulates CD4(+) and resident innate lymphoid type 2 cells, creating an inflammatory signaling loop that promotes allergic inflammation. Our results indicate that nociceptors amplify pathological adaptive immune responses and that silencing these neurons with QX-314 interrupts this neuro-immune interplay, revealing a potential new therapeutic strategy for asthma