RESUMEN
Host proteins released by the activated endothelial cells during SARS-CoV-2 infection are implicated to be involved in coagulation and endothelial dysfunction. However, the underlying mechanism that governs the vascular dysfunction and disease severity in COVID-19 remains obscure. The study evaluated the serum levels of Bradykinin, Kallikrein, SERPIN A, and IL-18 in COVID-19 (N-42 with 20 moderate and 22 severe) patients compared to healthy controls (HC: N-10) using ELISA at the day of admission (DOA) and day 7 post-admission. The efficacy of the protein levels in predicting disease severity was further determined using machine learning models. The levels of bradykinins and SERPIN A were higher (P ≤ 0.001) in both severe and moderate cases on day 7 post-admission compared to DOA. All the soluble proteins studied were found to elevated (P ≤ 0.01) in severe compared to moderate in day 7 and were positively correlated (P ≤ 0.001) with D-dimer, a marker for coagulation. ROC analysis identified that SERPIN A, IL-18, and bradykinin could predict the clinical condition of COVID-19 with AUC values of 1, 0.979, and 1, respectively. Among the models trained using univariate model analysis, SERPIN A emerged as a strong prognostic biomarker for COVID-19 disease severity. The serum levels of SERPIN A in conjunction with the coagulation marker D-dimer, serve as a predictive indicator for COVID-19 clinical outcomes. However, studies are required to ascertain the role of these markers in disease virulence.
Asunto(s)
Biomarcadores , Bradiquinina , COVID-19 , Interleucina-18 , SARS-CoV-2 , Humanos , COVID-19/sangre , COVID-19/diagnóstico , Biomarcadores/sangre , Femenino , Masculino , Persona de Mediana Edad , Pronóstico , Interleucina-18/sangre , Bradiquinina/sangre , Adulto , Anciano , Productos de Degradación de Fibrina-Fibrinógeno/metabolismo , Productos de Degradación de Fibrina-Fibrinógeno/análisis , Índice de Severidad de la Enfermedad , Endotelio Vascular/metabolismo , Calicreínas/sangre , alfa 1-Antitripsina/sangreRESUMEN
Angioedema is a prevailing symptom in different diseases, frequently occurring in the presence of urticaria. Recurrent angioedema without urticaria (AE) can be hereditary (HAE) and acquired (AAE), and several subtypes can be distinguished, although clinical presentation is quite similar in some of them. They present with subcutaneous and mucosal swellings, affecting extremities, face, genitals, bowels, and upper airways. AE is commonly misdiagnosed due to restricted access and availability of appropriate laboratorial tests. HAE with C1 inhibitor defect is associated with quantitative and/or functional deficiency. Although bradykinin-mediated disease results mainly from disturbance in the kallikrein-kinin system, traditionally complement evaluation has been used for diagnosis. Diagnosis is established by nephelometry, turbidimetry, or radial immunodiffusion for quantitative measurement of C1 inhibitor, and chromogenic assay or ELISA has been used for functional C1-INH analysis. Wrong handling of the samples can lead to misdiagnosis and, consequently, mistaken inappropriate approaches. Dried blood spot (DBS) tests have been used for decades in newborn screening for certain metabolic diseases, and there has been growing interest in their use for other congenital conditions. Recently, DBS is now proposed as an efficient tool to diagnose HAE with C1 inhibitor deficiency, and its use would improve the access to outbound areas and family members. Regarding HAE with normal C1 inhibitor, complement assays' results are normal and the genetic sequencing of target genes, such as exon 9 of F12 and PLG, is the only available method. New methods to measure cleaved high-molecular-weight kininogen and activated plasma kallikrein have emerged as potential biochemical tests to identify bradykinin-mediated angioedema. Validated biomarkers of kallikrein-kinin system activation could be helpful in differentiating mechanisms of angioedema. Our aim is to focus on the capability to differentiate histaminergic AE from bradykinin-mediated AE. In addition, we will describe the challenges developing specific tests like direct bradykinin measurements. The need for quality tests to improve the diagnosis is well represented by the variability of results in functional assays.
Asunto(s)
Angioedema/diagnóstico , Angioedemas Hereditarios/diagnóstico , Errores Diagnósticos/prevención & control , Angioedema/sangre , Angioedema/inmunología , Angioedemas Hereditarios/sangre , Angioedemas Hereditarios/genética , Angioedemas Hereditarios/inmunología , Biomarcadores/sangre , Biomarcadores/metabolismo , Bradiquinina/sangre , Bradiquinina/inmunología , Bradiquinina/metabolismo , Proteína Inhibidora del Complemento C1/análisis , Proteína Inhibidora del Complemento C1/genética , Proteína Inhibidora del Complemento C1/metabolismo , Análisis Mutacional de ADN , Diagnóstico Diferencial , Pruebas con Sangre Seca/métodos , Ensayo de Inmunoadsorción Enzimática , Factor XII/genética , Humanos , Mutación , Plasminógeno/genética , RecurrenciaRESUMEN
The renin-angiotensin-aldosterone system and its metabolites play an important role in homeostasis of body, especially the cardiovascular system. In this study, we discuss the imbalance of multiple systems during the infection and the importance of therapeutic choice, dosing, and laboratory monitoring of cardiac and anti-coagulant therapies in COVID-19 patients. The crosstalk between angiotensin, kinin-kallikrein system, as well as inflammatory and coagulation systems plays an essential role in COVID-19. Cardiac complications and coagulopathies imply the crosstalks between the mentioned systems. We believe that the blockage of bradykinin can be a good option in the management of COVID-19 and CVD in patients and that supportive treatment of respiratory and cardiologic complications is needed in COVID-19 patients. Ninety-one percent of COVID-19 patients who were admitted to hospital with a prolonged aPTT were positive for lupus anticoagulant, which increases the risk of thrombosis and prolonged aPTT. Therefore, the question that is posed at this juncture is whether it is safe to use the prophylactic dose of heparin particularly in those with elevated D-dimer levels. It should be noted that timing is of high importance in anti-coagulant therapy; therefore, we should consider the level of D-dimer, fibrinogen, drug-drug interactions, and risk factors during thromboprophylaxis administration. Fibrinogen is an independent predictor of resistance to heparin and should be considered before thromboprophylaxis. Alteplase and Futhan might be a good choice to assess the condition of heparin resistance. Finally, the treatment option, dosing, and laboratory monitoring of anticoagulant therapy are critical decisions in COVID-19 patients.
Asunto(s)
COVID-19 , SARS-CoV-2 , Trombosis , Anticoagulantes/administración & dosificación , Anticoagulantes/uso terapéutico , Bradiquinina/sangre , COVID-19/complicaciones , COVID-19/inmunología , COVID-19/fisiopatología , COVID-19/terapia , Síndrome de Liberación de Citoquinas/inmunología , Síndrome de Liberación de Citoquinas/virología , Productos de Degradación de Fibrina-Fibrinógeno/análisis , Humanos , Inflamación/inmunología , Inflamación/virología , Calicreínas/sangre , Sistema Renina-Angiotensina/inmunología , Sistema Renina-Angiotensina/fisiología , Trombosis/tratamiento farmacológico , Trombosis/prevención & control , Trombosis/virologíaRESUMEN
The kallikrein-kinin system (KKS) is involved in many physiological and pathophysiological processes and is assumed to be connected to the development of clinical symptoms of angioedema or COVID-19, among other diseases. However, despite its diverse role in the regulation of physiological and pathophysiological functions, knowledge about the KKS in vivo remains limited. The short half-lives of kinins, their low abundance and structural similarities and the artificial generation of the kinin bradykinin greatly hinder reliable and accurate determination of kinin levels in plasma. To address these issues, a sensitive LC-MS/MS platform for the comprehensive and simultaneous determination of the four active kinins bradykinin, kallidin, des-Arg(9)-bradykinin and des-Arg(10)-kallidin and their major metabolites bradykinin 2-9, bradykinin 1-7 and bradykinin 1-5 was developed. This platform was validated according to the bioanalytical guideline of the US Food and Drug Administration regarding linearity, accuracy, precision, sensitivity, carry-over, recovery, parallelism, matrix effects and stability in plasma of healthy volunteers. The validated platform encompassed a broad calibration curve range from 2.0-15.3 pg/mL (depending on the kinin) up to 1000 pg/mL, covering the expected concentrations in disease states. No source-dependent matrix effects were identified, and suitable stability of the analytes in plasma was observed. The applicability of the developed platform was proven by the determination of endogenous levels in healthy volunteers, whose plasma kinin levels were successfully detected in the low pg/mL range. The established platform facilitates the investigation of kinin-mediated diseases (e.g. angioedema, COVID-19) and enables the assessment of the impact of altered enzyme activities on the formation or degradation of kinins.
Asunto(s)
Bradiquinina/análogos & derivados , Bradiquinina/sangre , Calidina/análogos & derivados , Calidina/sangre , Sistema Calicreína-Quinina , Espectrometría de Masas en Tándem/métodos , COVID-19/sangre , Cromatografía Liquida/métodos , Humanos , Límite de Detección , Fragmentos de Péptidos/sangreAsunto(s)
Infecciones por Coronavirus/patología , Peptidil-Dipeptidasa A/genética , Neumonía Viral/patología , Serina Endopeptidasas/genética , Acoplamiento Viral , Internalización del Virus , Envejecimiento , Enzima Convertidora de Angiotensina 2 , Angiotensinas/sangre , Apelina/sangre , Betacoronavirus , Bradiquinina/sangre , COVID-19 , Catepsina B/metabolismo , Catepsina L/metabolismo , Humanos , Miocardio/patología , Miocitos Cardíacos/metabolismo , Pandemias , SARS-CoV-2RESUMEN
An activated plasma contact system is an abnormality observed in many Alzheimer's disease (AD) patients. Since mild cognitive impairment (MCI) patients often develop AD, we analyzed the status of contact system activation in MCI patients. We found that kallikrein activity, high molecular weight kininogen cleavage, and bradykinin levels- measures of contact system activation- were significantly elevated in MCI patient plasma compared to plasma from age- and education-matched healthy individuals. Changes were more pronounced in MCI patients with impaired short-term recall memory, indicating the possible role of the contact system in early cognitive changes.
Asunto(s)
Disfunción Cognitiva/sangre , Disfunción Cognitiva/diagnóstico , Trastornos de la Memoria/sangre , Trastornos de la Memoria/diagnóstico , Memoria a Corto Plazo/fisiología , Anciano , Biomarcadores/sangre , Bradiquinina/sangre , Disfunción Cognitiva/psicología , Estudios de Cohortes , Femenino , Humanos , Calicreínas/sangre , Quininógeno de Alto Peso Molecular/sangre , Masculino , Trastornos de la Memoria/psicología , Persona de Mediana Edad , Pruebas NeuropsicológicasAsunto(s)
Angioedemas Hereditarios , Bradiquinina/sangre , Calicreínas , Inhibidores de Serina Proteinasa/administración & dosificación , Administración Oral , Adulto , Angioedemas Hereditarios/sangre , Angioedemas Hereditarios/tratamiento farmacológico , Método Doble Ciego , Humanos , Calicreínas/antagonistas & inhibidores , Calicreínas/sangre , Masculino , Inhibidores de Serina Proteinasa/efectos adversosRESUMEN
Alzheimer's disease (AD) is characterized by the presence of proteinaceous brain deposits, brain atrophy, vascular dysfunction, and chronic inflammation. Along with cerebral inflammation, peripheral inflammation is also evident in many AD patients. Bradykinin, a proinflammatory plasma peptide, is also linked to AD pathology. For example, bradykinin infusion into the hippocampus causes learning and memory deficits in rats, and blockade of the bradykinin receptor lessens cognitive impairment in AD mouse models. Even though it has been hypothesized that plasma bradykinin could contribute to inflammation in AD, the level of plasma bradykinin and its association with beta-amyloid (Aß) pathology in AD patients had not been explored. Here, we assessed plasma bradykinin levels in AD patients and age-matched non-demented (ND) control individuals. We found significantly elevated plasma bradykinin levels in AD patients compared to ND subjects. Additionally, changes in plasma bradykinin levels were more profound in many AD patients with severe cognitive impairment, suggesting that peripheral bradykinin could play a role in dementia most likely via inflammation. Bradykinin levels in the cerebrospinal fluid (CSF) were reduced in AD patients and exhibited an inverse correlation with the CSF Aß40/Aß42 ratio. We also report that bradykinin interacts with the fibrillar form of Aß and co-localizes with Aß plaques in the post-mortem human AD brain. These findings connect the peripheral inflammatory pathway to cerebral abnormalities and identify a novel mechanism of inflammatory pathology in AD.
Asunto(s)
Enfermedad de Alzheimer/sangre , Bradiquinina/sangre , Disfunción Cognitiva/sangre , Anciano , Anciano de 80 o más Años , Enfermedad de Alzheimer/líquido cefalorraquídeo , Péptidos beta-Amiloides/sangre , Apolipoproteínas E/líquido cefalorraquídeo , Biomarcadores/sangre , Bradiquinina/líquido cefalorraquídeo , Estudios de Casos y Controles , Disfunción Cognitiva/líquido cefalorraquídeo , Progresión de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Placa Amiloide/sangreRESUMEN
BACKGROUND: Chronic spontaneous urticaria (CSU) is characterized by recurrent itchy weals and/or angioedema and is believed to be driven by mast cell activation. It was shown that excessive mast cell activation during anaphylaxis initiates contact activation, resulting in bradykinin release. Evidence for bradykinin release was never demonstrated in CSU. OBJECTIVE: To study biomarkers of bradykinin release in CSU. METHODS: Plasma samples of CSU patients were collected during routine visits at the outpatient clinic. Cleaved high molecular weight kininogen (cHK) was used as a biomarker for bradykinin release. cHK, factor XIIa-C1-inhibitor (FXIIa-C1-INH), kallikrein-C1-INH, plasmin-antiplasmin (PAP) complexes and soluble urokinase-type plasminogen activator receptor (suPAR) levels were determined by ELISA. Clinical data and data on tryptase levels were collected from medical records. cHK levels were compared to previously determined levels in hereditary angioedema (HAE). RESULTS: One hundred seventeen samples from 88 CSU patients and 28 samples from healthy controls were analysed. Median cHK level in CSU was 9.1% (range: 1.4%-21.5%), significantly increased compared to healthy controls (median 6.0% range: 0%-19.9%; P = .0005) and comparable to HAE (n = 46, median 10.3%, range 0%-44.3%, P > .9999). cHK levels normalized in patients during disease remission (median 6.5% range 1.5%-20.8%) but were not dependent on the presence of angioedema, acute angioedema attacks or response to antihistamines. Surprisingly, cHK levels were inversely correlated to serum tryptase (r = -0.65 P = .0137). C1-INH complexes and suPAR levels were not elevated in patients compared to healthy controls. PAP-complex levels in patients were elevated compared to healthy controls but there was no correlation between PAP-complex and cHK levels. CONCLUSIONS: cHK levels are elevated in symptomatic CSU patients compared to healthy controls, indicating increased bradykinin production. Increased cHK levels are not limited to patients with angioedema. CLINICAL RELEVANCE: If elevated bradykinin generation has clinical implications in the pathology of CSU is open to debate.
Asunto(s)
Bradiquinina , Urticaria Crónica , Adolescente , Adulto , Anciano , Biomarcadores/sangre , Bradiquinina/sangre , Bradiquinina/inmunología , Urticaria Crónica/sangre , Urticaria Crónica/inmunología , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
INTRODUCTION: Hereditary angioedema (HAE) is a rare autosomal dominant disease caused by genetic dysfunction of C1 inhibitor (C1-INH) due to mutations in the SERPING1 gene. The disorder is mediated mainly by bradykinin. The clinical course of the disease is varied and not related to genetic changes. OBJECTIVES: We aimed to evaluate redox homeostasis of peripheral blood mononuclear cells (PBMCs) in patients with HAE due to C1-INH deficiency (C1 INH HAE) by measuring the levels of reactive oxygen species (ROS) of PBMCs as well as plasma advanced glycation end products (AGEs) and advanced oxidation protein products (AOPPs). We also aimed to assess the effect of bradykinin on ROS levels. PATIENTS AND METHODS: We enrolled 30 adults with C1-INH-HAE and 15 healthy individuals. The levels of ROS were measured by flow cytometry, while the plasma levels of AGEs and AOPPs were determined spectrophotometrically by enzyme linked immunosorbent assays. RESULTS: Basal and hydrogen peroxide (H2O2)-induced ROS levels were higher in patients with HAE when compared with controls (P = 0.002 and P = 0.001, respectively), indicating abnormalities in redox homeostasis. Plasma AOPP and AGE levels were similar in both groups. Bradykinin reduced basal and H2O2-induced ROS generation in PBMCs only in patients with HAE (P = 0.03). CONCLUSIONS: The higher basal and H2O2-induced ROS levels in patients with C1 INH HAE indicate redox imbalance. However, by reducing basal and H2O2-induced ROS levels, bradykinin shows antioxidant action in this disorder.
Asunto(s)
Angioedemas Hereditarios/metabolismo , Bradiquinina/sangre , Proteína Inhibidora del Complemento C1/genética , Leucocitos Mononucleares/metabolismo , Estrés Oxidativo , Adulto , Angioedemas Hereditarios/sangre , Angioedemas Hereditarios/genética , Bradiquinina/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , MutaciónRESUMEN
Hereditary autoinflammatory diseases are caused by gene mutations of the innate immune pathway, e.g. nucleotide receptor protein 3 (NLRP3). Here, we report a four-generation family with cold-induced urticarial rash, arthralgia, chills, headache and malaise associated with an autosomal-dominant inheritance. Genetic studies identify a substitution mutation in gene F12 (T859A, resulting in p.W268R) which encodes coagulation factor XII (FXII). Functional analysis reveals enhanced autocatalytic cleavage of the mutated protein and spontaneous FXII activation in patient plasma and in supernatant of transfected HEK293 cells expressing recombinant W268R-mutated proteins. Furthermore, we observe reduced plasma prekallikrein, cleaved high molecular weight kininogen and elevated plasma bradykinin. Neutrophils are identified as a local source of FXII. Interleukin-1ß (IL-1ß) is upregulated in lesional skin and mononuclear donor cells exposed to recombinant mutant proteins. Treatment with icatibant (bradykinin-B2-antagonist) or anakinra (interleukin-1-antagonist) reduces disease activity in patients. In conclusion, our findings provide a link between contact system activation and cytokine-mediated inflammation.
Asunto(s)
Frío/efectos adversos , Factor XII/metabolismo , Enfermedades Autoinflamatorias Hereditarias/metabolismo , Adulto , Coagulación Sanguínea , Bradiquinina/análogos & derivados , Bradiquinina/sangre , Bradiquinina/uso terapéutico , Factor XII/genética , Femenino , Células HEK293 , Enfermedades Autoinflamatorias Hereditarias/genética , Enfermedades Autoinflamatorias Hereditarias/patología , Humanos , Mediadores de Inflamación , Proteína Antagonista del Receptor de Interleucina 1/uso terapéutico , Interleucina-1beta/metabolismo , Quininógeno de Alto Peso Molecular/metabolismo , Masculino , Persona de Mediana Edad , Mutación , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Neutrófilos , Linaje , Fenotipo , Calicreína Plasmática/metabolismo , Proteínas Recombinantes , Piel/patologíaRESUMEN
OBJECTIVE: To explore the effect of bradykinin on rats with thromboangiitis obliterans (TAO) through the phosphatidylinositol 3-hydroxy kinase/protein kinase B (PI3K/Akt) signaling pathway. MATERIALS AND METHODS: The female Wistar rats were injected with lauric acid via the femoral artery to establish the TAO model, and they were randomly divided into control group (healthy rats), model group (TAO rats) and bradykinin group (TAO rats injected with bradykinin B2 receptor-specific inhibitor). The control was set in each group before the operation. The level of serum bradykinin in each group was detected via enzyme-linked immunosorbent assay (ELISA), and the reactive oxygen species (ROS) level, Caspase-3 activity and PI3K/Akt protein concentration in vascular tissues were measured via ELISA, Western blotting, ROS assay, and Caspase-3 activity assay, respectively. Moreover, the specific therapeutic mechanism of bradykinin was analyzed. RESULTS: In control group, the intima of the lower extremity venous tissues was smooth, the extima had no evident changes, and there was no inflammatory cell invasion around the arteries and veins. In model group, there was massive inflammatory cell invasion into the lower extremity venous tissues. In bradykinin group, fibrosis and atrophy occurred in venous tissues, the extima was thickened without fibrosis, and there was phagocytosis of neutrophils and mononuclear macrophages around the arteries and veins, as well as massive inflammatory infiltration. The PI3K/Akt protein concentration in lower extremity venous tissues was the highest in control group and the lowest in bradykinin group, and there were statistically significant differences (p<0.01). At 24 h after administration of doxorubicin (DOX), the level of ROS in lower extremity venous tissues was higher in bradykinin group than that in model group (p<0.05), and it was also higher in model group than that in control group (p<0.05). Besides, the activity of Caspase-3 in lower extremity venous tissues was significantly increased in bradykinin group compared with that in model group and control group, while it was slightly higher in model group than that in control group (p<0.05). CONCLUSIONS: The low expression of bradykinin can promote TAO in rats by the mechanism that it inhibits the PI3K/Akt signaling pathway to raise the oxidative stress level, thereby aggravating TAO.
Asunto(s)
Antagonistas del Receptor de Bradiquinina B2/administración & dosificación , Bradiquinina/sangre , Transducción de Señal/efectos de los fármacos , Tromboangitis Obliterante/tratamiento farmacológico , Vasodilatadores/administración & dosificación , Animales , Apoptosis/efectos de los fármacos , Antagonistas del Receptor de Bradiquinina B2/farmacología , Femenino , Ácidos Láuricos/efectos adversos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Tromboangitis Obliterante/inducido químicamente , Tromboangitis Obliterante/metabolismo , Vasodilatadores/farmacologíaRESUMEN
Bradykinin is a proinflammatory factor that mediates angioedema and inflammation in many diseases. It is a key player in some types of hereditary angioedema and is involved in septic shock, traumatic injury, Alzheimer's disease (AD), and stroke, among others. Activation of the plasma contact system leads to elevated levels of plasma kallikrein, which cleaves high molecular weight kininogen (HK) to release bradykinin. Drug development for bradykinin-meditated pathologies has focused on designing inhibitors to the enzymes that cleave HK (to prevent bradykinin release) or antagonists of endothelial bradykinin receptors (to prevent downstream bradykinin action). Here we show a strategy to block bradykinin generation by using an HK antibody that binds to HK, preventing its cleavage and subsequent bradykinin release. We show that this antibody blocks dextran sodium sulfate-induced HK cleavage and bradykinin production. Moreover, while the pathogenic AD peptide ß-amyloid (Aß)42 cleaves HK and induces a dramatic increase in bradykinin production, our HK antibody blocked these events from occurring. These results may provide strategies for developing treatments for bradykinin-driven pathologies.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Péptidos beta-Amiloides/metabolismo , Anticuerpos/administración & dosificación , Bradiquinina/metabolismo , Quininógeno de Alto Peso Molecular/antagonistas & inhibidores , Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/metabolismo , Bradiquinina/sangre , Humanos , Quininógeno de Alto Peso Molecular/metabolismoRESUMEN
Hereditary angioedema has been attributed to an inherited deficiency of C1 esterase inhibitor that increases vascular permeability. The role of C1 esterase inhibitor in burn patients has not been described previously. In this study, we attempted to identify the relationship between serial changes of C1 esterase inhibitor activity and the clinical course in major burn patients. This study was a single-center, prospective, observational study. C1 esterase inhibitor activity values were serially examined in major burn patients admitted into the burn center from April 2014 to December 2016. Inclusion criteria were age ≥16 years old and %TBSA burned ≥20%. This study included 38 patients with major burn. C1 esterase inhibitor activity after burn dropped acutely on days 1 and 2 but increased immediately until days 3 to 5, after which it continued to gradually increase to above the reference value. C1 esterase inhibitor activity on admission showed significant inverse correlation with the volume of infusion per body weight required in the first 24 hours after injury and %TBSA burned (r = -0.405, P = 0.01; r = -0.375, P = 0.02, respectively). C1 esterase inhibitor activity on admission was significantly lower in the nonsurvivors than in the survivors during the 28-day evaluation period (59% vs 90%, P = 0.01). These findings suggest that C1 esterase inhibitor may play a critical role in regulating vascular permeability in the acute phase following the burn injury.
Asunto(s)
Quemaduras/sangre , Proteína Inhibidora del Complemento C1/análisis , APACHE , Adulto , Anciano , Biomarcadores/sangre , Bradiquinina/sangre , Permeabilidad Capilar , Estudios de Casos y Controles , Femenino , Fluidoterapia , Hemodinámica , Humanos , Masculino , Persona de Mediana Edad , Puntuaciones en la Disfunción de Órganos , Estudios Prospectivos , Resucitación , Lactato de Ringer/administración & dosificación , Albúmina Sérica/análisisRESUMEN
OBJECTIVE: We recently investigated the pathways of immunoreactive bradykinin (iBK) formation in fresh blood of normal volunteers and of patients with hereditary angioedema due to C1-esterase inhibitor deficiency (HAE-1/-2). Herein, we adapted the techniques to small volumes (200 µl) of previously frozen citrated plasma and further analyzed the mechanisms of iBK formation with additional biotechnological inhibitors. RESULTS: Measurable iBK formation was observed under stimulation with tissue kallikrein (KLK-1, 10 nM), the particulate material Kontact-APTT (concentration reduced to 2% v/v) or recombinant tissue plasminogen activator (tPA, 169 nM), with little background in unstimulated plasma incubated for up to 2 h. Plasma samples from HAE-1/-2 patients responded earlier to tPA than those from controls, as previously reported with whole blood. Lanadelumab inhibited iBK formation induced by Kontact-APTT and tPA. A highly specific plasmin inhibitor, DX-1000, abolished tPA-induced iBK formation in plasma but had no effect against Kontact-APTT, confirming the role of fibrinolysis in tPA-induced kinin formation. The anti-lanadelumab neutralizing antibody M293-D02 reversed the inhibitory effects of lanadelumab. Frozen plasma is a suitable material for measuring iBK formation kinetics, with possible applications such as investigating the effect of rare disease states on the kallikrein-kinin system and monitoring the effect of HAE prophylactic treatments.
Asunto(s)
Bradiquinina/biosíntesis , Fibrinólisis/fisiología , Angioedema Hereditario Tipos I y II/sangre , Calicreínas/química , Activador de Tejido Plasminógeno/química , Adulto , Anticuerpos Monoclonales Humanizados/química , Anticuerpos Neutralizantes/química , Antifibrinolíticos/química , Recolección de Muestras de Sangre/métodos , Bradiquinina/sangre , Estudios de Casos y Controles , Femenino , Fibrinolisina/antagonistas & inhibidores , Fibrinolisina/metabolismo , Congelación , Humanos , Masculino , Plasma/química , Proteínas Recombinantes/químicaRESUMEN
Kallikrein is the key contact system mediator responsible for the conversion of high-molecular-weight kininogen into the inflammatory vasodilator peptide bradykinin, a process regulated by C1-esterase inhibitor (C1-INH). In hereditary angioedema (HAE), genetic mutations result in deficient or dysfunctional C1-INH and dysregulation of the contact system leading to recurrent, sometimes fatal, angioedema attacks. IONIS-PKKRx is a second-generation 2'-O-(2-methoxyethyl)-modified chimeric antisense oligonucleotide, designed to bind and selectively reduce prekallikrein (PKK) mRNA in the liver. IONIS-PKKRx demonstrated dose-dependent reduction of human prekallikrein hepatic mRNA and plasma protein in transgenic mice and dose- and time-dependent reductions of plasma PKK in Cynomolgus monkeys. Similar dose-dependent reductions of plasma PKK levels were observed in healthy human volunteers accompanied by decreases in bradykinin generation capacity with an acceptable safety and tolerability profile. These results highlight a novel and specific approach to target PKK for the treatment of HAE and other diseases involving contact system activation and overproduction of bradykinin.
Asunto(s)
Angioedemas Hereditarios/terapia , Bradiquinina/genética , Complemento C1s/genética , Precalicreína/genética , Angioedemas Hereditarios/sangre , Angioedemas Hereditarios/genética , Animales , Animales Modificados Genéticamente/sangre , Bradiquinina/sangre , Proteína Inhibidora del Complemento C1/farmacología , Complemento C1s/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Voluntarios Sanos , Humanos , Hígado/efectos de los fármacos , Hígado/metabolismo , Macaca fascicularis/sangre , Ratones , Oligodesoxirribonucleótidos Antisentido/genética , Oligodesoxirribonucleótidos Antisentido/farmacología , Precalicreína/antagonistas & inhibidoresRESUMEN
BACKGROUND: Bradykinin is an important mediator of inflammation and vascular permeability and could have an important role in the development of septic shock. Measurement of bradykinin by immunological methods may suffer from interference and lack of specificity. We developed and validated a liquid chromatography mass spectrometry assay (LC-MS/MS) for plasma bradykinin. METHODS: We used plasma samples from healthy volunteers (nâ¯=â¯19) and patients with septic shock (nâ¯=â¯47). Stable isotope bradykinin internal standard was added to samples before solid-phase extraction and quantification by LC-MS/MS. Stability of bradykinin was studied for 12â¯months. RESULTS: Our assay has good sensitivity (0.1â¯nmol/l) and a wide linear range (0.1-1000â¯nmol/l). Bradykinin added to plasma was stable for 12â¯months at -20⯰C when a mixture of protease inhibitors was added at sampling but degraded during repeated freezing and thawing. Bradykinin concentration in plasma from septic shock patients (<0.1-0.6â¯nmol/l) did not change significantly during shock and recovery but differed slightly from that in healthy individuals (0.5-1.1â¯nmol/l). CONCLUSIONS: Our bradykinin assay was successfully used to determine bradykinin concentrations in plasma samples. Intensive care unit patients with septic shock had low concentrations of plasma bradykinin during both shock and recovery phases.
Asunto(s)
Bradiquinina/sangre , Choque Séptico/sangre , Adulto , Anciano , Anciano de 80 o más Años , Cromatografía Liquida , Femenino , Humanos , Masculino , Persona de Mediana Edad , Espectrometría de Masas en TándemRESUMEN
Angioedema is a rare adverse effect of the commonly used angiotensin-converting enzyme inhibitors (ACEi) and is reported to occur with a prevalence of 0.1%-0.7%. Although most ACEi-induced angioedema (ACEi-A) cases are mild, severe cases requiring intensive care and even resulting in death have been reported in the literature. The mechanisms underlying ACEi-A are not yet fully understood, but bradykinin and/or substance P accumulation resulting from inhibition of ACE is believed to play a crucial role. ACEi-A occurs at variable frequencies across different racial groups, suggesting a genetic association with the development of ACEi-A. To date, one genome-wide association study and several candidate gene studies have been published on the association of genetic variation with ACEi-A. Genetic associations reported have been attributed to several distinct mechanisms: (a) genes coding for alternative enzymes responsible for the degradation of bradykinin and/or substance P in the diminution of ACE activity (b) ACE gene function, (c) bradykinin receptor genes, (d) genes implicated in immune and inflammation regulation and (e) genes in the fibrinolytic and coagulation pathway. Despite several plausible genetic associations, there are currently no genetic variants with sufficient effect to be clinically useful. The low incidence of ACEi-A suggests that a combination of genomic approaches with the capability to detect potentially important variants might be required to shed light on the mechanism of this adverse reaction. Additionally, many non-genetic risk factors associated with ACEi-A suggest the potential contribution of epigenetic dysregulation.
Asunto(s)
Angioedema , Inhibidores de la Enzima Convertidora de Angiotensina/efectos adversos , Polimorfismo Genético , Angioedema/sangre , Angioedema/inducido químicamente , Angioedema/epidemiología , Angioedema/genética , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Bradiquinina/sangre , Bradiquinina/genética , Estudio de Asociación del Genoma Completo , Humanos , Farmacogenética , Prevalencia , Sustancia P/sangre , Sustancia P/genéticaRESUMEN
OBJECTIVE: To investigate the expression and time-dependent manner of bradykinin (BK) as well as its receptors (Bradykinin receptors 1 and 2, B1R and B2R) in spinal cord ischemia-reperfusion injury (SCII) in rat model. METHODS: Sprague-Dawley (SD) rats were subjected to 1â¯h of infra-renal abdominal aorta occlusion and reperfused for 3â¯h to 5â¯d to induce SCII. The concentration of BK in serum was detected by enzyme linked immunosorbent assay (ELISA). In situ expression of BK receptors was evaluated by immunochemistry and their mRNA level was evaluated by Real time quantitative-PCR (RTq-PCR). RESULTS: The concentration of BK in serum was increased right after following SCII. Both of the BK receptors were detected and up-regulated in 24â¯h and 48â¯h after injury. The levels of B1R and B2R mRNA were up-regulated after SCII, and the B1R mRNA dropped to basal level after 6â¯h, but B2R mRNA dropped to lower level right after injury, peaked at 3â¯h, then remained a lower level from 6â¯h till 5â¯day. CONCLUSION: This study provides the evidence of the expression of BK and its receptors in SCII in rat model, and suggests that BK and its receptors may have some physiological or pathological significance in SCII.
Asunto(s)
Biomarcadores/metabolismo , Bradiquinina/sangre , Modelos Animales de Enfermedad , Receptor de Bradiquinina B1/metabolismo , Receptor de Bradiquinina B2/metabolismo , Daño por Reperfusión/diagnóstico , Isquemia de la Médula Espinal/diagnóstico , Animales , Masculino , Ratas , Ratas Sprague-Dawley , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B2/genética , Daño por Reperfusión/metabolismo , Isquemia de la Médula Espinal/metabolismoRESUMEN
The plasma proteins factor XII (FXII) and prekallikrein (PK) undergo reciprocal activation to the proteases FXIIa and kallikrein by a process that is enhanced by surfaces (contact activation) and regulated by the serpin C1 inhibitor. Kallikrein cleaves high-molecular-weight kininogen (HK), releasing the vasoactive peptide bradykinin. Patients with hereditary angioedema (HAE) experience episodes of soft tissue swelling as a consequence of unregulated kallikrein activity or increased prekallikrein activation. Although most HAE cases are caused by reduced plasma C1-inhibitor activity, HAE has been linked to lysine/arginine substitutions for Thr309 in FXII (FXII-Lys/Arg309). Here, we show that FXII-Lys/Arg309 is susceptible to cleavage after residue 309 by coagulation proteases (thrombin and FXIa), resulting in generation of a truncated form of FXII (δFXII). The catalytic efficiency of δFXII activation by kallikrein is 15-fold greater than for full-length FXII. The enhanced rate of reciprocal activation of PK and δFXII in human plasma and in mice appears to overwhelm the normal inhibitory function of C1 inhibitor, leading to increased HK cleavage. In mice given human FXII-Lys/Arg309, induction of thrombin generation by infusion of tissue factor results in enhanced HK cleavage as a consequence of δFXII formation. The effects of δFXII in vitro and in vivo are reproduced when wild-type FXII is bound by an antibody to the FXII heavy chain (HC; 15H8). The results contribute to our understanding of the predisposition of patients carrying FXII-Lys/Arg309 to angioedema after trauma, and reveal a regulatory function for the FXII HC that normally limits PK activation in plasma.