Bradykinin Receptor Research Articles

The role of vasoactive peptides in skin homeostasis-focus on adiponectin and the kallikrein-kinin system.

Vasoactive peptides often serve a multitude of functions aside from their direct effects on vasodynamics. This article will review the existing literature on two vasoactive peptides and their involvement in skin homeostasis: adiponectin and-as the main representative of the kallikrein-kinin system-bradykinin. Adiponectin is the most abundantly expressed adipokine in the human organism, where it is mainly localized in fat depots including subcutaneous adipose tissue, from where adiponectin can exert paracrine effects. The involvement of adiponectin in skin homeostasis is supported by a number of studies reporting the effects of adiponectin in isolated human keratinocytes, sebocytes, fibroblasts, melanocytes, and immune cells. Regarding skin pathology, the potential involvement of adiponectin in psoriasis, atopic dermatitis, scleroderma, keloid, and melanogenesis is discussed in this article. The kallikrein-kinin system is composed of a variety of enzymes and peptides, most of which have been identified to be expressed in the skin. This also includes the expression of bradykinin receptors on most skin cells. Bradykinin is one of the very few hormones that is targeted by treatment in routine clinical use in dermatology-in this case for the treatment of hereditary angioedema. The potential involvement of bradykinin in wound healing, psoriasis, and melanoma is further discussed in this article. This review concludes with a call for additional preclinical and clinical studies to further explore the therapeutic potential of adiponectin supplementation (for psoriasis, atopic dermatitis, wound healing, scleroderma, and keloid) or pharmacological interference with the kallikrein-kinin system (for wound healing, psoriasis, and melanoma).

Angiotensin receptor-neprilysin inhibitor improves coronary collateral perfusion.

We investigated the pleiotropic effects of an angiotensin receptor-neprilysin inhibitor (ARNi) on collateral-dependent myocardial perfusion in a rat model of coronary arteriogenesis, and performed comprehensive analyses to uncover the underlying molecular mechanisms. A rat model of coronary arteriogenesis was established by implanting an inflatable occluder on the left anterior descending coronary artery followed by a 7-day repetitive occlusion procedure (ROP). Coronary collateral perfusion was measured by using a myocardial particle infusion technique. The putative ARNi-induced pro-arteriogenic effects were further investigated and compared with an angiotensin-converting enzyme inhibitor (ACEi). Expression of the membrane receptors and key enzymes in the natriuretic peptide system (NPS), renin-angiotensin-aldosterone system (RAAS) and kallikrein-kinin system (KKS) were analyzed by quantitative polymerase chain reaction (qPCR) and immunoblot assay, respectively. Protein levels of pro-arteriogenic cytokines were measured by enzyme-linked immunosorbent assay, and mitochondrial DNA copy number was assessed by qPCR due to their roles in arteriogenesis. Furthermore, murine heart endothelial cells (MHEC5-T) were treated with a neprilysin inhibitor (NEPi) alone, or in combination with bradykinin receptor antagonists. MHEC5-T proliferation was analyzed by colorimetric assay. The in vivo study showed that ARNis markedly improved coronary collateral perfusion, regulated the gene expression of KKS, and increased the concentrations of relevant pro-arteriogenic cytokines. The in vitro study demonstrated that NEPis significantly promoted MHEC5-T proliferation, which was diminished by bradykinin receptor antagonists. ARNis improve coronary collateral perfusion and exert pro-arteriogenic effects via the bradykinin receptor signaling pathway.

Development of a multigram synthesis of the bradykinin receptor 2 agonist FR-190997 and analogs thereof.

Using Fujisawa's B2R agonist FR-190997, we recently demonstrated for the first time that agonism at the bradykinin receptor type 2 (B2R) produces substantial antiproliferative effects. FR-190997 elicited an EC50 of 80 nM in the triple-negative breast cancer cell line MDA-MB-231, a much superior performance to that exhibited by most approved breast cancer drugs. Consequently, we initiated a program aiming primarily at synthesizing adequate quantities of FR-190997 to support further in vitro and in vivo studies toward its repurposing for various cancers and, in parallel, enable the generation of novel FR-190997 analogs for an SAR study. Prerequisite for this endeavor was to address the synthetic challenges associated with the FR-190997 scaffold, which the Fujisawa chemists had constructed in 20 steps, 13 of which required chromatographic purification. We succeeded in developing a 17-step synthesis amenable to late-stage diversification that eliminated all chromatography and enabled access to multigram quantities of FR-190997 and novel derivatives thereof, supporting further anticancer research based on B2R agonists.

Bradykinin B1 receptor antagonist protects against cold stress-induced erectile dysfunction in rats.

Erectile dysfunction (ED) demonstrates seasonal variation with higher rates in winter, and we hypothesize that endothelial damage in erectile tissue caused by bradykinin receptor B1 (B1R) might be detrimental to this change. To find out direct correlations between cold stress and ED, through which to further investigate the functional roles of B1R in erectile tissue and to elucidate the therapeutic roles of the B1R antagonist in a cold stress-induced ED rat model. Cold stress rat models are established through long-term intermittent exposure to low temperature. After their erectile function was assessed, ED rats were treated with the B1R antagonist through intraperitoneal injection. Penile tissues were obtained at the end of the experiment after measurement of intracavernosal pressure/mean arterial pressure (ICP/MAP); the location and distribution of cytokine expression were determined by immunohistochemistry; cytokine levels and NOS and CD31 expression were detected by Western blotting; and collagen fibers and smooth muscles were observed through Masson staining. Cold stress impairs erectile function, and the B1R antagonist protects against it. We observed decreased erection frequency, prolonged erection latency time, decreased ICP/MAP, overexpression of B1R, increased expression of cytokines on cavernous sinus endothelium, and increased levels of collagen fibers/smooth muscles on erectile tissue in response to cold stress. Also, NOS and CD31 expression was downregulated. B1R antagonist treatment shows enhanced erectile function through increased erection frequency, shortened erection latency time, and increased ICP/MAP. Also, it reduces collagen fibers/smooth muscles, TNF-α, TGF-β1, and IL-6 and upregulates the expression of nNOS and CD31. Our findings cast new light on the correlations between cold stress and erectile function and potential new applications of existing B1R antagonist drugs in the field of ED. Our data support that cold stress impairs erectile function. B1R-mediated, cytokine-induced corpus cavernosum fibrosis and endothelial damage might be the main reason behind it, and B1R inhibition protects against fibrosis and endothelial damage. Other ways of B1R antagonist blocking methods in different types of ED still need to be investigated. Long-term intermittent cold stress impairs erectile function, and B1R-mediated, cytokine-induced corpus cavernosum fibrosis and endothelial damage might be the main reason behind it. B1R inhibition also protects against fibrosis and endothelial damage. Our data support the hypothesis that cold stress impairs erectile function and that B1R blockade ameliorates the symptoms of ED, possibly by reversing fibrosis and endothelial damage in erectile tissue.

Inhibition of angiotensin converting enzyme increases PKCβI isoform expression via activation of substance P and bradykinin receptors in cultured astrocytes of mice.

Angiotensin-converting enzyme inhibitor (ACEi) inhibits the catalysis of angiotensin I to angiotensin II and the degradation of substance P (SP) and bradykinin (BK). While the possible relationship between ACEi and SP in nociceptive mice was recently suggested, the effect of ACEi on signal transduction in astrocytes remains unclear. This study examined whether ACE inhibition with captopril or enalapril modulates the levels of SP and BK in primary cultured astrocytes and whether this change modulates PKC isoforms (PKCα, PKCβI, and PKCε) expression in cultured astrocytes. Immunocytochemistry and Western blot analysis were performed to examine the changes in the levels of SP and BK and the expression of the PKC isoforms in primary cultured astrocytes, respectively. The treatment of captopril or enalapril increased the immunoreactivity of SP and BK significantly in glial fibrillary acidic protein-positive cultured astrocytes. These increases were suppressed by a pretreatment with an angiotensin-converting enzyme. In addition, treatment with captopril increased the expression of the PKCβI isoform in cultured astrocytes, while there were no changes in the expression of the PKCα and PKCε isoforms after the captopril treatment. The captopril-induced increased expression of the PKCβI isoform was inhibited by a pretreatment with the neurokinin-1 receptor antagonist, L-733,060, the BK B1 receptor antagonist, R 715, or the BK B2 receptor antagonist, HOE 140. These results suggest that ACE inhibition with captopril or enalapril increases the levels of SP and BK in cultured astrocytes and that the activation of SP and BK receptors mediates the captopril-induced increase in the expression of the PKCβI isoform.

Фармакокинетика антагониста брадикинина производного 1,4-бензодиазепин-2-она в эксперименте

Introduction. The 1,4-benzodiazepin-2-one derivative (codenamed – PAV-0056), in a wide range of low doses 0.1–10 mg/kg has a pronounced analgesic effect as bradykinin receptor type 1 antagonist. Preclinical study of the pharmacokinetics of a new potential analgesic with different routes of administration in appropriate doses seems to be relevant. Aim. To evaluate the pharmacokinetic parameters, bioavailability and transmembrane transport of the 1,4-benzodiazepin-2-one derivative PAV-0056. Materials and methods. PAV-0056 at doses of 0.1, 1 and 10 mg/kg was administered in a 1% aqueous solution of polyvinylpyrrolidone into the stomach and vein of male Sprague Dawley rats (age 1.5–3 months) weighing 250–300 g. We determined the plasma concentration of PAV-0056 in the blood of rats at discrete time intervals using mass spectrometry. The pharmacokinetic parameters and absolute bioavailability of PAV-0056 were studied. The permeability of PAV-0056 through a cell monolayer was studied on a 21-day cell culture originated from of colorectal carcinoma (CaCO-2). Results. The absolute bioavailability of the 1,4-benzodiazepin-2-one derivative PAV-0056 after administration into the stomach of rats at a dose of 0.1 mg/kg is 10.1 ± 6.1%, at a dose of 1 mg/kg – 2.2 ± 0.1%, at a dose of 10 mg/kg – 6.4 ± 0.8%. Systemic exposure AUC and maximum plasma concentration Cmax change linearly with increasing dose of PAV-0056 from 0.1 to 10 mg/kg (coefficients of determination R2 are 0.9767 and 0.9976, respectively). PAV-0056 is absorbed from the small intestine within 1 h and eliminated from plasma within 5 h (the lower limit of quantification of PAV-0056 in plasma is 0.1 ng/ml). After intravenous administration, PAV-0056 is eliminated within 0.5 h. The analgesic has a total clearance (ClT) of 83.16 l/kg/h and is not a P-glycoprotein substrate. Conclusion. The lipophilic derivative of 1,4-benzodiazepin-2-one, PAV-0056 after administration into the stomach is absorbed from the small intestine by passive diffusion with a bioavailability of no more than 10.1 ± 6.1%. Linear pharmacokinetics and no interaction with P-glycoprotein allow PAV-0056 to be positioned as a non-opioid analgesic with a good safety profile.

Slučaj angioedema hereditarnog porekla u seoskoj ambulanti nadomak Subotice

Introduction: Hereditary angioedema (HAE) is inherited in an autosomal dominant manner. It manifests with bradykinin-mediated swelling due to a deficiency of C1 inhibitor. Managing these patients is challenging during acute edema and the onset of asphyxiation. Transportation to the hospital takes 25 minutes. Objective: To present a patient with HAE for whom specific therapy has only recently become available. Case Presentation: A female patient presented with difficulty breathing, swelling of the lips, and periorbital edema. Status: visibly distressed, talkative, facial and lip swelling, visible mucosa without edema, uvula and pharynx calm. Respiratory movements symmetric, without the use of accessory muscles. Auscultation revealed normal breath sounds, tachycardic heart action, clear tones without murmurs, and no urticaria. Blood pressure 120/70 mmHg, heart rate 100/min., respiratory rate 30/min., SpO2 98%, capillary refill time (CRT) 2 sec. Treatment: infusion, Methylprednisolone 40 mg IV. She was transported with monitoring. During transport, the condition worsened: blood pressure 105/60 mmHg, heart rate 130/min., respiratory rate 40/min., SpO2 94%, CRT 2 sec. Oxygen at 15 L/min. was initiated, an additional IV line was opened, and capnography showed ETCO2 of 36 mmHg. Methylprednisolone was increased to 80 mg. Preparation for difficult intubation was initiated. Parameters did not worsen, and the patient was transferred to the emergency department of the General Hospital Subotica. Conclusion: Since specific therapies, such as human C1 inhibitor concentrate or bradykinin receptor antagonists, have been approved for self-administration at home during acute attacks, the treatment of these patients has been facilitated, minimizing the potential fatal outcomes and morbidity.

Altered Left Ventricular Rat Gene Expression Induced by the Myosin Activator Omecamtiv Mecarbil.

To explore the impact of omecamtiv mecarbil (OM) on the gene expression profile in adult male rats. Fourteen male Wistar rats were randomly assigned to a single OM (1.2 mg/kg/h; n = 6) or placebo (n = 8) 30-min infusion. Echocardiography was performed before and after OM infusion. Seven days after infusion, rats were euthanized, and left ventricular (LV) tissues were removed for real-time quantitative polymerase chain reaction (RTq-PCR) experiments. After OM infusion, pro-apoptotic Bax-to-Bcl2 ratio was decreased, with increased Bcl2 and similar Bax gene expression. The gene expression of molecules regulating oxidative stress, including glutathione disulfide reductase (Gsr) and superoxide dismutases (Sod1/Sod2), remained unchanged, whereas the expression of antioxidant glutathione peroxidase (Gpx) increased. While LV gene expression of key energy sensors, peroxisome proliferator activator (Ppar) α and γ, AMP-activated protein kinase (Ampk), and carnitine palmitoyltransferase 1 (Cpt1) remained unchanged after OM infusion, and the expression of pyruvate dehydrogenase kinase 4 (Pdk4) increased. The LV expression of the major myocardial glucose transporter Glut1 decreased, with no changes in Glut4 expression, whereas the LV expression of oxidized low-density lipoprotein receptor 1 (Olr1) and arachidonate 15-lipoxygenase (Alox15) increased, with no changes in fatty acid transporter Cd36. An increased LV expression of angiotensin II receptors AT1 and AT2 was observed, with no changes in angiotensin I-converting enzyme expression. The Kalikrein-bradykinin system was upregulated with increased LV expression of kallikrein-related peptidases Klk8, Klk1c2, and Klk1c12 and bradykinin receptors B1 and B2 (Bdkrb1 and Bdkrb2), whereas the LV expression of inducible nitric oxide synthase 2 (Nos2) increased. LV expression in major molecular determinants involved in calcium-dependent myocardial contraction remained unchanged, except for an increased LV expression of calcium/calmodulin-dependent protein kinase II delta (Cacna1c) in response to OM. A single intravenous infusion of OM, in adult healthy rats, resulted in significant changes in the LV expression of genes regulating apoptosis, oxidative stress, metabolism, and cardiac contractility.

The des-Arg9-bradykinin/B1R axis: Hepatic damage in COVID-19.

Patients infected by the SARS-CoV-2 virus are commonly diagnosed with threatening liver conditions associated with drug-induced therapies and systemic viral action. RNA-Seq data from cells in bronchoalveolar lavage fluid from COVID-19 patients have pointed out dysregulation of kallikrein-kinin and renin-angiotensin systems as a possible mechanism that triggers multi-organ damage away from the leading site of virus infection. Therefore, we measured the plasma concentration of biologically active peptides from the kallikrein-kinin system, bradykinin and des-Arg9-bradykinin, and liver expression of its proinflammatory axis, bradykinin 1 receptor (B1R). We measured the plasma concentration of bradykinin and des-Arg9-bradykinin of 20 virologically confirmed COVID-19 patients using a liquid chromatography-tandem mass spectrometry-based methodology. The expression of B1R was evaluated by immunohistochemistry from post-mortem liver specimens of 27 COVID-19 individuals. We found a significantly higher blood level of des-Arg9-bradykinin and a lower bradykinin concentration in patients with COVID-19 compared to a healthy, uninfected control group. We also observed increased B1R expression levels in hepatic tissues of patients with COVID-19 under all hepatic injuries analyzed (liver congestion, portal vein dilation, steatosis, and ischemic necrosis). Our data indicate that des-Arg9-bradykinin/B1R is associated with the acute hepatic dysfunction induced by the SARS-CoV-2 virus infection in the pathogenesis of COVID-19.

Bradykinin 2 receptors contribute to the exaggerated exercise pressor reflex in a rat model of simulated peripheral artery disease.

We investigated the role played by bradykinin 2 (B2) receptors in the exaggerated exercise pressor reflex in rats with a femoral artery ligated for 72 h to induce simulated peripheral artery disease (PAD). We hypothesized that in decerebrate, unanesthetized rats with a ligated femoral artery, hindlimb arterial injection of HOE-140 (100 ng, B2 receptor antagonist) would reduce the pressor response to 30 s of electrically induced 1 Hz hindlimb skeletal muscle contraction, and 30 s of 1 Hz hindlimb skeletal muscle stretch (a model of mechanoreflex activation isolated from contraction-induced metabolite production). We hypothesized no effect of HOE-140 in sham-operated "freely perfused" rats. In both freely perfused (n = 4) and "ligated" (n = 4) rats, we first confirmed efficacious B2 receptor blockade by demonstrating that HOE-140 injection significantly reduced (P < 0.05) the peak increase in mean arterial pressure (peak ΔMAP) in response to hindlimb arterial injection of bradykinin. In subsequent experiments, we found that HOE-140 reduced the peak ΔMAP response to muscle contraction in ligated (n = 14; control: 23 ± 2; HOE-140: 17 ± 2 mmHg; P = 0.03) but not freely perfused rats (n = 7; control: 17 ± 3; HOE-140: 18 ± 4 mmHg; P = 0.65). Furthermore, HOE-140 had no effect on the peak ΔMAP response to stretch in ligated rats (n = 14; control: 37 ± 4; HOE-140: 32 ± 5 mmHg; P = 0.13) but reduced the integrated area under the blood pressure signal over the final ∼20 s of the maneuver. The data suggest that B2 receptors contribute to the exaggerated exercise pressor reflex in rats with simulated PAD, and that contribution includes a modest role in the chronic sensitization of the mechanically activated channels/afferents that underlie mechanoreflex activation.

Reported estimation techniques for quantification of zaltoprofen: A review

Zaltoprofen is a nonsteroidal anti-inflammatory drug used as an analgesic, antipyretic, and antinflammatory agent. It is a selective COX-2 inhibitor and also inhibits bradykinin-induced pain responses without blocking bradykinin receptors. Zaltoprofen is prescribed for treating pain and inflammatory conditions caused by various disorders such as dental pain, musculoskeletal pain, postoperative pain, and lumbar pain, and frozen shoulder, osteoarthritis and cervico branchial syndrome. There are various analytical methods for estimation of drug. This paper list out the various analytical methods for the drug.

Abstract C070: Investigation of the roles of BDKRB1 in pancreatic cancer progression and tumor microenvironment modulation

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with features of early metastasis and extensive desmoplasia which is the growth of fibrous or connective tissue. The PDAC patient outcomes have not improved in decades, albeit a plenty of therapies that target PDAC have been tested but failed. It could be due to most research focused on tumor cells, whereas the effect of the stroma and immune cells during tumor progression has been largely ignored. Bradykinin receptor B1 (BDKRB1), one of the cell-surface G protein-coupled receptors plays a crucial role in the physiological process associated with coagulation, vasodilation and promotes fibrotic diseases and inflammation in pathological conditions. Based on the analyzed results of Cancer Genome Atlas (TCGA) database, BDKRB1 mRNA is upregulated in PDAC patients. Of note, BDKRB1 mRNA level positively associates with several activated human pancreatic stellate cells (HPSCs; the main fibroblastic cells of the pancreas)-related genes such as CTGF (connective tissue growth factor), IL6 (interleukin 6), and PDGFB (platelet derived growth factor subunit B) in PDAC patients. Interestingly, the conditioned medium from des-Arg9-bradykinin treated-HPSCs can induce M2-like macrophages polarization which dictate clinically relevant immunosuppression in metastatic tumors and express immunosuppressive cytokines such as IL-10, CCL17 and CCL18, implying that activation of BDKRB1 not only stimulates PDAC growth, but also causes HPSCs activation, desmoplasia and immune suppression, followed by exacerbating tumor progression. Moreover, a chemotherapy gemcitabine remains a cornerstone of PDAC treatment in all stages of PDAC despite of the low therapeutic efficacy, we demonstrated that treatment of gemcitabine increases the expression of BDKRB1 in HPSCs and PDAC, indicating that activation of BDKRB1 could be one of the mechanisms involved in gemcitabine resistance. Here, we hypothesize BDKRB1 activation may orchestrate the tumor microenvironment which promotes PDAC progression by inducing tumor cells survival, HPSCs activation, immune suppression, and may contribute to chemotherapy resistance. The outcomes of this proposal will provide not only a novel cellular and molecular mechanism involved in progression of PDAC, but also highlight a potential therapeutic approach to improve PDAC treatment. Citation Format: Ching-Fei Li, Jie Fu, Heng-Huan Lee, Ying-Nai Wang, Yu-Yi Chu, Paul J. Chiao, Mien-Chie Hung. Investigation of the roles of BDKRB1 in pancreatic cancer progression and tumor microenvironment modulation [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr C070.

HEALTHCARE RESOURCE UTILIZATION AMONG LANADELUMAB AND SUBCUTANEOUS C1-INHIBITOR CONCENTRATE NEW USERS - RETROSPECTIVE DATABASE STUDY

<h3>Introduction</h3> Three long-term prophylactic therapies (LTPs) have become available in the past five years for hereditary angioedema (HAE) due to C1-inhibitor (C1-INH) deficiency, including two subcutaneous (SC) options: a monoclonal antibody (lanadelumab) and a plasma-derived C1-INH concentrate (SC-C1-INH). However, limited real-world data on these therapies have been reported. <h3>Methods</h3> Annualized healthcare resource utilization (HCRU), including medication use, and costs of SC-LTP are described among adults starting lanadelumab or SC-C1-INH based on the 180 days before (baseline) and up to 365 days of continuous use after (follow-up) starting therapy, from the IBM MarketScan US insurance claims database. The study population was required to continuously use lanadelumab or SC-C1-INH for=180 days to be in the analysis. <h3>Results</h3> 47 lanadelumab and 38 SC-C1-INH subjects were identified. Bradykinin receptor antagonists (∼50% in both cohorts) and intravenous-C1-INH (40% and 58%, respectively) were the most frequent HAE medications filled during baseline. 20–30% of each cohort had no baseline HAE medication fills. Annualized HCRU, including but not limited to acute medication fills and angioedema-associated emergency department (ED) visits/hospitalizations, decreased by >50% between baseline and follow-up (∼10 to 3–4 for acute medication fills; 1–2 to ∼0.5 for ED visits/hospitalizations) [Figure]. Total estimated annualized healthcare costs were $866,639 and $734,460 for the lanadelumab and SC-C1-INH cohorts respectively, with pharmacy costs accounting for >95% of total costs. <h3>Conclusion</h3> HAE SC-LTP reduced HCRU by >50%, though did not eliminate prescription fills for acute HAE medication nor angioedema-associated ED/hospital utilization, indicating burden of disease persists despite newer treatments.

A CASE OF DIFFICULT TO DIAGNOSE C1INH NORMAL HAE

<h3>Introduction</h3> Hereditary Angioedema (HAE) is characterized by recurrent swellings of the face, tongue, oropharynx, extremities, and abdomen. Typically beginning in the second decade of life, these swells often require emergent treatment and may result in intubation. Unchecked production of bradykinin, typically due to low levels of C1 esterase inhibitor (C1INH), results in bradykinin mediated vascular leakage and swelling. Type I and II can be diagnosed by checking C1INH levels and function but C1INH Normal continues to be difficult to diagnose. We present a challenging case detailing the diagnosis and management of probable C1INH normal HAE in a 42-year-old female. <h3>Case Description</h3> A 42-year-old woman with a history of hypothyroidism, GERD, migraine, obesity, and borderline personality disorder, presented to our clinic with a 10-year history of three or more weekly episodes of throat and eyelid angioedema, accompanied by intermittent urticaria. Her controller therapy included montelukast, fexofenadine, cetirizine, zyflo, famotidine, and cromolyn. Despite this regimen, she continued her thrice weekly episodes, so cyclosporine was added without improvement in flare frequency or severity. C1INH level and function, along with her physical exam, were unremarkable. A trial of recombinant C1INH resolved an acute swell, so she was started on C1INH replacement, with significant reduction in her swell frequency. <h3>Discussion</h3> C1INH normal HAE continues to be difficult to diagnose with currently available assays. For patients with recurrent swells, not responding to mast cell stabilizers or antihistamines, a trial of recombinant C1INH or a bradykinin receptor antagonist, may aid in the diagnosis of C1INH Normal HAE.

KLK12 Regulates MMP-1 and MMP-9 via Bradykinin Receptors: Biomarkers for Differentiating Latent and Active Bovine Tuberculosis.

It has been established that kallikrein12 (KLK12) expression is closely related to bovine tuberculosis (bTB) development. Herein, we sought to clarify the regulatory mechanism of KLK12 and its application in tuberculosis diagnosis. KLK12 knockdown macrophages were produced by siRNA transfection. Bradykinin receptors (BR, including B1R and B2R) were blocked with specific inhibitors. Mannose-capped lipoarabinomannan (ManLAM) was extracted from Mycobacterium bovis (M. bovis) and used to study the mechanism of KLK12 activation. In addition, we constructed different mouse models representing the latent and active stages of M. bovis infection. Mouse models and clinical serum samples were used to assess the diagnostic value of biomarkers. Through the above methods, we confirmed that KLK12 regulates MMP-1 and MMP-9 via BR. KLK12 upregulation is mediated by the M. bovis-specific antigen ManLAM. KLK12, MMP-1, and MMP-9 harbor significant value as serological markers for differentiating between latent and active bTB, especially KLK12. In conclusion, we identified a novel signaling pathway, KLK12/BR/ERK/MMPs, in M. bovis-infected macrophages, which is activated by ManLAM. From this signaling pathway, KLK12 can be used as a serological marker to differentiate between latent and active bTB. Importantly, KLK12 also has enormous potential for the clinical diagnosis of human tuberculosis (TB).

Bradykinin/bradykinin 1 receptor promotes brain microvascular endothelial cell permeability and proinflammatory cytokine release by downregulating Wnt3a.

Stroke is a life-threatening disease with limited therapeutic options. Damage to the blood-brain barrier (BBB) is the key pathological feature of ischemic stroke. This study explored the role of the bradykinin (BK)/bradykinin 1 receptor (B1R) and its mechanism of action in the BBB. Human brain microvascular endothelial cells (BMECs) were used to test for cellular responses to BK by using the Cell Counting Kit-8 assay, 5-ethynyl-2'-deoxyuridine staining, enzyme-linked immunosorbent assay, flow cytometry, immunofluorescence, cellular permeability assays, and western blotting to evaluate cell viability, cytokine production, and reactive oxygen species (ROS) levels in vitro. A BBB induced by middle cerebral artery occlusion was used to evaluate BBB injuries, and the role played by BK/B1R in ischemic/reperfusion (I/R) was explored in a rat model. Results showed that BK reduced the viability of BMECs and increased the levels of proinflammatory cytokines (interleukin 6[IL-6], IL-18, and monocyte chemoattractant protein-1) and ROS. Additionally, cellular permeability was increased by BK treatment, and the expression of tight junction proteins (claudin-5 and occludin) was decreased. Interestingly, Wnt3a expression was inhibited by BK and exogenous Wnt3a restored the effects of BK on BMECs. In an in vivo I/R rat model, knockdown of B1R significantly decreased infarct volume and inflammation in I/R rats. Our results suggest that BK might be a key inducer of BBB injury and B1R knockdown might provide a beneficial effect by upregulating Wnt3a.

ASSOCIATION BETWEEN SPORT RELATED POLYGENIC PROFILE AND MAGNITUDE OF PHYSICAL PERFORMANCES CHANGES FOLLOWING RESISTANCE TRAINING AMONG NOVICE FIELD HOCKEY ATHLETES

Background: Studies have shown that inter-individual variation in response to resistance training is attributed to genetic variation. Aim: To correlate total genotype scores (TGS) with the magnitude of physical performances (skeletal muscle mass, muscular strength, muscular power, performance decrement, and VO2max) change, following 8 weeks of resistance training. Methods: We included forty-five (N=45) participants (age = 16.53 ± .5 years old, body mass = 61.03 ± 6.67 kg, height = 1.67±.05 m) and randomly assigned into; high intensity resistance training (n=15), moderate intensity resistance training (n=15) and a control group (n=15). They were prescribed with the same upper and lower body exercise routines. Pre- and post-test physical performances were measured. Genotyping was conducted using in-house designed allele-specific polymerase chain reaction assays. TGS for nine SNPs: Angiotensin Converting Enzyme, ACE rs1799752; Alpha-acitin-3, ACTN3 rs1815739; Adrenergic Receptor B3, ADRB3 rs4994; Angiotensinogen, AGT rs699; Bradykinin Receptor B2, BDKRB2 rs1799722; Peroxisome Proliferator-Activated Receptor Alpha, PPARA rs4253778; Peroxisome Proliferator-Activated Receptor Gamma Co-Activator 1-Alpha, PPARGC1A rs8192678; Thyrotropin Releasing Hormone Receptor, TRHR rs7832552 and Vascular Endothelial Growth Factor, VEGF rs1870377,were calculated and assigned with strength-power and endurance quality. Pearson correlation analysis was employed to investigate the correlation between TGS, and magnitude of physical performances change following intervention. Result: There is a significant correlation between strength-power TGS with lower body muscular strength (r= .65, p&lt;.01) and power (r= -.69, p&lt;.01) following moderate intensity resistance training. Conclusion: In this study, it is demonstrated that participants with higher strength-power TGS, gained greater lower body strength improvement and lesser muscular power decrement even with moderate intensity as compared to high intensity resistance training. Therefore, personalising training based on athlete’s genetic profile may optimise performance.

Function and structure of bradykinin receptor 2 for drug discovery.

Type 2 bradykinin receptor (B2R) is an essential G protein-coupled receptor (GPCR) that regulates the cardiovascular system as a vasodepressor. Dysfunction of B2R is also closely related to cancers and hereditary angioedema (HAE). Although several B2R agonists and antagonists have been developed, icatibant is the only B2R antagonist clinically used for treating HAE. The recently determined structures of B2R have provided molecular insights into the functions and regulation of B2R, which shed light on structure-based drug design for the treatment of B2R-related diseases. In this review, we summarize the structure and function of B2R in relation to drug discovery and discuss future research directions to elucidate the remaining unknown functions of B2R dimerization.

The Role of Bradykinin Receptors in Hereditary Angioedema Due to C1-Inhibitor Deficiency.

Background: Hereditary angioedema (HAE) is a rare, genetic disease caused by the decreased level or function of the C1 inhibitor. The primary mediator of symptoms in HAE is bradykinin acting through its two receptors, namely receptors 1 (BR1) and 2 (BR2). Although BR2 is well characterized, the role of BR1 remains unclear. Objective: To study the role of bradykinin receptors 1 (BR1) in the etiopathogenesis of HAE. Methods: A total of 70 individuals, 40 patients with HAE, and 30 healthy subjects were recruited to the study. HAE was diagnosed in accordance with the international guideline. The level of bradykinin receptors was determined in populations of CD3+, CD4+, CD8+, and CD14++CD16−, CD14++CD16+ monocytes. In addition, the level of disease activity-specific markers was measured. Results: There were statistically significant differences in the subpopulation of lymphocytes and monocytes between patients with HAE compared to healthy subjects. The level of BR1 and BR2 on PBMCs was comparable in healthy subjects and HAE patients during remission with significant overexpression of both receptors, triggered by HAE attack. Moreover, a significant increase in TNF-alpha and IL-1 plasma levels was observed among HAE patients. Conclusions: BR1 expression may play an important role in the pathomechanism of HAE.

Bioinformatics analysis of the circRNA-miRNA-mRNA network for atrial fibrillation.

Atrial fibrillation (AF) is a chronic and progressive disease, with advancing age, the morbidity of which will increase exponentially. Circular ribonucleic acids (RNAs; circRNAs) have gained a growing attention in the development of AF in recent years. The purpose of this study is to explore the mechanism of circRNA regulation in AF, in particular, the intricate interactions among circRNA, microRNA (miRNA), and messenger RNA (mRNA). Three datasets (GSE129409, GSE68475, and GSE79768) were obtained from the Gene Expression Omnibus database to screen differentially expressed (DE) circRNAs, DE miRNAs, and DE mRNAs in AF, respectively. Based on circRNA-miRNA pairs and miRNA-mRNA pairs, a competing endogenous RNAs (ceRNAs) network was built. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis of DE mRNAs in the network were performed and protein-protein interaction (PPI) networks were established to identify hub genes. Finally, a circRNA-miRNA-hub gene subnetwork was constructed. A total of 103 DE circRNAs, 16 DE miRNAs, and 110 DE mRNAs were screened in AF. Next, ceRNAs network in AF was constructed with 3 upregulated circRNAs, 2 downregulated circRNAs, 2 upregulated miRNAs, 2 downregulated miRNAs, 17 upregulated mRNAs, and 24 downregulated mRNAs. Thirty GO terms and 6 KEGG pathways were obtained. Besides, 6 hub genes (C-X-C chemokine receptor type 4 [CXCR4], C-X-C chemokine receptor type 2 [CXCR2], C-X-C motif chemokine 11 [CXCL11], neuromedin-U, B1 bradykinin receptor, and complement C3) were screened from constructing a PPI network. Finally, a circRNA-miRNA-hub gene subnetwork with 10 regulatory axes was constructed to describe the interactions among the differential circRNAs, miRNA, and hub genes. We speculated that hsa_circRNA_0056281/hsa_circRNA_0006665 -hsa-miR-613-CXCR4/CXCR2/CXCL11 regulatory axes and hsa_circRNA_0003638-hsa-miR-1207-3p-CXCR4 regulatory axis may be associated with the pathogenesis of AF.