Expression of Metallopeptidases and Kinin Receptors in Swine Oropharyngeal Tissues: Effects of Angiotensin I-Converting Enzyme Inhibition and Inflammation

Abstract

Angiotensin I-converting enzyme inhibitors (ACEi) cause both chronic and acute side effects, including rare but potentially life-threatening angioedema (AE). The main hypothesis to be tested in this study was that metallopeptidases and kinin receptors are present in oropharyngeal tissues and that their expression is modulated by ACEi and inflammation. Novel realtime polymerase chain reaction analysis was developed and allowed the relative quantification of tissue's gene expression for neprilysin, membrane-bound aminopeptidase P (mAPP), and both B 1 and B 2 kinin receptor subtypes in tongue, parotid gland, and laryngeal tissue (areas especially involved in the gravest clinical forms of AE) and in kidney in a porcine model (single injection or 7-day ACEi oral treatments applied or lipopolysaccharide injected as a positive inflammatory control). The results provide evidence of the expression and activities of kininases in oropharyngeal tissues in the swine. ACEi treatment modulated the expression of neutral endopeptidase and mAPP mRNA, but the corresponding enzyme activities and that of angiotensin I-converting enzyme (ACE) were generally stable in tissues. The 7-day ACEi treatment up-regulated both kinin receptor mRNAs in the oropharynx and the B 1 receptor mRNA in the lingual vascular endothelium (immunohistochemistry). The inhibition of ACE in plasma is responsible for an accumulation of bradykinin and des-arginine 9 -bradykinin generated during activation of the contact system with glass beads. The expression of critical components of the kallikrein-kinin system in the oropharyngeal tissues supports the role of kinins in ACEi-induced AE.

Gene expression analysis of interferon κ in laser capture microdissected cervical epithelium

“…Furthermore, many published techniques claiming high-quality RNA isolation from FR tissue either relied only on expression of HKGs [2,3] and A 260 /A 280 ratios [34,35] or did not perform RNA quality assessment at all [8,10,36]. Here we show that contrary to previous results with FR tissue [4,[37][38][39], the modified Arcturus RNA extraction method was optimal for the extraction of high-quality RNA compared with the Ambion and Sigma methods for cervical biopsy sections (Fig.…”

Role of Nuclear Factor-κB and Protein Kinase C Signaling in the Expression of the Kinin B1Receptor in Human Vascular Smooth Muscle Cells

“…We have recently observed that pigs undergoing long-term treatment with the angiotensin I-converting enzyme inhibitor enalapril exhibit a generalized B 1 receptor up-regulation (mRNA in several organs and endothelial immunoreactivity in tissue sections). This protracted response occurred without changes in the white blood cell counts or increases in the phase reactant C-reactive protein (Moreau et al, 2005a). On the other hand, pharmacological inhibition did not support important roles for tyrosine or MAP kinases in the basal or stimulated expression of B 1 receptors in the present cellular model (data not shown), in contrast to other in vitro systems Phagoo et al, 2001).…”

An in vitro reconstitution system to address the mechanism of the vascular expression of the bradykinin B1 receptor in response to angiotensin converting enzyme inhibition

“…Indeed, B 1 R expression in IMR-90 embryonic fibroblasts has been shown to be acutely stimulated by agonists of either the B 1 or B 2 Rs, but via the release of autocrine inflammatory cytokines (Phagoo et al, 1999). The protein kinase C-dependent and cytokine-independent pathway for B 1 R autoregulation would be compatible with the absence of systemic inflammation in pigs chronically treated with an ACE inhibitor (Moreau et al, 2005a) and with the repressor effect of 18 a B 1 R receptor antagonist on B 1 R mRNA expression in tissues of rats treated with such a drug (Marin-Castano et al, 2002). The lack of effect of all other tested peptides potentially affected by ACE inhibition also does not support that a cross-talk between peptides affected by ACE and macrophages has a role in vascular B 1 R expression.…”

“…Notably, in the porcine model, there was no evidence of systemic inflammation (fever, leukocyte count alteration, acute phase protein release) parallel to B 1 R expression (Moreau et al, 2005a).…”

“…In addition, limited but intriguing evidence has associated B 1 R expression to chronic inhibition of angiotensin I converting enzyme (ACE). There is evidence in 3 animal species that B 1 R expression is triggered in healthy subjects without obvious deleterious effect (Marin-Castano et al, 2002;Moreau et al, 2005a;Bawolak et al, 2008).…”