A2A and A3, Adenosine receptors mRNA are overexpressed in an experimental animal model of myocardial infarction

Background: Adenosine, a purine nucleoside and a "retaliatory metabolite" in ischemia, is ubiquitous in the body, and increases 100-fold during ischemia. Its biological actions are mediated by four adenosine receptors (ARs): A1 and A3, coupled to Gi/o, and the high-affinity A2A and low-affinity A2B, coupled to Gs. Because A1R and A3R are distributed mainly in myocardial cells and A2 are on coronary vascular smooth cells in the heart, adenosine may substantially modulate cardiac function as a whole. Aim: To determine possible myocardial alterations in the expression of ARs, in an experimental animal model of myocardial infarction (MI). Materials and Methods: Left ventricular (LV) tissue was collected from male adult minipigs with MI (n=5), induced by permanent surgical legation of the left anterior descending coronary artery and from 5 healthy pigs. mRNA expression of A1R, A2AR, A2BR,A3R was determined by semi-quantitative RT-PCR in tissue sampled collected from border (BZ) and remote zones (RZ) of infarcted area. Results: Transmural infarction affected about 15% of the LV wall mass. After 4 weeks, mRNA expression was higher in infarct regions than in control for A1R (controls=2.0?1.0, BZ=2.4?0.4, RZ=1.2?0.1), A2AR (controls=0.6?0.3, BZ=1.9?0.2, RZ=1.3?0.04 p=0.002, p=0.04, controls vs. BZ and RZ), A2BR (controls=1.1?0.5, BZ=1.2?0.2, RZ=0.5?0.04) and A3R (controls=0.2?0.07, BZ=2.4?0.7, RZ=0.7?0.07, p=0.006, p=0.002, controls vs. BZ and RZ). Conclusion: All adenosine receptors, and expecially A2A and A3, are overexpressed in the BZ of MI, consistently with an adaptative retaliatory anti-ischemic adenosinergic changes of post-infarcted heart

Myocardial infarction activates the expression of cardiometabolic biomarkers in the heart: study in a swine model.

Purpose. Inflammation, extra-cellular matrix (ECM) remodeling and adipokine system activation represent essential processes of molecular response to cardiac injury. The aim of this study was to evaluate the cardiac expression of biomarkers involved in the inflammation, ECM remodeling and adiponectin system in an experimental animal model of myocardial infarction (AMI). Methods. Left ventricular (LV) tissue was collected from male adult pigs with AMI (n=6), induced by permanent surgical ligation of the left anterior descending coronary artery and from 6 healthy pigs. mRNA expression was determined by RT-PCR in tissue samples collected from border (BZ) and remote zones (RZ) of infarcted area. Proinflammatory cytokines (IL-6, TNF-&#945;), matrix metalloproteinases (MMP)-2, -9, their tissue inhibitors (TIMP)-1, -2 and collagen (COL3&#945;) were evaluated. In addition, adiponectin and its receptors, adipo-R1 and adipo-R2, were evaluated, owing its anti-inflammatory actions. Results. This surgical approach resulted in a permanent transmural infarction affecting 10-15% of the LV wall mass and after 4 weeks the mRNA expression of biomarkers, normalized on the respective GAPDH, was significantly higher in infarcted regions than in controls (MMP-9: 7.09?4.31; 1.18?0.28; 0.72?0.11, respectively for BZ, RZ and controls, p<0.05 BZ vs. RZ and controls; TIMP-1: 2.41?1.20; 0.28?0.04; 0.33?0.05, p=0.01; TIMP-2: 2.75?1.51; 0.53?0.04; 0.38?0.03, p<0.05; COL3&#945;: 4.28?1.11; 0.87?0.13; 0.61?0.18, p<0.0004). Inflammatory indices were increased in AMI, both BZ and RZ. Adiponectin was significantly increased with respect to controls (BZ: 2.95?1.69; RZ: 0.93?0.33; controls: 0.52?0.12, p<0.05 BZ vs controls) as well as the Adipo-R1 (BZ: 1.40?0.31, RZ: 1.26?0.20, controls: 0.63?0.07; p<0.05 BZ and RZ vs controls). Conclusions. The inflammatory and ECM remodelling processes are activated after myocardial injury together with the system of adiponectin, confirming its involvement in the process of cardiac remodelling/repair. The knowledge of the interaction between the various mediators of the complex response to cardiac damage could represent an important target for new therapies. Reference. Shibata R et al, Cardiovasc Res. 2007 Jun 1;74(3):471-9

Mismatch between mRNA cardiac expression of BNP and CNP in pacing-induced heart failure

Purpose: It has been recently demonstrated in an animal model of heart failure (HF) that the high-frequency pacing of the left ventricle (LV) free wall causes a dyssynchronous pattern of contraction that leads to progressive cardiac failure with pronounced differences in regional contractility. Aim of this study was to evaluate the possible variation of brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) mRNA expression in the anterior/anterior lateral region (pacing site, PS) compared with the infero-septal region (opposite site, OS) to individualize the possible association between the contraction patterns and the expression of these biomarkers. Methods: Cardiac tissue was collected from male adult minipigs without (controls, n=6) and with pacing-induced HF (n=8) from PS, and from the tissue remote from the pacing-site. mRNA expression of BNP and CNP was evaluated by semiquantitative polymerase chain reaction (PCR) by using GAPDH as housing gene. Results: A significant difference in mRNA expression of BNP in PS was observed between HF animals and controls (BNP/GAPDH: 0.65?0.11 vs. 0.35?0.04, p=0.02) whereas in OS BNP levels resulted similar to those of controls (BNP/GAPDH: 0.36?0.05). mRNA expression of CNP was higher in HF with respect to controls both in PS and in OS, although not significantly (CNP/GAPDH: controls 0.089?0.036, PS 0.29?0.23, OS 0.54?0.16). These findings are in tune with the increase of CNP tissue concentrations (controls=0.69?0.13; PS=1.56?0.19; OS=1.70?0.42 pg/mg protein; p=0.039 controls vs.OS). The higher levels of BNP mRNA expression in PS are in agreement with a reduction of contractile function in this region while the higher CNP mRNA expression in OS could suggest the presence of a major endothelial dysfunction in the remote region. Conclusions: In clinical conditions the endothelial dysfunction precedes the overt HF, so, although further investigations are necessary, these results suggest that CNP could be a early marker of HF. In this context, CNP could be a marker more relevant than BNP in early recognizing patients with HF

Adenosine receptor expression in an experimental animal model of myocardial infarction with preserved left ventricular ejection fraction.

Adenosine, a purine nucleoside and a "retaliatory metabolite" in ischemia, is ubiquitous in the body and increases 100-fold during ischemia. Its biological actions are mediated by four adenosine receptors (ARs): A(1), A(2A), A(2B) and A(3). The aim of this study was to determine possible myocardial alterations in AR expression in an experimental animal model of myocardial infarction (MI) with a preserved left ventricular (LV) ejection fraction. LV tissue was collected from sexually mature male farm pigs with MI (n = 6) induced by permanent surgical ligation of the left anterior descending coronary artery and from five healthy pigs (C). mRNA expression of A(1)R, A(2A)R, A(2B)R, A(3)R and TNF-? was determined by real-time PCR in tissue collected from border (BZ) and remote zones (RZ) of the infarcted area and from LV of C. BZ, RZ and samples of C were stained immunohistochemically to investigate A(3)R immunoreaction. After 4 weeks a different regulation of ARs was observed. A(1)R mRNA expression was significantly lower in the infarct regions than in controls (C = 0.75 ? 0.2, BZ = 0.05 ? 0.2, RZ = 0.07 ? 0.02 p = 0.0025, p = 0.0016, C vs. BZ and RZ, respectively). Conversely A(3)R was higher in infarct areas (C = 0.94 ? 0.2, BZ = 2.85 ? 0.5, RZ = 3.48 ? 1.0, p = 0.048 C vs. RZ). No significant differences were observed for A(2A)R (C = 1.58 ? 0.6, BZ = 0.42 ? 0.1, RZ = 1.37 ? 0.6) and A(2B)R (C = 1.66 ? 0.2, BZ = 1.54 ? 0.5, RZ = 1.25 ? 0.4). A(3)R expression was confirmed by immunohistochemical analysis and was principally localized in cardiomyocytes. TNF-? mRNA results were: C 0.41 ? 0.25; BZ 1.60 ? 0.19; RZ 0.17 ? 0.04. The balance between A(1)R and A(3)R as well as between A(2A)R and A(2B)R was consistent with adaptative retaliatory anti-ischemic adenosinergic changes in the infarcted heart with preserved LV function

Homo sapiens natriuretic peptide precursor type C (NPPC) mRNA,partial cds and 3\u27 UTR.

LOCUS HQ419060 318 bp mRNA linear PRI 24-NOV-2010 DEFINITION Homo sapiens natriuretic peptide precursor type C (NPPC) mRNA, partial cds and 3\u27 UTR. ACCESSION HQ419060 VERSION HQ419060.1 GI:312261407 KEYWORDS . SOURCE Homo sapiens (human) ORGANISM Homo sapiens Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominidae; Homo. REFERENCE 1 (bases 1 to 318) AUTHORS Landi,S., Melaiu,O., Cabiati,M., Landi,D., Caselli,C., Prescimone,T., Giannessi,D., Gemignani,F. and Del Ry,S. TITLE Direct Submission JOURNAL Submitted (20-OCT-2010) Laboratory of Cardiovascular Biochemistry, Institute of Clinical Physiology, Via Moruzzi 1, Pisa, PI 56100, Italy FEATURES Location/Qualifiers source 1..318 /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /cell_line="SKNBE" /PCR_primers="fwd_seq: gtcagaagaagggcgacaag, rev_seq: gcgtttaaacgcgcacgcgt" gene <1..318 /gene="NPPC" CDS <1..188 /gene="NPPC" /codon_start=3 /product="natriuretic peptide precursor type C" /protein_id="ADQ54381.1" /db_xref="GI:312261408" /translation="GDRSRLLRDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKG CFGLKLDRIGSMSGLGC" 3\u27UTR 189..318 /gene="NPPC" ORIGIN 1 agggcgaccg gtcgcgactg ctccgggacc tgcgcgtgga caccaagtcg cgggcagcgt 61 gggctcgcct tctgcaagag caccccaacg cgcgcaaata caaaggagcc aacaagaagg 121 gcttgtccaa gggctgcttc ggcctcaagc tggaccgaat cggctccatg agcggcctgg 181 gatgttagtg cggcgccccc tggcggcggg agaagaatga ttctgacact tggggaccag 241 ccttcagtag ctacccttgg aatgcctttg ctctcttctc tcctgtctaa acaacaaaga 301 gacggagtct gaggcct

Adiponectin and cardiometabolic risk factor: effect on myocardial blood flow in patients with dilated cardiomyopathy

Purpose: Adiponectin (ADN) is an insulin-sensitizing, anti-atherogenic and anti-inflammatory adipocytokine, with endothelial protective effects. In patients with dilated cardiomyopathy (DCM), absolute myocardial blood flow (MBF) is frequently impaired because of coronary microvascular/endothelial dysfunction. We aimed at evaluating whether the potential effects of ADN on MBF in DCM are modulated by cardiometabolic risk factors. Methods: Sixty-one consecutive patients (46 males, age 59?10 yrs) with LV dysfunction (LV ejection fraction 38?9%, range 19-54%) and angiographically normal coronary arteries were studied. Absolute MBF was measured by positron emission tomography, using 13N-Ammonia as flow tracer, both at rest and during dipyridamole infusion (0.56 mg/kg I.V. over 4 min). Abnormal MBF was defined for resting MBF<0.65 ml/min/g and dipyridamole MBF<1.36 ml/min/g. Plasma adiponectin was measured by a specific ELISA (Linco Research). Cardiometabolic risk factors including age, sex, insulin resistance, lipid profile and obesity were entered into a multivariate linear regression analysis to assess independent determinants of adiponectin and, in turn, their effect on myocardial and microvascular damage. Results: Abnormal MBF was present in 34/61 patients, in 13/31 (42%) with ADN higher than median value and in 21/30 (70%) with ADN levels lower than median value (p<0.05). Among cardiometabolic risk factors, only HDL-cholesterol and obesity were independent determinants of ADN [0.34 (0.13), -0.34 (0.14), &#946;-coefficient (SE), respectively, p<0.05] at multivariate analysis. After correction for HDL-cholesterol and obesity the positive association between ADN and MBF remained significant (p<0.05). Conclusion: ADN is a determinant of MBF in DCM patients and is related with low HDL-cholesterol and obesity

High concentration of C-type natriuretic peptide promotes VEGF-dependent vasculogenesis in the remodeled region of infarcted swine heart with preserved left ventricular ejection fraction.

Abstract BACKGROUND: Vasculogenesis is a hallmark of myocardial restoration. Post-ischemic late remodeling is associated with pathology and function worsening. At the same time, neo-vasculogenesis helps function improving and requires the release of vascular endothelial growth factor type A (VEGF-A). The vasculogenic role of C-type natriuretic peptide (CNP), a cardiac paracrine hormone, is unknown in infarcted hearts with preserved left ventricular (LV) ejection fraction (EF). We explored whether myocardial VEGF-dependent vasculogenesis is affected by CNP. METHODS AND RESULTS: To this end, infarcted swine hearts were investigated by magnetic resonance imaging (MRI), histological and molecular assays. At the fourth week, MRI showed that transmural myocardial infarction (MI) affected approximately 13% of the LV wall mass without impairing global function (LVEF>50%, n=9). Increased fibrosis, metalloproteases and capillary density were localized to the infarct border zone (BZ), and were associated with increased expression of CNP (p=0.03 vs. remote zone (RZ)), VEGF-A (p<0.001 vs. RZ), BNP, a marker of myocardial dysfunction (p<0.01 vs. RZ) and the endothelial marker, factor VIII-related antigen (p<0.01 vs. RZ). In vitro, CNP 1000 nM promoted VEGF-dependent vasculogenesis without affecting the cell growth and survival, although CNP 100 nM or a high concentration of VEGF-A halted vascular growth. CONCLUSIONS: CNP expression is locally increased in infarct remodeled myocardium in the presence of dense capillary network. The vasculogenic response requires the co-exposure to high concentration of CNP and VEGF-A. Our data will be helpful to develop combined myocardial delivery of CNP and VEGF-A genes in order to reverse the remodeling process

ADIPONECTIN AND CARDIOVASCULAR RISK PREDICTION: STRATIFICATION OF CHEST PAIN PATIENTS BY A CLUSTER ANALYSIS

Cardiovascular disease (CVD) remains the major cause of death and there is the need to a better stratification of CVD patients. By an unbiased statistical approach we sought to identify clusters of patients to better stratify their risk. 202 patients with chest pain (63% males, age 62?12 yr) undergone to CT coronary angiography (CCTA) were prospectively included and classified using K-means cluster analysis of clinical, imaging and bio-humoral data. The most relevant classification resulted in three phenotypes distinguished according to Framingham score and HMW adiponectin plasma levels. Presence and severity of disease as assessed by CCTA were verified trough these phenotypes. By K-means cluster analysis, we identified CVD phenotypes allowing to stratify patients requiring different diagnostic and therapeutic approach

Livelli plasmatici di adrenomedullina come indicatori di prognosi dopo terapia di resincronizzazione cardiaca

The cardiac resynchronization therapy (CRT), based on correction of electro-mechanical dyssynchrony by biventricular pacing in patientswith severe chronicHF unresponsive to optimalmedical treatment and left ventricular conduction disturbances, has been developed. The determination of plasma adrenomedullin (ADM) levels before implantation could provide important additional information to reduce the high percentage (30%) of patients not responding to treatment despite the use of increasingly sophisticatedmethods for selecting candidates. The case described illustrates the importance of basal ADM plasma levels in predicting the clinical and functional improvement after treatment with CRT

Exploring PTX3 expression in Sus scrofa cardiac tissue using RNA sequencing

The prototypic long pentraxin PTX3 is a novel vascular inflammatory marker sharing similarities with the classic short pentraxin (C-reactive protein). PTX3 is rapidly produced and released by several cell types in response to local inflammation of the cardiovascular system. Plasma PTX3 levels are very low in normal conditions and increase in heart failure (HF) patients with advancing NYHA functional class, but its exact role during HF pathogenetic mechanisms is not yet established. No data about PTX3 cardiac expression in normal and pathological conditions are currently available, either in human or in large-size animals. Of the latter, the pig has a central role in "in vivo" clinical settings but its genome has not been completely sequenced and the PTX3 gene sequence is still lacking. The aim of this study was to sequence the PTX3 in Sus scrofa, whose sequence is not yet present in GenBank. Utilizing our knowledge of this sequence, PTX3 mRNA expression was evaluated in cardiac tissue of normal (n=6) and HF pigs (n=5), obtained from the four chambers. To sequence PTX3 gene in S. scrofa, the high homology between Homo sapiens and S. scrofa was exploited. Pig PTX3 mRNA was sequenced using polymerase chain reaction primers designed from human consensus sequences. The DNA, obtained from different RT-PCR reactions, was sequenced using the Sanger method. S. scrofa PTX3 mRNA, 1-336bp, was submitted to GenBank (ID: GQ412351). The sequence obtained from pig cardiac tissue shared an 84% sequence identity with human homolog. The presence of PTX3 mRNA expression was detected in all the cardiac chambers sharing an increase after 3weeks of pacing compared to controls (p=0.036 HF right atrium vs. N; p=0.022, HF left ventricle vs. N). Knowledge of the PTX3 sequence could be a useful starting point for future studies devoted to better understanding the specific role of this molecule in the pathogenesis of cardiovascular disease