Cardioprotective effects of zofenopril, enalapril and valsartan against ischaemia/reperfusion injury as well as doxorubicin cardiotoxicity.

OBJECTIVE: The aim of this study is to compare possible protective effects of zofenopril, enalapril and valsartan against both ischaemia/reperfusion injury as well as acute doxorubicin cardiotoxicity. All three agents have never been compared in this setting before. METHODS AND RESULTS: Sixty-four male rats were divided into eight groups by computer-generated random numbers and each group included 8 rats. Groups 1, 2, 3 and 4, respectively, received 0.5 ml distilled water, 15 mg/kg/day zofenopril, 2 mg/kg/day enalapril, and 30 mg/kg/day valsartan intragastrically for 7 days. Groups 5, 6, 7, and 8 underwent the same procedures as groups 1, 2, 3 and 4. On the 7th day, groups 1-4 and groups 5-8, respectively, were injected with serum saline or 20 mg/kg doxorubicin intraperitoneally. On the 9th day, isolated rat hearts were perfused in the Langendorff perfusion system. At the end of each Langendorff experiment, the rat hearts were kept for histological analysis. Left ventricular systolic pressures were negatively affected by doxorubicin with ischaemia (group 5 initially: 61.4 +/- 13.6 mmHg--post-ischaemic (PI): 20.7 +/- 17.5 mmHg (P = 0.0002), group 6 initially: 63 +/- 18.2 mmHg--PI: 24.2 +/- 24.3 mmHg (P = 0.0135), group 7:82 +/- 26 mmHg--PI: 14.3 +/- 12.1 mmHg (P < 0.0001), group 8:73.1 +/- 27.8 mmHg--PI: 20.4 +/- 27.3 mmHg (P < 0.0001). The lowest troponin I levels (group 2: 0.3 +/- 0.2 ng/ml, group 6:0.2 +/- 0.1 ng/ml (P = 0.003) versus the groups' baseline value) were recorded in the groups of zofenopril in the coronary perfusate during post-ischaemic period. Light microscopic evaluation revealed marked cardiac damage with doxorubicin, since zofenopril treatment prevented a doxorubicin induced increase in the histopathological scores. CONCLUSIONS: In respect of our results zofenopril could be considered more effective than enalapril and valsartan in protecting against both ischaemia/reperfusion injury as well as doxorubicin induced-cardiotoxicity.

Divergent roles of prokineticin receptors in the endothelial cells: angiogenesis and fenestration.

Prokineticins are secreted peptides that activate two G protein-coupled receptors: PKR1 and PKR2. Prokineticins induce angiogenesis and fenestration, but the cognate receptors involved in these functions are unknown. We hypothesized a role for prokineticin receptor signaling pathways and expression profiles in determining the selective effects of prokineticins on coronary endothelial cells (H5V). Activation of the PKR1/MAPK/Akt signaling pathway stimulates proliferation, migration, and angiogenesis in H5V cells, in which PKR1 predominates over PKR2. PKR1 was colocalized with Galpha(11) and was internalized following the stimulation of these cells with prokineticin-2. Knock down of PKR1 or Galpha(11) expression in H5V cells effectively inhibited prokineticin-2-induced vessel formation and MAPK/Akt activation, indicating a role for PKR1/Galpha(11) in this process. However, in conditions in which PKR2 predominated over PKR1, these cells displayed a fenestrated endothelial cell phenotype. H5V cells overexpressing PKR2 displayed large numbers of multivesicular bodies and caveolar clusters and a disruption of the distribution of zonula occluden-1 tight junction protein. Prokineticin-2 induced the colocalization of PKR2 with Galpha(12), and activated Galpha(12), which bound to zonula occluden-1 to trigger the degradation of this protein in these cells. Prokineticin-2 induced the formation of vessel-like structures by human aortic endothelial cells expressing only PKR1, and disorganized the tight junctions in human hepatic sinusoidal endothelial cells expressing only PKR2, confirming the divergent roles of these receptors. Our findings show the functional characteristics of coronary endothelial cells depend on the expression of PKR1 and PKR2 levels and the divergent signaling pathways used by these receptors.

Transgenic myocardial overexpression of prokineticin receptor-2 (GPR73b) induces hypertrophy and capillary vessel leakage.

AIMS: Prokineticins are small secreted bioactive molecules. They exert their biological activity by binding to two G protein-coupled receptors. Previously, we have shown that the overexpression of prokineticin receptor-1 (PKR1) in transgenic (TG) mouse hearts induced neovascularization. Since PKR1 and PKR2 are 85% identical and expressed in cardiovascular tissues, we hypothesized that PKR2 may also contribute to cardiomyocyte growth and vascularization. METHODS AND RESULTS: We have generated TG mice overexpressing PKR2 in cardiomyocytes. TG mice exhibit increased hypertrophic gene expression and heart-to-body weight ratio accompanied by an increased length of cardiomyocytes at the age of 12 weeks. Increased left ventricular end-systolic and diastolic diameters without cardiac dysfunction at the age of 24 weeks indicate that TG mice have an eccentric hypertrophy with compensated cardiac function. Quantitative morphological analysis showed that TG hearts have a normal microvessel density and number of branch points. However, they exhibit increased abnormal endothelial cell shape and ultrastructure, changed cellular distribution of a tight junction protein zona occludens-1 (ZO-1), and vascular leakage in heart without a rise of angiogenic factor levels at early and late age. The application of media conditioned by H9c2 cardioblast cells overexpressing PKR2 significantly induced impaired ZO-1 localization in H5V endothelial cells, mimicking the TG model. CONCLUSION: These findings provide the first genetic evidence that cardiomyocyte PKR2 signalling leads to eccentric hypertrophy in an autocrine regulation and impaired endothelial integrity in a paracrine regulation without inducing angiogenesis. These TG mice may provide a new genetic model for heart diseases.