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Corin is down-regulated and exerts cardioprotective action via activating pro-atrial natriuretic peptide pathway in diabetic cardiomyopathy.

Pang A, Hu Y, Zhou P, Long G, Tian X, Men L, Shen Y, Liu Y, Cui Y - Cardiovasc Diabetol (2015)

Bottom Line: The effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration was measured by the wound healing scratch assay.The corin and ANP expression in mRNA and protein levels was decreased in DCM rat hearts.Corin-siRNA H9c2 cardiomyoblasts showed decreased proliferation.

View Article: PubMed Central - PubMed

Affiliation: Hematopoietic Stem Cell Transplantation Center, Institute of Hematology and Blood Diseases Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin, 300020, China. aimingpangdoc@live.cn.

ABSTRACT

Background: Diabetic cardiomyopathy (DCM), a fatal cardiovascular complication of diabetes mellitus, often leads to progressive heart failure, however its pathogenesis remains unclear. Corin, a cardiac serine protease, is responsible for converting pro-atrial natriuretic peptide (pro-ANP) to biologically active atrial natriuretic peptide (ANP). It has been well established that corin deficiency is associated with the progression of hypertension, cardiac hypertrophy and heart failure. However, because the involvement of corin-mediated pro-ANP processing in DCM has not been clarified, this study aims to investigate the role of corin in the pathogenesis of DCM.

Methods: Diabetes mellitus was induced by a single intraperitoneal injection of streptozotocin (STZ 65 mg/kg) to Sprague-Dawley rats (180-220 g). DCM was confirmed by monitoring continuously transthoracic echocardiography every 4 weeks and hemodynamic measurements at 20 weeks. Myocardial disorder and fibrosis were detected by HE staining and Masson's trichrome staining. The mRNA and protein levels of corin and ANP in rat hearts and cardiomyocytes were determined by quantitative real-time PCR, western blotting and immunohistochemical staining, respectively. H9c2 cardiomyoblasts proliferation was detected by MTT colorimetric assay and viable cell counting with trypan blue. The effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration was measured by the wound healing scratch assay.

Results: The corin and ANP expression in mRNA and protein levels was decreased in DCM rat hearts. Corin and ANP levels of neonatal rat cardiomyocytes and H9c2 cardiomyoblasts treated with high glucose were significantly lower than that of normal glucose treated. Precisely, corin and ANP levels decreased in DCM rats at 12, 16, 20 and 33 weeks; neonatal cardiomyocytes and H9c2 cardiomyoblasts treated with high glucose at 36, 48 and 60 h demonstrated significant reduction in corin and ANP levels. Corin-siRNA H9c2 cardiomyoblasts showed decreased proliferation. Culture supernatants of Corin-siRNA H9c2 cardiomyoblasts prevented endothelial cell line EA.hy926 migration in the wound healing scratch assay. Furthermore, iso-lectin expression in arteriole and capillary endothelium was down-regulated in DCM rats.

Conclusions: Our results indicate that corin plays an important role in cardioprotection by activating pro-atrial natriuretic peptide pathway in DCM. Corin deficiency leads to endothelial dysfunction and vascular remodeling.

No MeSH data available.


Related in: MedlinePlus

Defect of corin was associated with endothelial dysfunction. a The wound healing scratch assay showed the effect of culture supernatants of H9c2 cardiomyoblasts transfected Corin-siRNA or NC-siRNA on EA.hy926 cells migration at 0 and 24 h. Bar 250 μm. b, c Reduced capillary and arteriole densities in DCM rat myocardium were detected by confocal fluorescence microscope. Capillary and arteriole densities were stained by iso-lectin B4 (green) in rat hearts, nuclei were stained with Dapi (blue). Bar 50 μm (n = 5 to 6 rats for each group). For quantification, 4 or 5 randomly selected fields from each rat myocardial tissue under ×400 magnification were analyzed. Data are presented as mean ± SD. *P < 0.05 versus Ctrl
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Fig6: Defect of corin was associated with endothelial dysfunction. a The wound healing scratch assay showed the effect of culture supernatants of H9c2 cardiomyoblasts transfected Corin-siRNA or NC-siRNA on EA.hy926 cells migration at 0 and 24 h. Bar 250 μm. b, c Reduced capillary and arteriole densities in DCM rat myocardium were detected by confocal fluorescence microscope. Capillary and arteriole densities were stained by iso-lectin B4 (green) in rat hearts, nuclei were stained with Dapi (blue). Bar 50 μm (n = 5 to 6 rats for each group). For quantification, 4 or 5 randomly selected fields from each rat myocardial tissue under ×400 magnification were analyzed. Data are presented as mean ± SD. *P < 0.05 versus Ctrl

Mentions: To determine the relationship between corin expression and endothelial function, we further detected the effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration. The migratory speed of EA.hy926 cells treated with culture supernatants of Corin-siRNA H9c2 cardiomyoblasts was markedly decreased compared with that in NC-siRNA group using the wound healing assay (Fig. 6a). These results demonstrated the ability of corin to protect against endothelial dysfunction in vitro. Furthermore, iso-lectin B4 expression in DCM rats was lower than that in the control group (Fig. 6b, c). These findings suggested that the lack of corin impaired endothelial function and ultimately led to vascular remodeling.Fig. 6


Corin is down-regulated and exerts cardioprotective action via activating pro-atrial natriuretic peptide pathway in diabetic cardiomyopathy.

Pang A, Hu Y, Zhou P, Long G, Tian X, Men L, Shen Y, Liu Y, Cui Y - Cardiovasc Diabetol (2015)

Defect of corin was associated with endothelial dysfunction. a The wound healing scratch assay showed the effect of culture supernatants of H9c2 cardiomyoblasts transfected Corin-siRNA or NC-siRNA on EA.hy926 cells migration at 0 and 24 h. Bar 250 μm. b, c Reduced capillary and arteriole densities in DCM rat myocardium were detected by confocal fluorescence microscope. Capillary and arteriole densities were stained by iso-lectin B4 (green) in rat hearts, nuclei were stained with Dapi (blue). Bar 50 μm (n = 5 to 6 rats for each group). For quantification, 4 or 5 randomly selected fields from each rat myocardial tissue under ×400 magnification were analyzed. Data are presented as mean ± SD. *P < 0.05 versus Ctrl
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4597453&req=5

Fig6: Defect of corin was associated with endothelial dysfunction. a The wound healing scratch assay showed the effect of culture supernatants of H9c2 cardiomyoblasts transfected Corin-siRNA or NC-siRNA on EA.hy926 cells migration at 0 and 24 h. Bar 250 μm. b, c Reduced capillary and arteriole densities in DCM rat myocardium were detected by confocal fluorescence microscope. Capillary and arteriole densities were stained by iso-lectin B4 (green) in rat hearts, nuclei were stained with Dapi (blue). Bar 50 μm (n = 5 to 6 rats for each group). For quantification, 4 or 5 randomly selected fields from each rat myocardial tissue under ×400 magnification were analyzed. Data are presented as mean ± SD. *P < 0.05 versus Ctrl
Mentions: To determine the relationship between corin expression and endothelial function, we further detected the effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration. The migratory speed of EA.hy926 cells treated with culture supernatants of Corin-siRNA H9c2 cardiomyoblasts was markedly decreased compared with that in NC-siRNA group using the wound healing assay (Fig. 6a). These results demonstrated the ability of corin to protect against endothelial dysfunction in vitro. Furthermore, iso-lectin B4 expression in DCM rats was lower than that in the control group (Fig. 6b, c). These findings suggested that the lack of corin impaired endothelial function and ultimately led to vascular remodeling.Fig. 6

Bottom Line: The effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration was measured by the wound healing scratch assay.The corin and ANP expression in mRNA and protein levels was decreased in DCM rat hearts.Corin-siRNA H9c2 cardiomyoblasts showed decreased proliferation.

View Article: PubMed Central - PubMed

Affiliation: Hematopoietic Stem Cell Transplantation Center, Institute of Hematology and Blood Diseases Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin, 300020, China. aimingpangdoc@live.cn.

ABSTRACT

Background: Diabetic cardiomyopathy (DCM), a fatal cardiovascular complication of diabetes mellitus, often leads to progressive heart failure, however its pathogenesis remains unclear. Corin, a cardiac serine protease, is responsible for converting pro-atrial natriuretic peptide (pro-ANP) to biologically active atrial natriuretic peptide (ANP). It has been well established that corin deficiency is associated with the progression of hypertension, cardiac hypertrophy and heart failure. However, because the involvement of corin-mediated pro-ANP processing in DCM has not been clarified, this study aims to investigate the role of corin in the pathogenesis of DCM.

Methods: Diabetes mellitus was induced by a single intraperitoneal injection of streptozotocin (STZ 65 mg/kg) to Sprague-Dawley rats (180-220 g). DCM was confirmed by monitoring continuously transthoracic echocardiography every 4 weeks and hemodynamic measurements at 20 weeks. Myocardial disorder and fibrosis were detected by HE staining and Masson's trichrome staining. The mRNA and protein levels of corin and ANP in rat hearts and cardiomyocytes were determined by quantitative real-time PCR, western blotting and immunohistochemical staining, respectively. H9c2 cardiomyoblasts proliferation was detected by MTT colorimetric assay and viable cell counting with trypan blue. The effect of Corin-siRNA H9c2 cardiomyoblasts on EA.hy926 cells migration was measured by the wound healing scratch assay.

Results: The corin and ANP expression in mRNA and protein levels was decreased in DCM rat hearts. Corin and ANP levels of neonatal rat cardiomyocytes and H9c2 cardiomyoblasts treated with high glucose were significantly lower than that of normal glucose treated. Precisely, corin and ANP levels decreased in DCM rats at 12, 16, 20 and 33 weeks; neonatal cardiomyocytes and H9c2 cardiomyoblasts treated with high glucose at 36, 48 and 60 h demonstrated significant reduction in corin and ANP levels. Corin-siRNA H9c2 cardiomyoblasts showed decreased proliferation. Culture supernatants of Corin-siRNA H9c2 cardiomyoblasts prevented endothelial cell line EA.hy926 migration in the wound healing scratch assay. Furthermore, iso-lectin expression in arteriole and capillary endothelium was down-regulated in DCM rats.

Conclusions: Our results indicate that corin plays an important role in cardioprotection by activating pro-atrial natriuretic peptide pathway in DCM. Corin deficiency leads to endothelial dysfunction and vascular remodeling.

No MeSH data available.


Related in: MedlinePlus