Inhibition of sodium glucose cotransporter 2 (SGLT2) has been reported as a new therapeutic strategy for treating diabetes. However, the effect of SGLT2 inhibitors on the kidney is unknown. In addition, whether SGLT2 inhibitors have an anti-inflammatory or antioxidative stress effect is still unclear. In this study, to resolve these issues, we evaluated the effects of the SGLT2 inhibitor, dapagliflozin, using a mouse model of type 2 diabetes and cultured proximal tubular epithelial (mProx24) cells. Male db/db mice were administered 0.1 or 1.0 mg/kg of dapagliflozin for 12 weeks. Body weight, blood pressure, blood glucose, hemoglobin A1c, albuminuria and creatinine clearance were measured. Mesangial matrix accumulation and interstitial fibrosis in the kidney and pancreatic β-cell mass were evaluated by histological analysis. Furthermore, gene expression of inflammatory mediators, such as osteopontin, monocyte chemoattractant protein-1 and transforming growth factor-β, was evaluated by quantitative reverse transcriptase-PCR. In addition, oxidative stress was evaluated by dihydroethidium and NADPH oxidase 4 staining. Administration of 0.1 or 1.0 mg/kg of dapagliflozin ameliorated hyperglycemia, β-cell damage and albuminuria in db/db mice. Serum creatinine, creatinine clearance and blood pressure were not affected by administration of dapagliflozin, but glomerular mesangial expansion and interstitial fibrosis were suppressed in a dose-dependent manner. Dapagliflozin treatment markedly decreased macrophage infiltration and the gene expression of inflammation and oxidative stress in the kidney of db/db mice. Moreover, dapagliflozin suppressed the high-glucose-induced gene expression of inflammatory cytokines and oxidative stress in cultured mProx24 cells. These data suggest that dapagliflozin ameliorates diabetic nephropathy by improving hyperglycemia along with inhibiting inflammation and oxidative stress.

How left ventricular (LV) contractility relates to irregular RR intervals during atrial fibrillation (AF) is still unclear. We investigated the relationship between the LV contractility (Emax) of individual beats and their preceding RR intervals during AF in isovolumic contractions is excised cross-circulated canine hearts, and additionally in ejecting contractions in in situ canine hearts. Atrial high-frequency electrical stimulation induced AF. We recorded a LV electrocardiogram, volume and pressure, and calculated the Emax of every arrhythmic beat. Multiple linear regression analysis between Emax and the six preceding RR intervals of all arrhythmic beats during 1 min AF showed the preceding RR interval (RR1) and the pre-preceding interval (RR2) to be the predominant predictors of Emax. The Emax-RR1/RR2 scattergram was closely fitted by a linear regression line. We found Emax at RR1/RR2 = 1 on the regression line to be virtually identical with both mean Emax during AF and stable Emax obtained during irregular atrial pacing at the same intervals as the mean RR interval during AF. These results newly indicate that the pressure-interval relationship predominantly characterizes LV irregular beat contractilities and their mean level during AF.

beta-Adrenergic receptor (betaAR) downregulation and desensitization are hallmarks of the failing heart. However, whether abnormalities in betaAR function are mechanistically linked to the cause of heart failure is not known. We hypothesized that downregulation of cardiac betaARs can be prevented through inhibition of PI3K activity within the receptor complex, because PI3K is necessary for betaAR internalization. Here we show that in genetically modified mice, disrupting the recruitment of PI3K to agonist-activated betaARs in vivo prevents receptor downregulation in response to chronic catecholamine administration and ameliorates the development of heart failure with pressure overload. Disruption of PI3K/betaAR colocalization is required to preserve betaAR signaling, since deletion of a single PI3K isoform (PI3Kgamma knockout) is insufficient to prevent the recruitment of other PI3K isoforms and subsequent betaAR downregulation with catecholamine stress. These data demonstrate a specific role for receptor-localized PI3K in the regulation of betaAR turnover and show that abnormalities in betaAR function are associated with the development of heart failure. Thus, a strategy that blocks the membrane translocation of PI3K and leads to the inhibition of betaAR-localized PI3K activity represents a novel therapeutic approach to restore normal betaAR signaling and preserve cardiac function in the pressure overloaded failing heart.

. Jeffrey J. Nienaber and Hideo Tachibana contributed equally to this work. Conflict of interest:The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: β-adrenergic receptor (βAR); G protein-coupled receptors (GPCRs); phosphatidylinositol (PtdIns); phosphoinositide kinase domain (PIK); phosphatidylinositol-3,4,5-tri-phosphate (PtdIns-3,4,5-P3); inactive mutant of PI3Kγ (PI3Kγinact); PI3Kγ knockout (PI3Kγ-KO); hemagglutinin (HA); transverse aortic constriction (TAC); left ventricle (LV); PtdIns-3,4,5-tri-phosphate (PIP3); phosphoprotein kinase B (pPKB); phospho-glycogen synthase kinase (pGSK); extracellular signal-regulated kinase (ERK); LV weight/body weight (LVW/BW); isoproterenol (ISO); immunoblotting (IB); c-terminal region of β-adrenergic receptor kinase-1 (βARK1-ct).

Recently we have shown that the left ventricular end-systolic pressure-volume relation (ESPVR) of in situ rat hearts is an upward convex curve in contrast to the linear left ventricular ESPVR in dog and human hearts. Within the smaller left ventricular volume range, the left ventricular end-systolic pressure rose steeply with increases in left ventricular volume, but it gradually reached a plateau at the larger left ventricular volumes. In adult rat hearts, the myosin isozyme is V1, unlike V3 in dog and human hearts. To investigate whether myosin isozyme affects the curvilinearity of the left ventricular ESPVR, we evaluated the left ventricular ESPVR in hypothyroid rats in which the left ventricular myosin isozyme had been shifted to V3. In the hypothyroid rats, the left ventricular contractility was depressed and the ESPVR became closer to linear. However, after dobutamine administration the ESPVR returned to curvilinear. In nor-mal rats the curvilinearity of the left ventricular ESPVR was decreased by negative inotropic agents such as adrenergic blockers. These results indicate that the depressed left ventricular contractility in the hypothyroidism make ESPVR linear and that the enhanced left ventricular contractility from dobutamine make it curvilinear. We concluded that the curvilinearity of the rat left ventricular ESPVR is not determined by myosin isozyme per se, but by the left ventricular contractility.