bluuebunny/crossref_metadata_2025_split huggingface dataset:
Canagliflozin Inhibits Human Endothelial Cell Inflammation via the Induction of Heme Oxygenase-1
Authors: PEYTON KELLY J. , BEHNAMMANESH GHAZALEH , DURANTE WILLIAM 
Published: June 2018

Vascular disease is the major cause of mortality in diabetes. Although many factors contribute to diabetes-associated vascular disease, inflammation of endothelial cells (ECs) plays a critical role. Sodium-glucose transporter 2 (SGLT2) inhibitors are recently approved anti-hyperglycemic drugs that cause a significant reduction in cardiovascular events in type 2 diabetic patients, but the mechanisms underlying this protection are not fully known. In the present study, we investigated the effect of SGLT2 inhibitors on the expression of the vasoprotective protein heme oxygenase-1 (HO-1) in human umbilical vein ECs. Treatment of ECs with canagliflozin, dapagliflozin, or empagliflozin stimulated the expression of HO-1 mRNA and protein; however, only clinically achievable concentrations of canagliflozin (3-10µM) increased HO-1 levels. The induction of HO-1 by canagliflozin was associated with an increase in Nrf2 activity and reactive oxygen species (ROS) formation. Canagliflozin also stimulated HO-1 promoter activity, and this was prevented by mutating the antioxidant responsive element or by overexpressing dominant-negative Nrf2. In addition, the canagliflozin-mediated induction of HO-1 was abolished by N-acetyl-L-cysteine or rotenone. Significantly, prior treatment of ECs with canagliflozin inhibited the induction of the adhesion molecules ICAM-1 and VCAM-1 in response to high concentrations of glucose (25mM) and oxidized low-density lipoprotein (50mg/L) that mimic the diabetic milieu. Significantly, the anti-inflammatory action of canagliflozin was abrogated by blocking HO-1 activity or expression.
In conclusion, this study demonstrates that canagliflozin stimulates HO-1 gene expression in the vascular endothelium via the ROS-Nrf2 pathway, and that canagliflozin mitigates inflammation in human ECs exposed to a diabetic environment via the induction of HO-1. Thus, canagliflozin may exert some of it cardiovascular benefit in diabetes by targeting HO-1.

Disclosure
K.J. Peyton: None. G. Behnammanesh: None. W. Durante: None.
Actions of Liraglutide on Vascular Endothelial Cells Play a Central Role in the Suppression of Atherosclerosis in Diabetic Mice
Authors: HIROMURA MUNENORI , MORI YUSAKU , KOSHIBU MASAKAZU , KUSHIMA HIDEKI , KOHASHI KYOKO , TERASAKI MICHISHIGE , HIRANO TSUTOMU 
Published: June 2018

Background: Glucagon-like peptide-1 receptor agonists (GLP-1RAs) have been shown to exert direct anti-atherosclerotic effects through multiple actions on various cells. However, it is unclear which action is crucial. We investigated the role of the anti-atherogenic effects of liraglutide (Lira) on vascular endothelial cells (VECs) in vivo.
Methods: Streptozotocin-induced diabetic apolipoprotein E-null mice (male, 20 weeks) were randomly assigned to treatment with saline or Lira (17 nmol/kg/d) and underwent femoral artery (FA) wire injury to remove VECs. Thioglycolate-exudated peritoneal macrophages and vascular samples (aorta, FA, and right brachiocephalic artery [RBCA]) were collected 4 weeks after treatment.
Results: In the Lira group, active GLP-1 levels were significantly higher (4.7 ± 1.1 vs. 2.8 ± 0.3 pmol/L) and levels of HbA1c (8.9 ± 0.4% vs. 8.3 ± 0.5%) and total cholesterol (492 ± 8 vs. 457 ± 19 mg/dL) were significantly lower than in controls. Lira significantly reduced the atherosclerotic plaque (oil red O [ORO]-stained area) on the surface of the whole aorta by 36% and the plaque volume and intraplaque macrophage infiltration in the aortic root by 56% and 55%, respectively. Additionally, Lira suppressed the expression of inflammatory cytokines in macrophages by 40-45% (p &lt; 0.05) and the expression of inflammatory cytokines in the RBCAs, which are prone to lesion, plaque formation, and rupture. In FA, no plaque was observed in uninjured lesions, while severe plaque formation occurred in injured lesions. In contrast, Lira failed to suppress plaque volume or neointimal thickening in FAs where the VECs were removed (ORO-stained area/vascular wall area: 0.51 ± 0.vs. 0.55 ± 0.08, neointimal area/medial area: 2.4 ± 1.2 vs. 2.6 ± 0.7), indicating that Lira cannot suppress atherosclerosis independent of VECs.
Conclusions: Of the multiple actions on various cells, VECs play a central role in the suppression of atherosclerosis mediated by Lira.

Disclosure
M. Hiromura: None. Y. Mori: None. M. Koshibu: None. H. Kushima: None. K. Kohashi: None. M. Terasaki: None. T. Hirano: Speaker's Bureau; Self; Novo Nordisk Inc., AstraZeneca.
Coronary Microvascular Smooth Muscle Cells Contribute to Heart Failure in Diabetes Mellitus by Increasing Methylglyoxal Flux
Authors: BIDASEE KESHORE 
Published: June 2018

Prognosis for patients with type 1 diabetes mellitus (T1DM) who have heart failure is poor because of an incomplete understanding of molecular cues that trigger it and a lack of specific pharmacologic agents/strategies to slow/reverse its development. Supra-physiologic levels of the α-oxoaldehyde methylglyoxal (MG) have emerged as a possible cue. However, its source remains poorly characterized. Here we investigated MG levels and expression of its degrading and synthesizing enzymes glyoxalase-I (Glo-I) and vascular adhesion protein-1 (VAP-1) in ventricular tissues from rats with chronic streptozotocin-induced T1DM. We also used a gene transfer approach to prevent Glo-I downregulation in ventricular tissues to determine if it would be cardio-protective. When compared to controls, hearts of rats with chronic T1DM (10 -12 weeks) had 5-fold higher MG in ventricular tissues, 2.2-fold lower level of Glo-I in ventricular tissues and 2.5-fold higher in coronary microvascular smooth muscle cells (cSMCs). Preventing Glo-I levels downregulation in myocytes and cSMCs of rats by injecting them with an adeno-associated virus contain Glo-I driven by the endothelin promoter one week after the onset of T1DM, attenuated ventricular MG increase, with no effect in blood MG. Increasing Glo-I also blunted VAP-1 upregulation in cSMCs, attenuated microvascular leakage, inflammation, and fibrosis. These data support the notion that an increase in MG arising in part from increased expression of VAP-1 is contributing to HF development in T1DM rats and lowering ventricular MG levels T1DM rats is cardio-protective.

Disclosure
K. Bidasee: None.
Rho-Kinase Induces CTGF Expression through Actin Dynamics in Mesangial Cells
Authors: NAGAI YOSUKE , KAWANAMI DAIJI , MATOBA KEIICHIRO , TAKEDA YUSUKE , AKAMINE TOMOYO , ISHIZAWA SHO , KANAZAWA YASUSHI , YOKOTA TAMOTSU , UTSUNOMIYA KAZUNORI 
Published: June 2018

Introduction: Transforming growth factor (TGF)-β promotes the development of diabetic nephropathy by enhancing excessive production of extracellular matrix such as connective tissue growth factor (CTGF) in mesangial cells. However, the mechanism by which TGF-β induces CTGF expression has not been elucidated. In this study, we aimed to investigate contribution of small GTPase Rho and Rho-kinase to TGF-β-mediated CTGF expression in mesangial cells.
Methods and Results: Exposure of mouse mesangial cells (MES-13) to TGF-β induced activation of Rho and Rho-kinase. Y-27632, a specific Rho-kinase inhibitor, reduced TGF-β-mediated CTGF protein expression, indicating that TGF-β induces CTGF expression via Rho-kinase activation. Western blot analysis demonstrated that Y-27632 attenuates TGF-β-mediated phosphorylation of Erk1/2 and JNK/SAPK and nuclear translocation of NF-κB. Rho-kinase has been shown to regulate cytoskeleton reorganization. We therefore investigated whether Rho-kinase is involved in actin dynamics-induced NF-κB activation. Immunostaining studies demonstrated that Y-27632 and Cytochalasin D, an inhibitor of actin polymerization, attenuated TGF-β-mediated nuclear translocation of NF-κB, indicating that Rho-kinase-mediated actin reorganization regulates NF-κB activation. Furthermore, siRNA-mediated knockdown of Rho-kinase isoforms demonstrated ROCK2 but not ROCK1 is involved in these observations. In the present study, we demonstrate for the first time that Rho-kinase, especially ROCK2, mediates TGF-β-induced expression of CTGF via phosphorylation of Erk1/2 and JNK/SAPK and subsequent actin reorganization-mediated nuclear translocation of NF-κB in mesangial cells.
Conclusion: Rho-kinase could be an important therapeutic target against diabetic nephropathy.

Disclosure
Y. Nagai: None. D. Kawanami: None. K. Matoba: None. Y. Takeda: None. T. Akamine: None. S. Ishizawa: None. Y. Kanazawa: None. T. Yokota: None. K. Utsunomiya: None.
Dietary Intervention for Inpatient Glycemic Control
Authors: CROMER SARA , BOGUN MAGDALENA M. 
Published: June 2018

While recommendations for inpatient glycemic control suggest a conservative target of 140-180mg/dL, many struggle to achieve this goal. Glycemic control begins with a carbohydrate-controlled diet, but many inpatient diets continue to be suboptimal and can contribute to hyperglycemia. At our institution, the carbohydrate-controlled breakfast diet was changed to better reflect carbohydrate goals (e.g., replacing french toast on the “bad diet” with vegetable frittatas on the “good diet”). Pre- and post-prandial glucose values for patients with types I and II diabetes receiving a carbohydrate-controlled diet were collected by retrospective chart review during the weeks before and after these changes took effect. Patients on steroids were analyzed separately. Individual post-prandial glucose values were recorded in a binary fashion as meeting or exceeding predefined glucose targets (&lt;180 mg/dL, &lt;250mg/dL, and &lt;350mg/dL). During this period, 369 patients not receiving steroids and 39 patients receiving steroids had at least one relevant glucose value. In the non-steroid group, there was no difference in proportion of patients who exceeded any of the predefined thresholds. However, in the steroid group, there was a trend toward reduction in proportion of patients exceeding blood glucose of 180mg/dL (absolute risk reduction 19%, 95% confidence interval -4% to 42%, p = 0.12) and 250mg/dL (absolute risk reduction 17%, 95% confidence interval -2% to 35%, p = 0.08) on the days after the dietary change. Although all patients with diabetes may experience hyperglycemia proportionate to their carbohydrate intake, those on steroids experience induced insulin resistance leading to especially diet-sensitive glucose excursions. Our data suggests that patients receiving steroids can likely achieve improved inpatient glucose control through dietary modification. Physician and administration attention to carbohydrate-controlled menus represents an opportunity for improved quality of care for inpatients with diabetes.

Disclosure
S. Cromer: None. M.M. Bogun: Other Relationship; Self; Mytonomy.
DNA Hydroxymethylation Regulates Beta-Cell Maturation and Expansion
Authors: GURLO TATYANA , GEORGIA SENTA , DHAWAN SANGEETA 
Published: June 2018

Epigenetic mechanisms play a central role in governing functional beta-cell mass. In particular, the methylation of DNA to generate 5-methylcytosine (5mC), directed by DNA methyltransferases, has been shown to regulate the establishment and maintenance of beta-cell identity and function. While the contribution of DNA methylation to beta-cell homeostasis and diabetes pathogenesis has been studied in detail, not much is known about the role of DNA demethylation in beta-cells. Oxidative conversion of 5mC to 5-hydroxymethylcytosine (5hmC) by Ten Eleven Translocation (Tet) enzymes constitutes a key step towards active DNA demethylation. In the present study, we provide evidence that the 5hmC landscape is dynamic, and undergoes specific changes during embryonic and postnatal islet development, islet adaptation, and in diabetes. We report that the Pdx1-positive pancreatic progenitors are marked by very low 5hmC, and beta-cell differentiation and functional maturation is accompanied by a progressive increase in 5hmC. However, under conditions during adaptive beta-cell expansion as well as in diabetes 5hmC levels decline in part due to the oxidative stress. These changes in 5hmC pattern are conserved in mouse and human pancreas, and are accompanied by the changes in Tet2 levels. We show that deletion of Tet2 in the pancreatic lineage results in improved glucose tolerance and beta-cell function. Together, our data show the importance of Tet2-dependent 5hmC patterning in maintaining a differentiated beta-cell phenotype, and highlight the remodeling of beta-cell epigenetic landscape as a key component of diabetes pathogenesis.

Disclosure
T. Gurlo: None. S. Georgia: None. S. Dhawan: None.
Renin Production and Involvement of ß1 Integrin in Blood Pressure Regulation in Diabetic Nephropathy
Authors: TOYODA MASAO , SAITO NOBUMICHI , KIMURA MORITSUGU , MORIYA HITOMI , SAWADA KAICHIRO , FUKAGAWA MASAFUMI 
Published: June 2018

Diabetic nephropathy (DN) and hypertension mutually exacerbate each other, but many aspects of the mechanisms of these two pathologies remain unclear. The juxtaglomerular cells (JGC) secrete renin and trigger the renin-angiotensin-aldosterone system involved in blood pressure regulation. Integrins (Itgs) are cell surface proteins that interact with the extracellular matrix and transduce intracellular signals in response to extracellular stimuli. There are several phosphorylation sites in the β chain, and phosphorylation plays an important role in the cell-binding ability and intracellular signaling functions of Itgs. Our previous immunohistochemical analysis of renal tissue sections of diabetic patients showed that β1 Itg, a cellular matrix adhesion factor, undergoes JGC-specific phosphorylation. To characterize the relationship between the phosphorylated β1 (pβ1) Itg and renin expression in DN, the following studies were conducted in rats with streptozotocin (STZ)-induced diabetes.
Immunohistochemical analysis of the prepared sections showed JGC-specific phosphorylation of β1 Itg in the kidneys of diabetic rats. Immunofluorescent staining of these kidney tissue specimens confirmed the coexpression of pβ1 Itg and renin in JGC.
The mRNA levels of pβ1 Itg decreased continuously with time in diabetic rats after the onset of diabetes, till month 2. However, the mRNA levels of pβ1 Itg increased subsequently, but ultimately showed recovery to the baseline level. In contrast, the renin mRNA ratio exhibited an inverse correlation with pβ1 Itg. Quantification by image analysis of confocal laser microscopic images confirmed the negative correlation between pβ1 Itg and renin.
Our results showed a negative correlation between pβ1 Itg and renin expression in renal JGC, suggesting a link between β1 Itg and the regulatory system that controls renin production and secretion.

Disclosure
M. Toyoda: None. N. Saito: None. M. Kimura: None. H. Moriya: None. K. Sawada: None. M. Fukagawa: None.
Klotho Prevents Epithelial-Mesenchymal Transition through Egr-1 Down-Regulation in Diabetic Kidney Disease
Authors: LI YANG , XUE MENG , HU FANG , JIA YIJIE , YANG YAN-LIN , XUE YAO-MING 
Published: June 2018

Background: As a key event leading to tubulointerstitial fibrosis in diabetic kidney disease (DKD), epithelial-mesenchymal transition (EMT) has drawn increasing attention from researchers. Klotho attenuates renal fibrosis in part by inhibiting ERK1/2 signaling in DKD. Early growth response factor 1 (Egr-1), activated mainly by ERK1/2, has been shown to play an important role in EMT. However, whether Klotho prevents EMT by inhibiting ERK1/2-dependent Egr-1 expression in DKD is unclear. The aim of this study was to investigate whether Klotho prevents EMT through Egr-1 down-regulation by inhibiting the ERK1/2 signaling pathway in DKD.
Methods: Male C57BL/6J mice fed an HFD for 4 weeks received 120 mg/kg STZ. Klotho and Egr-1 expression was detected in the renal cortices of these mice upon their sacrifice at 6 and 12 w after STZ treatment. In vitro studies, we incubated HK2 cells under high-glucose (HG) or TGF-β1 conditions to mimic DKD. We then transfected the cells with an Egr-1-containing plasmid, Egr-1 siRNA, a Klotho-containing plasmid or Klotho siRNA. Klotho, Egr-1, E-cadherin, α-smooth muscle actin (α-SMA), fibronectin(FN), ERK1/2 and p-ERK1/2 were determined.
Results: Klotho expression was significantly decreased in the renal cortices of diabetes mellitus (DM) mice compared to control mice at 6 w and was even more significantly decreased at 12 w. In contrast, Egr-1 expression was significantly increased in DM mice compared to control mice only at 12 w. We also found that Klotho overexpression down-regulated Egr-1 expression and the (p-ERK1/2)/(ERK1/2) ratio in HG- or TGFβ1-treated HK2 cells. Conversely, Klotho silencing up-regulated Egr-1 expression and the (p-ERK1/2)/(ERK1/2) ratio. Moreover, the effects of si-Klotho were abolished by the ERK1/2 inhibitor PD98059.
Conclusions: Klotho prevents EMT during DKD progression, an effect that has been partially attributed to Egr-1 down-regulation mediated by ERK1/2 signaling pathway inhibition.

Disclosure
Y. Li: None. M. Xue: None. F. Hu: None. Y. Jia: None. Y. Yang: None. Y. Xue: None.
Serpin B13 Plays a Role in Beta-Cell Development and Progression to Insulin-Dependent Diabetes
Authors: KRYVALAP YURY , JIANG MATTHEW L. , KRYVALAP NADZEYA , MUELLER KYLE A. , CZYZYK JAN 
Published: June 2018

Identifying natural factors of the progression to type 1 diabetes (T1D) provides a unique opportunity to develop novel preventative strategies for this disease. Our laboratory has identified serpin B13 proteinase inhibitor as a potential player, activity of which is involved in modifying the timing of T1D development. In this study we examined whether blocking of serpin B13 with a monoclonal antibody (mAb) in early life causes any changes in pancreatic islet development that may explain beneficial impact of this activity later in life. First, our animal studies showed that blocking of serpin B13 during mid-pregnancy caused robust increase in the number of Ngn3+ progenitor cells at day E16.5 and accelerated conversion of these cells to beta cells. Similar changes were observed in embryonic pancreatic buds in vitro. On the other hand, the serpin B13 protein significantly downregulated the output of Ngn3+ cells in these in vitro cultures. Importantly, at 2 to 3 months after birth, the offspring of serpin B13 mAb-treated mothers demonstrated increased islet cell mass and responded to the diabetes-inducing agent, streptozotocin, with markedly improved glucose tolerance and renal function, compared with control animals. Second, our screening of 280 children, previously enrolled in a DPT-1 clinical trial, for serpin B13 autoantibody (AA) revealed an inverse correlation of this AA with risk level for T1D, as well as a positive association with longer diabetes-free interval. These data demonstrate the significant, long-term impact of serpin B13 activity on islet biology and suggest that blocking this serpin has the potential to partially prevent, or at least slow down, the development of T1D both in the mouse and human.

Disclosure
Y. Kryvalap: None. M.L. Jiang: None. N. Kryvalap: None. K.A. Mueller: None. J. Czyzyk: None.
Klotho Prevented Epithelial-to-Mesenchymal Transition in High-Fat Diet/Streptozotocin-Induced Mice by Targeting NFkB/Microrna-21/Smad7 Signaling Pathway
Authors: XUE MENG , HU FANG , LI YANG , JIA YIJIE , XUE YAO-MING 
Published: June 2018

Klotho is an antiaging hormone present in the kidney that protect renal via attenuating the nuclear factor-kB (NFkB) p65 activity. And in kidney epithelium miRNAs become active in response to injury. But little is known regarding to the role of Klotho on miRNAs in the pathogenesis of diabetic nephropathy(DN). In this study, we found that the expression of Klotho was reduced with upregulation of miR-21 and key parameters of DN in HFD/STZ-induced diabetic mice. Klotho-treatment reduced p65 nucleus expression in association with modulation of miR-21/Smad7 and amelioration of fibrosis. Furthermore, we found that Klotho expression was decreased in high glucose induced HK2 at both mRNA and protein, accompanied by the significantly increased miR-21 expression. NF-κB was demonstrated to regulate miR-21 expression by directly binding to its promotor. Additionally, exogenous addition of Klotho or inhibition of NF-kB restrained the activity of miR-21/Smad7 signaling pathways, which suppressed fibrosis in high glucose cultured HK2. This study provides a new basis to elucidate the protection mechanism of anti-aging protein Klotho in diabetic kidney. We firstly found that Klotho treatment could delay the fobrosis progression of diabetic nephropathy in type 2 diabetic mice via inhibition of NF-kB/miR-21/Smad7 signaling cascade pathways.

Disclosure
M. Xue: None. F. Hu: None. Y. Li: None. Y. Jia: None. Y. Xue: None.
Empagliflozin and Linagliptin Alleviate Glomerular Changes in a Model of Diabetic Nephropathy
Authors: KORBUT ANTON I. , KLIMONTOV VADIM , TASKAEVA IULIIA , BGATOVA NATALIYA P. , ISHCHENKO IRINA , ZAVJALOV EVGENII L. 
Published: June 2018

The aim: to assess the effects of SGLT2 inhibitor empagliflozin, DPP-4 inhibitor linagliptin, and their combination on structural changes in kidneys in a model of type 2 diabetic nephropathy. Eight-week-old male db/db diabetic mice (BKS.Cg-Dock7m+/+Leprdb/J) were treated with empagliflozine (10 mg/kg), linagliptin (10 mg/kg), combination of both agents, or placebo for 8 weeks. Nondiabetic heterozygous db/+ mice were used as control. Renal structural changes were analyzed quantitatively from the light and electron microscopic images. Both empagliflozin and linagliptin, either alone or in combination, attenuated mesangial expansion estimated by mesangial volume (p=0.00for empagliflozin, p=0.03 for linagliptin and combination). The width of glomerular basement membrane was diminished in all treated groups (all p&lt;0.01). In diabetic mice, podocytopathy was manifested by the foot process effacement (p=0.03). Under treatment, the number of podocyte foot processes increased (p=0.003 for linagliptin, p=0.03 for empagliflozin and combination) and the width of podocyte foot processes decreased in all treated groups (p=0.003 for empagliflozin and linagliptin, p=0.001 for combintion). The number of endothelial fenestrae in glomerular capillaries was increased after the treatment (all p&lt;0.01). The interaction between empagliflozin and linagliptin was significant for the number and width of podocyte foot processes, and the number of endothelial fenestrae (all p&lt;0.01). The effects of both agents on structural changes were not associated with hypoglycemic activity. The obtained results demonstrate that empagliflozin and linagliptin ameliorate renal fibrosis and podocyte injury in a model of type 2 diabetic nephropathy.

Disclosure
A.I. Korbut: None. V. Klimontov: None. I. Taskaeva: None. N.P. Bgatova: None. I. Ishchenko: None. E.L. Zavjalov: None.