bluuebunny/crossref_metadata_2025 huggingface dataset:
Single and Combined Effects of Peripheral Artery Disease and of Type 2 Diabetes Mellitus on the Risk of Cardiovascular Events in Women
Authors: SAELY CHRISTOPH H. , SCHINDEWOLF MARC , VONBANK ALEXANDER , HEINZLE CHRISTINE , ZANOLIN DANIELA , LARCHER BARBARA , MADER ARTHUR , LEIHERER ANDREAS , MUENDLEIN AXEL , DREXEL HEINZ , BAUMGARTNER IRIS
Published: June 2018

Both peripheral artery disease (PAD) and type 2 diabetes (T2DM) are associated with a high risk of cardiovascular events. However, the single and combined effects of PAD and of T2DM in women have not been investigated yet and are addressed in the present study. We prospectively recorded cardiovascular events in a series of 436 women of whom 94 had PAD and 342 did not have PAD. At baseline, the prevalence of diabetes was higher in women with PAD than in those who did have PAD (41.5 vs. 21.6%, p<0.001). Over a mean follow-up period of 7.2 years we recorded 1cardiovascular events. When compared to the event rate in women with neither PAD nor T2DM (14.9%) the event rate was not significantly increased in those with T2DM but without PAD (16.2%; p=0.972) but was significantly higher in nondiabetic women with PAD (47.3%; p<0.001) and further increased in those with both PAD and T2DM (76.9%; p<0.001). Nondiabetic women with PAD were at a significantly higher cardiovascular risk than women with T2DM who did not have PAD (p=0.001). When compared to women with neither PAD nor T2DM, adjusted hazard ratios were 0.93 [0.48-1.80]; p=0.831, 3.80 [2.02-7.13]; p<0.001, and 7.41 [3.87-14.19]; p<0.001 for women with T2DM only, for those with PAD only and for those with the combination of PAD plus T2DM, respectively. We conclude that T2DM strongly increases the risk of future cardiovascular events in women with PAD. However, type 2 diabetic women who do not have PAD are at a significantly lower cardiovascular event risk than nondiabetic women with PAD. PAD in women is a stronger risk factor than T2DM.

Disclosure

C.H. Saely: None. M. Schindewolf: None. A. Vonbank: None. C. Heinzle: None. D. Zanolin: None. B. Larcher: None. A. Mader: None. A. Leiherer: None. A. Muendlein: None. H. Drexel: None. I. Baumgartner: None.
NREP Bridges TGF-ß Signaling and Lipid Metabolism in the Epigenetic Reprogramming of NAFLD in the Offspring of Insulin-Resistant Parents
Authors: JESUS DARIO F. DE , ORIME KAZUKI , WANG CHIH-HAO , HU JIANG , DIRICE ERCUMENT , SILVA AMÉLIA M. , TSENG YU-HUA , PIHLAJAMAKI JUSSI , KULKARNI ROHIT
Published: June 2018

Nonalcoholic fatty liver disease (NAFLD) prevalence is increasing worldwide and few studies have associated maternal diabetes and birth weights with increased risk for NAFLD. We used an unique non-dietary model, manifesting hyperglycemia and hyperinsulinemia-two hallmarks of gestational and type 2 diabetes. We aimed to determine the genetic and epigenetic effects of paternal vs. maternal genetic insulin resistance on the developmental programming in the offspring of the liver-specific insulin receptor knockout (LIRKO) mice. Male control F1 offspring from father LIRKO (FL), mother LIRKO (ML) or control mothers and fathers (C) were weaned on a chow or high-fat-diet (HFD) and followed for 3 months. FL and ML showed impaired growth and body weight composition. FL and ML developed hepatic steatosis compared to C when challenged with HFD and exhibited increased hepatic expression of lipogenesis-associated genes. Hepatic transcriptomic and genome-wide DNA methylation analyses of FL and ML on chow diet presented enriched-pathways associated with TGF-β signaling and lipid synthesis. FL and ML hepatic NREP mRNA levels were decreased 50% (p<0.05) on HFD compared to C. In-vivo and in-vitro modeling of hepatic steatosis in HepG2 and human primary hepatocytes decreased NREP mRNA and protein levels. Knock-down experiments performed in HepG2 cells revealed that NREP acts by regulating triglyceride and cholesterol synthesis transcriptional network via regulation of ATP-citrate lyase (ACL) in a phospho-AKT dependent manner. Finally, reduced NREP mRNA levels in patients with hepatic steatosis by 40% (p<0.05) and recent preclinical trials implicating ACL in NAFLD highlight the translation relevance of our findings. These data suggest that prenatal insulin resistance epigenetically regulates a novel gene, NREP, that has detrimental effects on metabolic adaptation and transcriptional regulation of hepatic metabolism in the development of NAFLD.

Disclosure

D.F. De Jesus: None. K. Orime: None. C. Wang: None. J. Hu: None. E. Dirice: None. A.M. Silva: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd.. Research Support; Self; MedImmune. J. Pihlajamaki: None. R. Kulkarni: None.
Bone and Vascular Health in Postmenopausal Women with Type 1 Diabetes
Authors: SHAH VIRAL N. , JOSHEE PRAKRITI , SIPPL RACHEL M. , SNELL-BERGEON JANET K.
Published: June 2018

The risk for cardiovascular (CV) disease and osteoporotic fractures is higher in postmenopausal women with type 1 diabetes (T1D). However, it is unclear whether these processes have common risk factors. We examined the associations between bone mineral density (BMD), trabecular bone score (TBS), and CV risk measures in 47 postmenopausal women with T1D (62 years, diabetes duration; 41 years and A1c 7.8%) and 51 postmenopausal women without diabetes (64 years and A1c 5.5%).

Age, menopausal duration, BMI, serum creatinine, calcium, 25-OH-D, parathyroid hormone (PTH) and bone turnover markers (BTMs) were similar between two groups. T1D postmenopausal women had lower left femoral neck and distal forearm BMD, and subendocardial viability index (SVI) and higher augmented pressure (AP) and carotid intima media thickness (cIMT) compared to controls (Table).

HbA1c was negatively correlated with distal forearm BMD, TBS and SVI and positively correlated with AP. Total cholesterol and LDL-C were associated with arterial stiffness but not with BMD. PTH and BTMs were associated with lower BMD but not with CV risk measures. Urinary microalbumin was inversely correlated with distal forearm BMD and TBS, and positively correlated with AP and cIMT.

T1D is associated with lower femoral neck and distal forearm BMD and higher arterial stiffness and cIMT. Urine microalbumin was the only factor linked to both low BMD and high CV risk.VariablesPostmenopausal women with T1D (n=47)Postmenopausal women without diabetes (n=51)P-valueLeft femoral neck BMD (g/cm2)0.6±0.10.7±0.10.01Left distal forearm BMD (g/cm2)0.61±0.090.66±0.080.008Lumbar spine BMD(g/cm2)0.9±0.10.9±0.10.9Subendocardial viability index (%)119.0±18.1149.1±23.9<0.001Augmented pressure (mmHG)16.4±6.013.3±5.10.006Carotid intima media thickness (mm)0.7±0.10.6±0.090.01

Disclosure

V.N. Shah: None. P. Joshee: None. R.M. Sippl: None. J.K. Snell-Bergeon: Stock/Shareholder; Self; Abbott. Research Support; Self; Roche Diagnostics Corporation.
A Comparative Study of the Associations between Modified Low-Density Lipoproteins and Vascular Function in African-American Diabetic Patients
Authors: ADEDAYO AJIBOLA M. , ELUWOLE AYOBAMI , TEDLA FASIKA , KREMER ARYE , MASTROGIOVANNI NICOLE , ROSENBERG CARL , DREIZEN PAUL , LAROSA JOHN , SALCICCIOLI LOUIS , BOUTJDIR MOHAMED , BANERJI MARY ANN , BROWN CLINTON , SALIFU MORO , LAZAR JASON , BAKILLAH AHMED
Published: June 2018

Background: LDL plays a key role in development and progression of atherosclerosis. Elevated LDL amplifies the atherosclerotic process but the significance of modified LDL in pathogenesis of vascular complications is unclear. We aimed to assess plasma modified LDL levels and their relationship with vascular function in African American diabetic patients.

Methods: 125 patients with type-2 diabetes were enrolled. Levels of glycated LDL (g-LDL), carbamylated LDL (c-LDL), nitrated LDL (n-LDL) and oxidized LDL (ox-LDL) were quantified by ELISA. Microvascular function was assessed by vascular reactivity index (VRI), which assesses changes in digital temperature before and after release of arterial cuff occlusion. Carotid-femoral pulse wave velocity (PWV) assessed arterial stiffness using applanation tonometry. B-mode ultrasound image analysis assessed Carotid intima-media thickness (CIMT). Patient population was divided into well-controlled: HbA1c ≤7.0%, N=54; poorly-controlled: HbA1c >7.0%, N=71.

Results: Age 60±8 years; 64% Female, 80% Hypertension, 90% Dyslipidemia and 15% CKD. HbA1c level 8.1±2.2%, diabetes duration 10.29±3.79 years. Mean plasma g-LDL, c-LDL, n-LDL and ox-LDL were 1.56±0.64 mg/dL, 0.85±0.11 mg/dL, 2.33±0.39 ng/mL, and 61.93±9.25 ng/mL. Ox-LDL was positively correlated with CIMT in well-controlled patients (r= 0.45, p=0.001), but not in poorly-controlled (r= 0.12, p=0.30). Multi-regression analysis revealed ox-LDL was independently associated with CIMT, but neither with PWV nor VRI after adjustment for variables such as age, gender, weight, smoking, total cholesterol, HDLc, triglycerides, and LDLc (β= 0.003, p= 0.012; r2= 0.38).

Conclusion: Ox-LDL offered better predictive value for CIMT in well-controlled patients than other forms of modified LDL. These data favor ox-LDL as potential marker to identify diabetic patients at risk of developing atherosclerotic vascular complications.

Disclosure

A.M. Adedayo: None. A. Eluwole: None. F. Tedla: None. A. Kremer: None. N. Mastrogiovanni: None. C. Rosenberg: None. P. Dreizen: None. J. LaRosa: None. L. Salciccioli: None. M. Boutjdir: None. M. Banerji: Speaker's Bureau; Self; Merck & Co., Inc.. C. Brown: None. M. Salifu: None. J. Lazar: None. A. Bakillah: None.
Protective Role of L-Cysteine Against High Glucose Induces G6PD-Deficiency and Endothelial Dysfunction
Authors: PARSANATHAN RAJESH , JAIN SUSHIL K.
Published: June 2018

African Americans have high incidences of diabetes and cardiovascular disease, and glucose-6-phosphate dehydrogenase (G6PD) deficiency. G6PD supplies NADPH, which recycles GSSG to GSH. Clinical studies demonstrate that lower blood GSH/G6PD correlate with excess oxidative stress with vascular inflammation markers which contribute to the development of complications in diabetic patients. However, the contribution of high glucose (HG) on G6PD deficiency to endothelial activation/dysfunction and monocyte recruitment, a forerunner of atherosclerosis, remains unknown. G6PD-normal and G6PD-deficient HUVEC and monocytic cells were silenced with control or G6PD siRNA (100nM) for 24 h, followed by exposure with normal glucose (5.5mM) or HG (25mM) or glutathione precursor L-Cysteine (LC; 300µM) in basal medium for 6 h. HG reduced the G6PD mRNA and its activity along with GSH in both G6PD-normal and G6PD-deficient endothelial and monocytic cells. Conversely, mRNA levels of iNOS, NOX4, and its activity were significantly upregulated but eNOS and nitrate levels were remarkably decreased under HG treated G6PD-deficient cells compared to G6PD-normal cells. The expression of cell adhesion molecules (CAMs) ICAM-1 and VCAM-1 mRNA levels significantly increased in HG-treated G6PD-deficient cells compared to G6PD-normal cells. Elevated MCP-1 was found in G6PD-deficient cells under HG conditions. Enhanced adherence of monocytes to HUVEC was observed in HG-treated G6PD-deficient cells (15.13%; p<0.005) compared to control cells (10%; ns). LC supplementation significantly increased the G6PD activity, GSH, and decreased CAMs reduced adherence to control level. These findings demonstrate that high glucose induces and fuels a GSH/G6PD-deficient phenotype to impair endothelial function; LC supplementation improves vascular reactivity by restoring GSH/G6PD levels.

Disclosure

R. Parsanathan: None. S.K. Jain: None.
Vasoprotective Effects of Vildagliptin in Mice Is Masked by Metformin—Overlapping Mechanisms Implied
Authors: KUSHIMA HIDEKI , MORI YUSAKU , KOSHIBU MASAKAZU , HIROMURA MUNENORI , KOHASHI KYOKO , TERASAKI MICHISHIGE , HIRANO TSUTOMU
Published: June 2018

Preclinical and small clinical studies have demonstrated the direct cardiovascular protective effects of dipeptidyl peptidase 4 inhibitors (DPP-4Is). However, cardiovascular benefits have not been proven in large clinical trials that utilized multiple antidiabetic agents. We hypothesized that the cardiovascular protective effects of the first-line antidiabetic agent metformin (Met) affects those of DPP-4Is, so we evaluated the vasoprotective effects of vildagliptin (Vil) in combination with Met in mice.

Methods: We randomly assigned C57BL6 and diabetic db/db mice (8 weeks old) to treatment with vehicle, Vil (3 mg/kg/d), Met (400 mg/kg/d), or the combination of Vil and Met. The mice were subjected to femoral artery wire injury to induce arterial remodeling. Four weeks after the injury, the femoral arteries were collected. Human umbilical vein endothelial cells (HUVECs) were used in the in vitro experiments.

Results: In wild type mice, the biological parameters were similar among the groups. Both Vil and Met treatment reduced neointimal area and vascular cell proliferation to a similar extent (40% reduction) compared to that of the vehicle, and combining the two drugs did not further reduce those values. Similarly, both Vil and Met treatment reduced neointimal area in db/db mice (50% reduction) without affecting biological parameters, but no additive effect was found. In wild type mice, cotreatment with the nitric oxide (NO) synthase inhibitor completely abolished the vascular effects of both Vil and Met, suggesting that Vil and Met had common mechanisms that are dependent on endothelial NO. In HUVECs, Vil did not directly affect NO production. In contrast, glucagon like peptide (GLP)-1 did stimulate NO production. Met also stimulated NO production. However, the combination of GLP-1 and Met did not increase NO production.

Conclusions: Metformin masks the vasoprotective effects of vildagliptin, possibly because of overlapping mechanisms that are dependent on endothelial NO.

Disclosure

H. Kushima: None. Y. Mori: None. M. Koshibu: None. M. Hiromura: None. K. Kohashi: None. M. Terasaki: None. T. Hirano: Speaker's Bureau; Self; Novo Nordisk Inc., AstraZeneca.
Inhibition of NF-κB Pathway with IKK-16 or Linagliptin Attenuates the Cardiac Dysfunction Associated with Polymicrobial Sepsis in Mice with Preexisting Type 2 Diabetes Mellitus (T2DM)
Authors: ZOUBI SURA AL , CHEN JIANMIN , MARTIN LUKAS , MURPHY CATHERINE , PURVIS GARETH S. , CHIAZZA FAUSTO , COLLOTTA DEBORA , COLLINO MASSIMO , THIEMERMANN CHRISTOPH
Published: June 2018

Introduction: Patients with diabetes are more susceptible to infections and sepsis. Activation of nuclear factor-κB (NF-κB) plays a substantial role in the pathophysiology of sepsis and diabetes. Here we investigate i) the effect of preexisting type 2 diabetes mellitus on cardiac dysfunction associated with sepsis and ii) whether inhibition of NF-κB using IKK-16 or linagliptin attenuates cardiac dysfunction in mice with sepsis and diabetes.

Methods: Ten-week old male C57BL/6 mice received high fat (HFD) or chow diet for 12 weeks, and were then subjected to caecal ligation and puncture (CLP) or sham surgery for 24 hours. At 1 hour after CLP, mice received IKK-16, linagliptin, or vehicle.

Results: Administration of HFD resulted in a significant impairment in glucose tolerance and a small, but significant, reduction in ejection fraction. Immunoblot analysis of hearts tissue showed that HFD caused significant NF-κB pathway activation and expression of inducible nitric oxide synthase (iNOS). Mice on HFD subjected to CLP showed further (i) decline in EF, (ii) increase in NF-κB pathway activation, (iii) increase in iNOS expression, (iv) an increase in serum ALT, and (v) a significant renal dysfunction. Treatment of HFD-CLP mice with IKK-16 or linagliptin resulted in significant reduction of the CLP-induced i) cardiac dysfunction, ii) NF-κB pathway activation, and iii) iNOS expression when compared to mice treated with vehicle.

Conclusion: Our results show that HFD results in inflammation and cardiac dysfunction. Moreover, a preexisting diabetic phenotype worsened the cardiac dysfunction associated with CLP-sepsis. Most notably, inhibition of NF-κB reduced the cardiac dysfunction caused by sepsis in animals with preexisting T2DM.

Disclosure

S. Al Zoubi: None. J. Chen: None. L. Martin: None. C. Murphy: None. G.S. Purvis: None. F. Chiazza: Other Relationship; Self; Boehringer Ingelheim GmbH. D. Collotta: None. M. Collino: None. C. Thiemermann: None.
The Effect of Resveratrol on Reducing Neointimal Growth after Femoral Artery Injury in AMPKα2 Knockout Mice Given High-Fat Diet
Authors: ZHOU LIWEI , GUO JUNE , ZHANG HANGJUN , HEXIMER SCOTT , GIACCA ADRIA
Published: June 2018

The rates of restenosis after percutaneous transluminal angioplasty are higher in diabetic than nondiabetic patients. Insulin is protective against restenosis, but these effects are diminished in the case of insulin resistance, which develops with high fat diet (HFD). Therefore, it makes sense to use an insulin-sensitizing compound which also has vasculoprotective effects such as resveratrol. Resveratrol is a phenolic compound found in plant tissues such as the skin of grapes, and has been hypothesized to account for the lower rates of cardiovascular disease among countries with high wine consumption. We have shown that resveratrol decreases neointimal formation in both mouse and rat models of arterial injury, and also that these effects are exerted through eNOS (endothelial nitric oxide synthase). We hypothesized that resveratrol acts though Sirt1, a NAD+-dependent deacetylase, to activate eNOS and decrease neointimal formation, but did not see any difference between neointimal growth of control and Sirt1-/- mice given HFD. AMPK (AMP-activated protein kinase) is an energy sensor which also protects against neointimal hyperplasia in the vasculature via directly activating eNOS and inhibiting the migration of vascular smooth muscle cells. Knocking out the α2 subunit of AMPK in particular has been shown to increase neointimal growth in mouse carotid artery in response to injury. We investigated the effect of resveratrol on neointimal growth after femoral arterial injury of wild type and AMPKα2 knock-out mice given HFD. Resveratrol was given at 0.0375% in the diet two days after wire injury. After 28 days, arteries were harvested and histomorphometric analysis was performed on the cross-sections. We found that neointimal growth was significantly higher in AMPKα2 knock-out mice given resveratrol compared to wild type mice given resveratrol (p<0.05). These results suggest that the vasculoprotective effects of resveratrol are mediated through AMPK.

Disclosure

L. Zhou: None. J. Guo: None. H. Zhang: None. S. Heximer: None. A. Giacca: None.
Tofogliflozin Suppresses the Progression of Atherosclerosis and Reduces Inflammation in Macrophages of STZ-Diabetic ApoE KO Mice due to Improvement of Glucose Tolerance
Authors: IWAMOTO MASAHIKO , KUBOTA NAOTO , KUBOTA TETSUYA , SAKURAI YOSHITAKA , WADA NOBUHIRO , TAKAMOTO ISEKI , SHIODA SEIJI , KADOWAKI TAKASHI
Published: June 2018

Background: Selective sodium-glucose cotransporter 2 inhibitor (SGLT2i) has become widely used for treatment of diabetes. Recently, EMPA-REG OUTCOME and CANVAS Program showed that SGLT2i reduce cardiovascular events in patients with type 2 diabetes, but the mechanism still remains uncertain. We investigated the mechanisms of vascular protection of Tofogliflozin (Tofo), one of the SGLT2i.

Method: 1) Streptozotocin (STZ)-diabetic apolipoprotein-E-deficient (ApoE KO) mice were separated into two groups, one group were fed normal chow and the other were fed normal chow containing 0.005% Tofo for ad libitum. After 6weeks, all mice were administered thioglycolate intraperitoneally, and 4 days later, peritoneal macrophages were collected to investigate the expressions of inflammatory cytokines of peritoneal macrophages by real-time PCR. Histological analyses were carried out by Sudan IV, Oil Red O, and MOMA-2 staining. 2) The same experiment targeted were carried out towards nondiabetic ApoE KO mice.

Results: 1) Tofo increased food and water intake and decreased blood glucose, but there were no significant changes in body weight, serum levels of free fatty acid, triglyceride and total cholesterol between two groups. The legions of atherosclerosis and macrophages accumulations were significantly suppressed in Tofo group. Moreover, the expression levels of TNF-α, IL-β were significantly suppressed in macrophages from Tofo group. 2) Nondiabetic ApoE KO mice showed similar increase in food and water intake, but failed to exhibit differences in blood glucose, Sudan IV staining, and expressions of inflammatory cytokines in macrophages, as observed in diabetic ApoE KO mice.

Conclusion: Tofogliflozin suppresses the progression of atherosclerosis and reduces inflammation in macrophages of STZ- diabetic ApoE KO mice. These beneficial effects may be exerted due to improvement of glucose tolerance.

Disclosure

M. Iwamoto: None. N. Kubota: None. T. Kubota: None. Y. Sakurai: None. N. Wada: None. I. Takamoto: None. S. Shioda: None. T. Kadowaki: Consultant; Self; Novo Nordisk A/S, AstraZeneca, Merck Sharp & Dohme Corp.. Research Support; Self; Kissei Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Sanofi, Kyowa Hakko Kirin Co., Ltd., Novo Nordisk A/S, Astellas Pharma, Daiichi Sankyo Company, Limited, Takeda, Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd., Merck Sharp & Dohme Corp., Nippon Boehringer Ingelheim Co. Ltd.. Speaker's Bureau; Self; Astellas Pharma, AstraZeneca, Merck Sharp & Dohme Corp., Ono Pharmaceutical Co., Ltd., Takeda, Eli Lilly and Company, Nippon Boehringer Ingelheim Co. Ltd., Novo Nordisk A/S.
Role of Hyaluronic Acid on Apolipoprotein A1 and HDL-Mediated Cholesterol Efflux in Macrophages
Authors: MATSUKI KOTA , OGURA MASATSUNE , HARADA-SHIBA MARIKO
Published: June 2018

Diabetic macroangiopathy are characterized by increased arterial stiffness and hyaluronic acid (HA) deposition in the large elastic arteries. It is noteworthy that systemic inhibition of HA synthesis by 4-methylumbelliferone interferes with the protective function of the endothelial glycocalyx, thereby facilitating leukocyte adhesion, inflammation, and progression of atherosclerosis. Apolipoprotein A1 (ApoA1) and High-density lipoprotein (HDL) mediates cholesterol efflux, which is the initial and rate-limiting step of reverse cholesterol transport preventing from atherosclerosis. The aim of the present study was to investigate the role of HA on ApoA1 and HDL-mediated cholesterol efflux in macrophages. Murine J774.1 macrophages and human THP-1 macrophages were labeled with 3H-cholesterol and incubated with ApoA1 (10 μg/ml) for 24 hours or HDL (50 μg/ml) for 4 hours. The percentage cholesterol efflux was calculated by dividing the media-derived radioactivity by the sum of the radioactivity in the media and the cells. The effect of HA on the cholesterol efflux was studied, adding that into the medium before the analysis of the cholesterol efflux. Under the existence of medium molecular weight HA (75-350 kDa), the HDL-mediated cholesterol efflux had significantly increased by 10.9% (n=4, p<0.01) in J774.1 macrophages. We also researched the relationship between HA and ATP-binding cassette transporter G1 (ABCG1), because expression of ABCG1 enhances cholesterol efflux to HDL. Predictably, HA significantly increased ABCG1 mRNA expression in macrophage (n=4, p<0.01), determined by Real-time Quantitative RT-PCR. These data suggest that HA increases HDL-mediated cholesterol efflux via ABCG1 in macrophages and the difference of molecular weight on HA may influence ApoA1 and HDL-mediated cholesterol efflux. The relationship between HA and cholesterol efflux might be one of the important mechanisms to prevent progression of atherosclerosis and diabetic macroangiopathy.

Disclosure

K. Matsuki: None. M. Ogura: None. M. Harada-Shiba: Advisory Panel; Self; Aegerion Pharmaceuticals. Speaker's Bureau; Self; Amgen Inc., Astellas Pharma US, Inc., Sanofi, Boehringer Ingelheim Pharmaceuticals, Inc., AstraZeneca, Daiichi Sankyo Company, Limited, MSD K.K.. Research Support; Self; Sanofi, Aegerion Pharmaceuticals, Kowa Pharmaceuticals America, Inc..
Activation of Brown Adipose Tissue Attenuates Diabetic Nephropathy in Mice
Authors: GUAN MEIPING , CAI YINGYING , ZOU SHAOZHOU
Published: June 2018

Brown adipose tissue (BAT) has been regarded as a potential target organ for the treatment of obesity and related metabolic disorders. However, the protective effect of BAT on type 2 diabetic nephropathy (DN) remains unclear. We aimed to test if BAT activation could prevent the development of DN in mice. Type 2 diabetic mice were induced by streptozotocin (STZ) combined with high-fat diet (HFD). CL 316,243, a β3-adrenergic receptor agonist to activate BAT, was administered intraperitoneally (1mg/kg/day) to diabetic mice for 4 weeks. There were no differences in body weight between the groups. Interestingly, the kidney weight and KW/BW ratio were significantly higher in the DM-Con mice compared with the Con and DM+CL mice (p < 0.01). Treatment with CL 316,243 significantly reduced blood glucose and improved dyslipidemia. Simultaneously, activation of BAT led to marked decreases in urinary levels of albumin and 8-hydroxy-deoxyguanosine (8-OH-dG), suppressed fibrotic and inflammatory cytokine synthesis, and finally ameliorated renal morphological abnormalities. In addition to obviously enhancing BAT activity and white adipose tissue (WAT) catabolism, CL 316,243 significantly increased serum adiponectin and the renal sensitivity to FGF-21. Furthermore, the CL 316,243 treatment reactivated the AMPK-SIRT1-PGC1α signaling pathway in kidneys. These findings suggest that activation of BAT attenuates type 2 DN in mice through both the improvement of systemic metabolism and the renoprotective effect of adipokines.

Disclosure

M. Guan: None. Y. Cai: None. S. Zou: None.
On-Demand, App-Delivered Coaching to Enable Diabetes Self-Management in Adults with Type 2 Diabetes—A Feasibility Study
Authors: HERNANDEZ JOHN B. , LEE AMY ARMENTO , ROBERTSON SCOTT , SILVER CARA , MAJITHIA AMIT
Published: June 2018

Introduction: Diabetes self-management training (DSMT) is clinically effective but places a heavy burden on patients. Personalized coaching delivered via scalable mobile app platforms has the potential to provide much needed patient support.

Objectives: This study assessed feasibility and participant experience of a new mobile app-based platform for delivering DSMT to people with type 2 diabetes (T2D). The primary objective was to evaluate participant enrollment (recruitment into the study and completing first sign-in). Engagement with app features and health coach for the study duration was also evaluated.

Methods: Adults with T2D were enrolled into a 4-week prospective, single arm, observational study of the T2D App-based program. The T2D App provided medication reminders, meal logging, blood glucose (BG) tracking, and secure text messaging with a health coach. Participant engagement, satisfaction, participation in self-management behaviors, and finger stick BG results were also measured.

Results: Of 67 participants recruited 99% (66) were able to sign-in to the app and 64 of 67 (96%) completed the 4-week program. The majority of program participants used the T2D App daily (median 100% of days, mean 95% of days). Engagement rates in coach messaging and BG logging exceeded 95% of subjects and were sustained across the study duration. Average blood glucose (AG) in the study participants decreased from week 1 to week 4 (mean -7.8 mg/dL, sd 20.3, P=0.008). The subgroup of individuals with starting AG greater than 154 mg/dL (n=13) experienced the largest declines in AG (mean -29.6 mg/dL, sd 17.9, P<0.0001).

Conclusion: This initial feasibility study of a T2D App-based diabetes management program successfully enrolled participants and sustained their engagement. A decrease in AG was observed which, if sustained, would be clinically significant. Future studies with long term glycemic endpoints and an active control group are needed to validate this observation.

Disclosure

J.B. Hernandez: Employee; Self; Verily Life Sciences LLC.. A. Armento Lee: None. S. Robertson: Employee; Self; Verily Life Sciences LLC. C. Silver: Employee; Self; Verily Life Sciences LLC. A. Majithia: Employee; Self; Onduo.
NADPH Oxidase Nox5 Aggravates Renal Injury in Akita Mouse Model of Diabetic Nephropathy
Authors: JHA JAY C. , DAI AOZHI , COOPER MARK E. , TOUYZ RHIAN M. , KENNEDY CHRIS , JANDELEIT-DAHM KARIN
Published: June 2018

Background: Renal reactive oxygen species (ROS) play an important role in mediating kidney injury in diabetes. Increasing evidence suggests that the pro-oxidant enzyme, Nox5 plays a significant role in human diabetic nephropathy (DN). Nox5 is present in humans and rabbits but not in mice or rats. Thus, there is a paucity of information about Nox5 in conventional animal models of DN. We examined the role of Nox5 in the insulin deficient diabetic Akita mice model using human inducible transgenic mice that express Nox5 selectively in endothelial cells (VEcad+Nox5+) or in mesangial cells (SM22+Nox5+). We also examined the endogenous expression of Nox5 in a high fat fed rabbit model of kidney disease.

Methods: At week 10 mice were culled and kidneys were removed for the assessment of structural damage as well as gene and protein expression of markers of inflammation, fibrosis and oxidative stress. Protein expression of Nox5 and its localization in glomerular cells (endothelial and mesangial cells) were examined in transgenic mice by immunostaining. We also examined expression of pro-fibrotic gene in high fat fed rabbits by next generation sequencing (NGS) and RT-PCR and renal injury by histochemistry.

Results: Expression of Nox5 was confirmed in glomerular endothelial and mesangial cells of transgenic mice. Diabetes induced increase in glomerulosclerosis, gene and protein expression of fibronectin and MCP-1 as well as nitrotyrosine were further increase in both diabetic Nox5 transgenic mice. Moreover, increased expression of Nox5 in high fat fed rabbits vs. normal diet fed rabbits was associated with increased expression of fibronectin, CTGF, collagen IV and VCAM-1 as well as increased mesangial expansion in the kidney.

Conclusions: These findings in both transgenic mice model (endothelial and mesangial cells) suggest that Nox5 plays a significant role in mediating renal injury in diabetes.

Disclosure

J.C. Jha: None. A. Dai: None. M.E. Cooper: Advisory Panel; Self; AbbVie Inc.. Research Support; Self; Boehringer Ingelheim GmbH. Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc.. Speaker's Bureau; Self; Eli Lilly and Company. Advisory Panel; Self; Merck Sharp & Dohme Corp., AstraZeneca. Research Support; Self; Novo Nordisk A/S. Advisory Panel; Self; Novartis AG. R.M. Touyz: None. C. Kennedy: Research Support; Self; Prometic Life Sciences Inc.. K. Jandeleit-Dahm: None.
Early Growth Response 1 (Egr1) Is a Transcriptional Activator of RAAS in Diabetic Kidney Disease
Authors: XUE YAO-MING
Published: June 2018

Background: Egr1 and renin-angiotensin system(RAAS)are invlovled in DKD, but the relationship between them is unclear.

Methods: Egr1 and RAAS were detected in HFD/STZ-induced mice and HK-2 cells treated with TGF-β1. Then, RAAS expression was detected in HK-2 cells and mice with overexpression and knock-down of Egr1. The direct relationship between Egr1 and RAAS was explored via ChIP assay.

Results: We found increased levels of Egr1 and RAAS in the kidney cortices of DKD mice and TGF-β1-treated HK-2 cells. Overexpression or silencing of Egr1 in HK-2 cells and DKD mice could upregulate or downregulate the expression of RAAS. ChIP assays revealed that TGF-β1 induced Egr1 to bind to the RAAS promoter.

Conclusions: Our study provides strong evidence that Egr1 is a transcriptional activator of RAAS in DKD.

Disclosure

Y. Xue: None.
Changes of Plasma and Urine hsa-miR-223-3p Was Correlated with the Severity in Different Stages of Diabetes Kidney Disease of Type 2 Diabetes Patients in China
Authors: HE JUNYI , LI RONG , LI LAN , WU YANAN , YANG QIUPING , ZHANG LIHUA
Published: June 2018

We have previously shown that hsa-miR-223-3p can be as a candidate novel biomarker performing important roles in Diabetes Kidney Disease (DKD) disease process,resulted from co-expression analysis from miRNA (Agilent chips) and lncRNAs (Affymetrix HTA2.0 chips) in plasma.This study involved into 90 patients with type 2 diabetes mellitus (T2DM) and normal control (n=28). Then we divided these diabetes patients into Normoalbuminuric groups(DM) (n=30), Microoalbuminuric group (Micro-DKD) (n=30) and Macroalbuminuric group (Macro-DKD) (n=30), according to urinary albumin to creatinine ratio (ACR) levels (<30mg/g; 30-300mg/g; >300mg/g). The clinical data and biochemical indexes of all the subjects were collected. Real-time qPCR to detect miR-223-3p expression of plasma and urine were reported and compared by using the one-way ANOVA. Statistical analysis was performed using SPSS version 23 software. Receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC) were used to estimate the diagnostic value of the candidate miRs. Compared with controls, the expression of miR-223-3p in plasma were decreased significantly from DM,Micro-DKD to Macro-DKD groups (P <0.001), and the decline was gradually accompanied with the progression of DKD (P <0.001). Same decline pattern was observed in urine expression among four groups, however discrimination was not significant due to the limited patients enrollment in the study. The ROC curve showed that the level of miR-223-3p in plasma can be distinguished among Micro-DKD, Macro-DKD with controls [AUC = 0.750, 95% CI = 0.569-0.923, p = 0.009; AUC = 0.998, 95% CI =0.992-1.000, p = 0.000]. Therefore, plasma hsa-miR-223-3p is a novel biomarker for the severity in Different Stages of DKD in China.

Disclosure

J. He: None. R. Li: None. L. Li: None. Y. Wu: None. Q. Yang: None. L. Zhang: None.
Identifying a New Pathway to Regulate Autophagy in HK2 Cells
Authors: YAMADA EIJIRO , OKADA SHUICHI , YAMADA MASANOBU
Published: June 2018

Diabetic nephropathy (DN) is one of the major diabetic complications, the leading cause of the end stage renal disease (ESRD). Recently autophagy, which is also implicated in diabetes and insulin resistance, was shown to regulate DN. Recently, we demonstrated that Fyn functions as a muscle mass regulator by suppressing macroautophagy through a STAT3-mediated inhibition of the Vps34/Beclin1/ATG14 complex 1 but it is still not known whether Fyn regulates autophagy in kidney and function as a pathogenesis of DN. To investigate this, siRNA mediating knockdown of Fyn in HK2 cells as an in vitro cell model of renal proximal tubular epithelial cells was performed and demonstrating not only decreased p62 expression but also increased LC3-1/2 ratio implicating increased autophagy. To further investigating autophagic flow we utilized ammonium chloride and leupeptin to inhibit the autophagic flow and consistently found that expression of LC3-2 was increased. Phospho-proteomic analysis were performed in vivo and revealed that Fyn phosphorylates Transglutaminase 2 (Tgm2), a known autophagic inhibitor through Beclin1, on Y369 and Y617. To demonstrate that Tgm2 is a direct Fyn substrate, we incubated human purified GST-Tgm2 protein with purified Fyn-CA. In the presence of ATP, Fyn was observed to robustly phosphorylate the Tgm2. Interestingly, Fyn-dependent phosphorylation of Tgm2 was abolished when Y369 and Y617 in Tgm2 was mutated (Tgm2-Y369/617F), suggesting that these sites were specific and unique for Fyn kinase. SiRNA mediating knockdown of Tgm2 in HK2 cells revealed decreased p62 expression and increased LC3-1/2 ratio while overexpressing Tgm2-WT decreased autophagic flow demonstrating Tgm2 could regulate autophagy in the renal tubulus. Taken together, these data suggest that Fyn phorphorylates Tgm2 on Y369 and Y617 and regulates autophagy in the renal tubulus implicating a pathogenesis of DN.

Disclosure

E. Yamada: None. S. Okada: None. M. Yamada: None.
Unbiased Stereological Quantification of Key Features of Diabetic Nephropathy
Authors: PEDERSEN TANJA X. , JOHANSEN THEA T. , FINK LISBETH N. , VRANG NIELS , SECHER THOMAS
Published: June 2018

Diabetic nephropathy (DN) is a serious long-term complication of diabetes. There is no curative treatment of DN, and the lack of knowledge about mechanisms leading to DN hampers the development of efficient therapies. To facilitate drug development, the ability to quantify key histopathological changes in animal models of human DN is essential. In order to develop a more precise determination of translatable nephrotic changes, we used novel and refined image analyses and stereological quantification of key features of DN in a model approach combining genetic diabetes (db/db mice) with uninephrectomy (UNx). UNx or sham-operation was performed in db/db or db/+ male and female mice (n=8-16 mice per group) and the study was terminated 10 or 16 weeks after surgery. Diabetes development was similar in sham and UNx db/db mice. Compared to sham-operation, UNx increased average kidney weight in db/db females both after 10 (43%) and 16 weeks (48%). Blood urea nitrogen (BUN) was increased in UNx vs. sham-operated animals 4 weeks post-surgery, but not at termination. Notably, kidney fibrosis was increased in UNx and sham db/db vs. sham db/+ females (16 weeks; mg total collagen; 12.7 ± 1.3 and 10.7 ± 1.8, respectively, vs. 5.7 ± 0.8, p<0.05). Urinary albumin was progressively increased in all db/db groups with significantly lower variance in UNx db/db females than sham (F-test, p=0.02), permitting smaller group sizes in pre-clinical studies. Unbiased stereological quantification is ongoing to establish volume of glomeruli and number of podocytes. Glomerular collagen is quantified by double-fluorescence imaging concomitantly detecting podocin and collagen.

In conclusion, renal hypertrophy and raised plasma BUN suggest increased pressure on the remaining kidney in UNx mice, leading to fibrosis and decreased kidney function. The thorough quantification of translatable features of DN in the db/db UNx model may provide refinement of this model enabling improved pharmacological testing of compounds targeting DN.

Disclosure

T.X. Pedersen: Employee; Self; Gubra. T.T. Johansen: None. L.N. Fink: Employee; Self; Gubra. Stock/Shareholder; Self; Novo Nordisk A/S. N. Vrang: Board Member; Self; Gubra. Employee; Self; Gubra. Stock/Shareholder; Self; Gubra. T. Secher: Employee; Self; Gubra. Employee; Spouse/Partner; Novo Nordisk A/S.
Apolipoprotein C3 and Cardiovascular Disease in Patients with Type 1 Diabetes and Diabetic Nephropathy
Authors: STECHEMESSER LARS , FORSBLOM CAROL , WEITGASSER RAIMUND , GROOP PER-HENRIK
Published: June 2018

Background and Aims: Apolipoprotein C3 (ApoC3) is a key regulator of triglyceride metabolism via its inhibitory effects on lipolysis and hepatic remnant uptake. Emerging evidence indicate that ApoC3 is an independent risk factor for cardiovascular events. The fact that glucose and insulin regulates ApoC3 expression raises the role of ApoC3 and cardiovascular risk in diabetes. We, therefore, investigated ApoC3 and its association with cardiovascular disease (CVD) in patients with and without diabetic nephropathy.

Methods: This cross-sectional and prospective analysis was part of the prospective, ongoing Finnish Diabetic Nephropathy (FinnDiane) Study. Between 1994 and 2015 data were obtained from 3926 type 1 diabetes (T1D) patients at more than 80 hospitals or health centers across Finland. ApoC3 levels were explored by groups of albuminuria, CKD stages, presence of CVD as well as prediction of CVD and death. Survival curves were calculated by Cox regression analysis.

Results: At baseline, normo-, micro- and macroalbuminuria were present in 71.7%, 13.8% and 14.5% of the population (n=3926, females 48%, age 37.8±12.2 years, diabetes duration 23.2±13 years, HbA1c 8.4±1.5%). CKD stage 3-5 was diagnosed in 14.3%. Coronary heart disease or stroke (CVD) were present in 5.5% at baseline, while 16.3% developed CVD during 15-year follow-up. Compared to normoalbuminuria ApoC3 was elevated in the presence of micro- (p=0.013) or macroalbumiuria (p<0.001). Increasing ApoC3 levels were observed alongside progression of CKD stage (p<0.001). Notably, higher baseline ApoC3 correlated with presence of CVD at baseline, development of CVD during follow-up and death. Differences in survival of those with the highest quartile of ApoC3 were independent of eGFR, triglycerides or HbA1c.

Conclusions: Baseline ApoC3 levels were elevated in T1D patients with micro- and macroalbuminuria, with impaired renal function, and with CVD. ApoC3 also predicted the development of CVD and death during follow-up.

Disclosure

L. Stechemesser: None. C. Forsblom: None. R. Weitgasser: None. P. Groop: Other Relationship; Self; AstraZeneca, AbbVie Inc., Boehringer Ingelheim GmbH, Eli Lilly and Company, Janssen Pharmaceuticals, Inc., Medscape, Merck Sharp & Dohme Corp., Novartis AG, Novo Nordisk A/S, Sanofi.
Recreational Running Training Favourably Affects Antioxidants Gene Expression in Type 2 Diabetes Patients
Authors: SOUSA MAYSA , SANTOS ARITANIA , SILVA MARIA ELIZABETH R.
Published: June 2018

Increased levels of oxidative stress and inflammation have been linked to the progression of diabetes. To reduce oxidative stress and inflammation related to diabetes, diet and chronic exercise is often recommended. Data suggests high intensity interval training may be more beneficial than traditional aerobic exercise. However, appraisals of differing modes of exercise, along with explanations of mechanisms responsible for observed effects in humans, are lacking. This study assessed effects of twelve weeks of two models of high intensity intermittent training, recreational soccer (SO+D) or running (RU+D) training both combined with a caloric restricted diet vs. diet alone (D) in PBMC gene expression of type 2 diabetes patients. Training sessions were performed for 40 minutes, 3 times per week for 12 weeks. We examined PBMC mRNA expression of genes related to antioxidant enzyme activity (CASP9, SOD1, CAT, CASP3). Compared to the D group, only RU+D group resulted in increased mRNA expression of CASP9, SOD1 and CAT (p<0.01) whereas SO+D group mRNA expression did not alter after 12 weeks intervention. Compared to SO+D group, RU+D resulted in increased mRNA expression of CAT, CASP9, SOD1 and CASP3 (p<0.01) post-12 weeks treatment. The combined use of running training associated with dietary advice beneficially influenced expression of genes related to endogenous antioxidant enzyme activity and inflammation than soccer training and diet alone.

In conclusion, recreational running training was superior than soccer training and diet alone elicited lower oxidative stress and inflammation among type 2 diabetes patients.

Disclosure

M. Sousa: None. A. Santos: None. M.R. Silva: None.
Presence of Carotid Artery Plaque Is Associated with Rapid Renal Function Decline in Patients with Type 2 Diabetes and Normal Baseline Renal Function
Authors: SEO DA HEA , KIM SO HUN , AHN SEONG HEE , HONG SEONGBIN , NAM MOONSUK , WOO JEONG-TAEK , BAIK SEI HYUN , LEE KWANWOO , KIM YOUNG SEOL , PARK YONGSOO
Published: June 2018

Background: Recent clinical evidences indicate that early rapid renal function decline is closely associated with the development of diabetic kidney disease. We have investigated the incidence and baseline clinical predictors of rapid renal function decline in patients with type 2 diabetes and normal baseline renal function.

Methods: A total of 967 type 2 diabetic patients from the Korean National Diabetes Program (KNDP), a prospective, multicenter, observational cohort study, with serial GFR measurement for 5 years and normal renal function were included for the analysis. Rapid renal function decline was defined as an eGFR decline >3.3% per year. Carotid ultrasonography was used to assess carotid intima-media thickness (IMT) and the presence of plaque.

Results: The mean age was 53.7 years with body mass index of 25.4 kg/m2. Rapid renal function decline developed in 158 participants (16.3%) and average GFR decline was -5.2 ± 1.9% /year in rapid decliners and -0.4 ± 1.9% /year in non-decliners. There were no differences in the presence of albuminuria or retinopathy between the rapid decliners and non-decliners at baseline. Multivariable logistic regression analyses revealed that female sex (odds ratio (OR) 4.80 [95% CI 2.69-8.56], p<0.0001), higher urine albumin/creatinine ratio (OR 1.26 [1.03-1.55], p=0.024), presence of carotid artery plaque (OR 2.58 [1.47-4.53], p=0.001), and previous history of hypertension (OR 1.93 [1.09-3.44], p=0.025) and CVD (OR 2.53 [1.24-5.15], p=0.011) were the independent clinical predictors for rapid renal function decline.

Conclusions: Our study suggests the need for close monitoring of renal function and early intensive management in patients with type 2 diabetes and carotid atherosclerosis.

Disclosure

D. Seo: None. S. Kim: None. S. Ahn: None. S. Hong: None. M. Nam: None. J. Woo: None. S. Baik: None. K. Lee: None. Y. Kim: None. Y. Park: None.
Serum Adipocyte Fatty Acid-Binding Protein Levels Are Linked to Rapid Renal Function Decline in Patients with Type 2 Diabetes and Normal Renal Function
Authors: SEO DA HEA , KIM SO HUN , AHN SEONG HEE , HONG SEONGBIN , NAM MOONSUK
Published: June 2018

Background: Rapid renal function decline has been recognized as an important predictor for diabetic nephropathy (DN). Adipocyte fatty acid-binding protein (A-FABP) is a major cytoplasmic protein in adipocytes and is closely associated with metabolic syndrome and type 2 diabetes. We aim to investigate the role of adipokines including A-FABP and inflammatory cytokines in the development of rapid renal function decline in type 2 diabetes and normal renal function.

Methods: A total of 456 subjects with type 2 diabetes with normal renal function were recruited from an outpatient clinic at the Diabetes Center of Inha University Hospital and were followed up for 6 years with serial glomerular filtration rate (GFR) measurements. The serum levels of tumor necrosis factor (TNF)-a, and interleukin (IL)-6, high molecular weight (HMW) adiponectin, endogenous secretory receptor for advanced glycation end products (esRAGE), A-FABP and pentraxin-related protein (PTX3) were measured at baseline. Rapid renal function decline was defined as an eGFR decline >3.3% per year.

Results: During follow-up, 120 participants (26.3%) developed rapid renal function decline and mean annual eGFR decline was -5.4 ± 2.7% and -0.6 ± 2.4% in decliners and non-decliners, respectively. Median A-FABP levels were significantly higher in patients with rapid decliners than in non-decliners (19.1 vs. 16.8 ng/ml, p=0.011). No significant difference of three cytokines levels were observed between groups. Each log unit increase of A-FABP was independently associated with greater risk of rapid renal function decline (odds ratio [OR] = 3.60; 95% confidence interval [CI] 1.95-6.66; p < 0.001) after adjustments for sex, BMI, A1C, GFR, hypertension, presence of carotid artery plaque and urine albumin creatinine ratio.

Conclusions: High plasma A-FABP was an independent risk factor for rapid renal function decline in type 2 diabetes and normal renal function.

Disclosure

D. Seo: None. S. Kim: None. S. Ahn: None. S. Hong: None. M. Nam: None.