bluuebunny/crossref_metadata_2025_split huggingface dataset:
47-OR: Regulation of SIRT1 as a Target of Prevention of Diabetic Retinopathy in db/db Mice
Authors: VIEIRA CRISTIANO P. , LONGHINI ANA LEDA , CALZI SERGIO LI , DESIREE` DUPONT MARIANA , BUSIK JULIA V. , GRANT MARIA B. 
Published: June 2019

SIRT1 is critical to vascular health and regulates glucose and lipid metabolism through its deacetylase activity. Type 2 diabetes-associated decreases in SIRT1 levels result in increased levels of acetylated LXR leading to reduced expression of reverse cholesterol transport genes and increased expression of NFκB-controlled pro-inflammatory genes. Diabetic circulating angiogenic cells (CAC) which are critical in vascular repair also demonstrate reduced SIRT1 and increasing its levels corrects CAC dysfunction (1). db/db mice were given SRT-1720 hydrochloride (100 mg/kg body weigh/day for 2 or 6 mo.) in their chow or fed normal chow beginning at diabetes onset. Retinal tissue, blood and bone marrow (BM) were collected. Electroretinograms (ERG) were performed at 2 and 6 mo. The BM supernatant was analyzed for cytokines. BM hematopoietic stem cells and CACs were assessed by flow cytometry. Retinal lipids were assessed by Mass Spectroscopy. At 2 mo, the retina of SRT1720-treated db/db mice demonstrated an increase in the ratio of total cholesterol/demosterol/lanosterol (197±85) compared to db/db (65±12. p&lt;0.05). The percent of CAC was increased in db/db mice treated with SRT1720 (2.1±0.6 vs. 0.2±0.1 db/db. p&lt;0.01). Most interestingly, SRT1720-treated db/db mice showed a decrease in the percentage of proinflammatory granulocyte-macrophage progenitors (1.7±0.3 vs. 2.8±0.6 db/db, p&lt;0.008) and BM IL-1 beta (5±0.5 SIRT vs. 10±2.4 control db/db, p&lt;0.01) levels. db/db mice with 6 months of diabetes showed a significant reduction in the ERG scotopic response and SRT-1720 treatment corrected this dysfunction (165±60 in SRT1720 db/db vs. 76±20µV, p&lt;0.03), compared to controls (194±60µV). These findings support that activation of SIRT1 has multiple LXR-mediated beneficial effects. Namely, it improved the visual response in T2D model by regulating retinal cholesterol metabolism, increased vascular protective CACs and decreased pro-inflammatory progenitor cells and cytokines in the BM.

Disclosure
C.P. Vieira: None. A. Longhini: None. S. Li Calzi: None. M.D. DuPont: None. J.V. Busik: None. M.B. Grant: None.

Funding
National Institutes of Health (EY025383)
479-P: Combination of Trans-resveratrol and Hesperetin Corrects Unscheduled Increased Glucose Metabolism Driving Dysfunction of Human Periodontal Ligament Fibroblasts in Hyperglycemia In Vitro
Authors: RABBANI NAILA , ASHOUR AMAL A. , XUE MINGZHAN , THORNALLEY PAUL 
Published: June 2019

Dysfunction of human periodontal ligament fibroblasts (PDLFs) contributes to increased prevalence and severity of periodontitis in diabetes. Hyperglycemia induces multiple pathways of metabolic dysfunction: mitochondrial dysfunction, hexosamine and protein kinase C pathway activation and increased formation of methylglyoxal (MG) in human periodontal ligament fibroblasts in vitro. We recently showed that increased glucose metabolism driving metabolic dysfunction in hyperglycemia is produced by glucose-induced stabilization of hexokinase-2 (HK2) to proteolysis, increasing HK2 activity. In this study we investigated correction of this by combination of resveratrol and hesperetin (tRES-HESP) which induces expression of glyoxalase 1 (Glo1) and protects against increased MG. Human PDLFs were incubated in primary culture with 8 mM or 25 mM glucose for 3 days with and without 10 µM tRES-HESP. Glucose consumption and flux of MG formation was measured by enzymatic assay. Activity of glyoxalase 1 (Glo1) was measured by spectrophotometric assay. HK2 protein was determined by Western blotting. The flux of glucose consumption in high glucose concentration cultures of PDLFs was increased, compared to low glucose concentration control (10,658 ± 1,141 vs. 5,923 ± 706 nmol/106 cells/day, n = 3; +80%, P&lt;0.01). Flux of formation of MG was increased similarly: 9.74 ± 0.42 v 6.79 ± 0.47 nmol/106 cells/day, n = 3; +44%, P&lt;0.001). Glo1 activity was decreased in high glucose culture: 485 ± 78 vs. 946 ± 99 mU/mg protein (-49%, P&lt;0.001). HK2 protein was increased: +32% at day 3 (P&lt;0.05). tRES-HESP corrected increased glucose consumption and MG formation, and decreased Glo1 activity in high glucose concentration cultures. tRES-HESP offers a new therapy to prevent diabetic periodontitis.

Disclosure
N. Rabbani: None. A.A. Ashour: None. M. Xue: None. P. Thornalley: None.
485-P: Combination of Trans-resveratrol and Hesperetin Corrects Unscheduled Increased Glucose Metabolism Driving Endothelial Cell Dysfunction in Hyperglycemia In Vitro
Authors: THORNALLEY PAUL , ASHOUR AMAL A. , XUE MINGZHAN , RABBANI NAILA 
Published: June 2019

Endothelial dysfunction contributes to the development of vascular complications of diabetes. Hyperglycemia induces multiple pathways of metabolic dysfunction: mitochondrial dysfunction, hexosamine and protein kinase C pathway activation and increased formation of methylglyoxal (MG). We recently showed that increased glucose metabolism driving this is produced by glucose-induced stabilization of hexokinase-2 (HK2) to proteolysis, increasing HK2 activity. In this study we investigated correction of this by combination of resveratrol and hesperetin (tRES-HESP) which induces expression of glyoxalase 1 (Glo1) and protects against increased MG. Human aortal endothelial cells (HAECs) were incubated in primary culture with 5 mM or 20 mM glucose for 6 days with and without 10 µM tRES-HESP. Glucose consumption and flux of MG formation was measured by enzymatic assay. Activity of glyoxalase 1 (Glo1) was measured by spectrophotometric assay. HK2 protein was determined by Western blotting. The flux of glucose consumption in high glucose concentration cultures of HAECs was increased, compared to low glucose concentration control (1647 ± 242 vs. 748 ± 150 nmol/106 cells/day, n = 8; +120%, P&lt;0.01). Flux of formation of MG was increased similarly: 1.13 ± 0.10 v 0.62 ± 0.07 nmol/106 cells/day, n = 8; +81%, P&lt;0.01). Glo1 activity was decreased in high glucose culture: 1822 ± 143 vs. 1370 ± 160 mU/mg protein (-25%, P&lt;0.01). HK2 protein was increased: +94% at day 3 and + 58% at day 6 (P&lt;0.001). tRES-HESP corrected increased glucose consumption and MG formation, and decreased Glo1 activity in high glucose concentration cultures. This was achieved by inducing expression of glucose-6-phosphate dehydrogenase (G6PD), decreasing HK2 expression by decreasing the glucose-6-phosphate (G6P) and related carbohydrate response element transactivation by G6P/Mondo A/Mlx. tRES-HESP offers a new effective therapy of endothelial dysfunction.

Disclosure
P. Thornalley: None. A.A. Ashour: None. M. Xue: None. N. Rabbani: None.
49-OR: Association of Gene Polymorphisms and Plasma MiRNAs with Diabetic Retinopathy
Authors: POLINA EVELISE REGINA , SBRUZZI RENAN C. , DE OLIVEIRA FERNANDO MARQUES , CRISPIM DAISY , ENOIA MARIA ENOIA D. , CANANI LUIS H. , SANTOS KATIA G. 
Published: June 2019

Circulating microRNAs (miRNAs/miRs) have emerged as potential disease biomarkers; however, their expression in diabetic retinopathy (DR) has been evaluated only recently in humans. Whilst it is recognized that sequence variants in miRNA genes (miRSNPs) can alter the miRNA-mediated regulation of gene expression, there are almost no studies on the association of miRSNPs with DR. In this case-control study, we investigated whether the functional polymorphisms rs531564 G&gt;C (MIR124-1), rs4636297 A&gt;G (MIR126) and 2910164 C&gt;G (MIR146A) are associated with DR in 1,025 Brazilians with type 2 diabetes. We also evaluated whether the plasma levels of miR-124, miR-126 and miR-146a are associated with DR and with miRSNPs in 182 patients. Blood donors were included to determine the prevalence of miRSNPs (n = 104) and miRNA expression (n = 20) in the general population. Genotyping was done by real-time PCR and plasma levels were quantified by RT-qPCR. Genotype and allele frequencies in diabetic subjects did not significantly differ from those of blood donors, and the minor allele frequencies were similar to those reported in other populations. The MIR126 heterozygous genotype was more frequent in patients without DR than in those with DR (52% versus 44%, respectively, p = 0.040), and remained associated with this complication after controlling for clinical covariates. MiR-126 was markedly downregulated in diabetic subjects as compared to blood donors and even more reduced among those with DR (mean log2 fold-change of -0.7 ± 1.7, -1.1 ± 2.0 and -1.7 ± 1.6 for subjects without DR, with nonproliferative DR or with proliferative DR, p = 0.002). Patients carrying the A allele of the rs4636297 polymorphism had almost twice higher levels of miR-126 than patients with GG genotype. MiR-126 and miR-146a were strongly correlated and the levels of miR-146a were also lower in diabetic subjects than in blood donors. Overall, our findings reinforce that gene variants and expression of miRNAs are associated with DR in type 2 diabetes.

Disclosure
E.R. Polina: None. R.C. Sbruzzi: None. F.M. de Oliveira: None. D. Crispim: None. M.D. Enoia: None. L.H. Canani: Research Support; Self; Novo Nordisk Inc. Speaker's Bureau; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi. K.G. Santos: None.

Funding
Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (1994-2551/13-4); Fundo de Incentivo à Pesquisa e Eventos do Hospital de Clínicas de Porto Alegre (14-0645)
492-P: A Novel Renoprotective Target of SGLT2 Inhibitor: HIF-1α Inhibition and AMPK Activation in Renal Proximal Tubular Epithelial Cells
Authors: BESSHO RYOICHI , TAKIYAMA YUMI , TAKIYAMA TAKAO , OTA TSUGUHITO 
Published: June 2019

The renoprotective mechanism of SGLT2 inhibitors has not been fully elucidated. Tubular hypoxia is major driving force for proximal tubulopathy in diabetic kidney. In addition, diabetes causes dysregulation of AMP-activated protein kinase (AMPK) in kidney cortex. In this study, we assessed the effects of luseogliflozin, an SGLT2 inhibitor, on hypoxia inducible factor-1α (HIF-1α) expression and AMPK phosphorylation in cultured human renal proximal tubular epithelial cells (HRPTECs) and on tubulointerstitial pathological changes in diabetic mice. Luseogliflozin inhibited hypoxia (1%O2, 24h)-induced HIF-1α protein expression in HRPTECs in a dose-dependent manner (1-100µM). In addition, luseogliflozin increased AMPKα protein phosphorylation (Th172) in normoxic and hypoxic condition. Intriguingly, luseogliflozin suppressed oxygen consumption rate (OCR) measured by oxygen quenching phosphorescent probe by 69% in HRPTECs from normoxic condition. Hypoxia decreased OCR by 52% and luseogliflozin further decreased OCR by 33% from hypoxic condition. Moreover, luseogliflozin rescued hypoxic state in HRPTECs even under hypoxic conditions assessed by a hypoxia-sensitive dye, pimonidazole. These data indicate luseogliflozin inhibits HIF-1α expression through suppression of mitochondrial OCR, which leads to restore intracellular hypoxia. We next treated db/db mice with 15 mg/kg/day luseogliflozin for 8 weeks. Luseogliflozin ameliorated tubular injury compared to non-treated db/db mice. Furthermore, immunostaining of HIF-1α and fibronectin in cortical tubules revealed that luseogliflozin decreased HIF-1α and fibronectin expression in db/db mice.
In conclusion, luseogliflozin decreases mitochondrial OCR and ameliorates tubular fibrosis at least partly by suppressing HIF-1α expression and activating AMPK in renal proximal tubules of diabetic mice. These results may provide a novel renoprotective target of SGLT2 inhibitor.

Disclosure
R. Bessho: None. Y. Takiyama: None. T. Takiyama: None. T. Ota: None.
496-P: A New Method to Image Sweet Taste Receptors in the Pathogenesis of Diabetic Nephropathy: Detection of 18F Lactisole with PET/CT
Authors: ZHOU LUPING , HUANG WEI , LIU NAN , MA XIU M. , GUO MAN , CHEN QING , GAO CHENLIN , XU YONG 
Published: June 2019

Previous studies showed that the expression of sweet taste receptors (STRs) was obviously downregulated in the pathogenesis of diabetic nephropathy (DN), suggesting that STRs may act as new therapeutic targets of DN. However, STRs are weakly expressed in kidneys, it is necessary to find a non-invasive method for detection of STRs distribution in the whole body and kidney. Lactisole, a broad-acting sweet antagonist, is commercially available and may be useful to assess the function and physiological role of the STRs. Whether the detection of isotope 18F labeled lactisole (18F-lactisole) with PET/CT can be used as a probe for imaging STRs expression differences is unclear. In this study, DN mouse model was induced by streptozotocin (STZ) in vivo, mouse glomerular mesangial cells (GMCs) and human proximal tubular cells (HK-2) were stimulated by high glucose in vitro, exogenous lactisole was used as an intervention reagent, 18F-lactisole was used as probe agent to evaluate the expression and distribution of STRs in the pathogenesis of DN. Our results showed that both the expression and distribution of STRs in mice were clearly imaged by PET/CT via intravenous injection of 18F-lactisole, 18F-lactisole signal in renal tissue was attenuated under the condition of diabetes, this trend persisted 2 hours after furosemide injection. Furthermore, exogenous lactisole significantly mitigated the effect of high glucose on STRs in vitro and vivo, which were detected by PET/CT or radioactive dosimeter via 18F-lactisole. These combined results support the hypothesis that detection of 18F-lactisole with PET/CT can be used as a new probe to image STRs and distinguish its expression level, suggesting a new method for non-invasive diagnosis of DN.

Disclosure
L. Zhou: None. W. Huang: None. N. Liu: None. X.M. Ma: None. M. Guo: None. Q. Chen: None. C. Gao: None. Y. Xu: None.
50-LB: Impact of Diabetes Self-Management Education (DSME) on A1C in High-Risk and Low-Income Chinese Immigrant Populations Residing in Urban Communities
Authors: SITU ESTHER , WOO JOSEPH , ZHANG XINYU 
Published: June 2019

Background: Asian American Medical Group serves predominately monolingual senior or low income Chinese immigrants in the San Francisco Bay Area. This clinical study evaluated the difference of A1c improvement between T2DM patients who received DSME along with standard care vs. standard care alone. DSME has been widely underutilized by physicians in the independent practice setting.
Method: In this study, treatment was applied to 36 patients from a sample of 130 with T2DM with A1c &gt;=8% and the average age were 70 years old for both cohorts per electronic medical record from 2017-2018. Before treatment, the treatment group’s average A1c was 9.83% vs. the control group was 8.96%. A dietitian with CDE was assigned to provide 1:1 counseling to patients/patient’s caregiver(s) on diabetes management, implement patient goals, and provide individualized care. RD actively communicated with PCPs for changes in care as needed. This program was provided to patients based on their demands and needs; ranging from 1-5 visits within a year.
Results: A total of 84% of patients receiving DSME reduced their A1c levels; 68% reducing A1c by &gt;= 0.88%, 41% reducing A1c by &gt;=1.76%. Comparatively, 48% of patients who didn’t receive DSME reduced their A1c levels, 37% reducing A1c by &gt;= 0.88% and 25% reducing A1c by &gt;=1.76%. A comparison between the cohorts that experienced an A1c reduction of &gt;=0.88% revealed statistical significance at 95% confidence level (p = 0.0474). The data shows that there is a 31% difference between these two cohorts. Overall, patients receiving treatment experienced a mean reduction in A1c of 1.54% compared to an increase of 0.08% for the control group. Furthermore, 50% from the treatment side did lower A1c &lt;8% whereas only 26% from the control.
Conclusion: The study demonstrated that DSME program improves A1c and points to the value of DSME. Thus, DSME should be promoted and widely utilized as a standard care regiment in the out-patient setting.

Disclosure
E. Situ: None. J. Woo: None. X. Zhang: None.
513-P: Adenosine Signal Plays an Important Role in Renoprotective Effects of SGLT2 Inhibitor in Proteinuric Diabetic Mice
Authors: SHIMADA KEIJI , TAKAGAKI YUTA , KANASAKI KEIZO , KOYA DAISUKE 
Published: June 2019

Background: SGLT2 inhibitor (SGLT2i) displayed remarkable and early renal protection in overt nephropathy cases in clinical trials. However, detailed molecular mechanisms by which SGLT2i ameliorated renal damage in proteinuric diabetic patients were not elucidated yet. Here, we investigated that SGLT2i exerted renal protective effects via adenosine signal in diabetic mice with overt proteinuria.
Methods: STZ (200mg/kgBW) was injected intraperitoneally to induce diabetes in 8-week-old CD-1 mice. Four weeks later, mice were treated with intraperitoneal injection of bovine serum albumin (BSA: 0.3g/30gBW) with or without oral administration of SGLT2i TA-1887 (30mg/kg).
Result: In the STZ+BSA, pronounced renal tubular damages and interstitial fibrosis were observed; TA-1887 significantly ameliorated. The mRNA microarray analysis of kidney samples revealed that gene expressions related to TGF-β signaling pathway and epithelial-mesenchymal-transition program were increased in the STZ+BSA compared to the Control+BSA; TA-1887 suppressed them. Furthermore, the expression of ecto 5'-nucleotidase (5'-NT), which is essential for production of adenosine from AMP, was suppressed in STZ+BSA compared to Control+BSA; TA-1887 restored 5’-NT levels. In the immunohistological analysis, 5'-NT was expressed in glomeruli including mesangial cells and renal tubules of cortex/medulla in control kidney. However, 5’-NT was suppressed in all areas of STZ+BSA kidney; TA-1887 reversed the 5'-NT expression and distribution. Glomerular filtration rate (GFR) evaluated by inulin clearance was significantly greater in the STZ+BSA group and suppressed by TA-1887; the administration of the adenosine type 1 receptor inhibitor, DPCPX, cancelled the effects of TA-1887 on GFR and renal tubule damage.
Conclusion: The adenosine signal plays an important role in the renal protective effects of SGLT2i in diabetic mice with proteinuria.

Disclosure
K. Shimada: None. Y. Takagaki: None. K. Kanasaki: Other Relationship; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd., Sanofi, Taisho Toyama Pharmaceutical. D. Koya: Research Support; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Mitsubishi Tanabe Pharma Corporation. Speaker's Bureau; Self; Astellas Pharma Inc., Astellas Pharma Inc., AstraZeneca, Merck & Co., Inc., Mitsubishi Tanabe Pharma Corporation, Taisho Pharmaceutical Co., Ltd.

Funding
Japan Society for the Promotion of Science
516-P: Preventive Effect of Sulforaphane on Type 2 Diabetes-Induced Diabetic Nephropathy via AMPK-Mediated Activation of Glucose/Lipid Metabolism and Nrf2 Antioxidative Function
Authors: LI ZHUO , CAI LU , SUN YIKE 
Published: June 2019

AMP-activated protein kinase (AMPK), particularly AMPK-α2 isoform, can attenuate the nephropathic pathogenesis by improving lipid metabolism. The objective of the present study was to define if sulforaphane (SFN) can protect type 2 diabetes (T2D)-induced nephropathy (DN) by activating AMPKα2 and Nrf2-mediated anti-oxidative pathways. AMPKα2 knockout (AMPKα2-KO) mice and their wild type (WT, C57BL/6J) mice were fed high-fat diet (HFD) for 3 months to induce insulin resistance, followed by an i.p. injection of streptozotocin (STZ) to induce hyperglycemia, i.e.:T2D. Control mice were fed normal diet without STZ. T2D and control mice were treated with or without SFN for 3 months and then stop SNF treatment, remained feeding either ND or HFD. These mice were collected 24-hour urine and then sacrificed at 3 and 6 months after diabetes onset to collect kidney’s tissue for pathological examination and molecular assays. Proteinuria was significantly increased in T2D mice at 3 and 6 months, but not in SFN-treated T2D mice. The destruction of kidney structure induced by T2D, including renal fibrosis, lipotoxicity, inflammation, oxidative stress/damage along with significant suppression of AMPK signaling pathways was prevented by SFN in WT diabetic mice. Specifically, T2D-induced, time-dependent from 3 to 6 months, increased accumulation of FN and COL-1, increased expression of TGF-β PAI-1, TNF-α and decreased phosphorylation of AMPKα and its downstream markers (ACC, CPT1-A, Sirt-1, PGC1α, PPARα, and Nrf2). All these pathogenic changes were prevented by SFN in WT diabetic mice, which was more at 6 months compared to 3 months. However, in AMPKα2-KO mice, suggesting that all these are AMPK-dependent. Therefore, the present study suggests that SFN protects against T2D-induced DN via AMPKα2-mediated improvement of lipid metabolisms and up-regulation of Nrf2-mediated anti-oxidative defense system.

Disclosure
Z. Li: None. L. Cai: None. Y. Sun: None.
519-P: Acute Kidney Injury Sensitivity in Diabetic Murine Model
Authors: SONG SANG HEON , KIM SANG SOO , KIM IN JOO 
Published: June 2019

Background: We recently reported that diabetes can upregulate two discrete isoforms of the MMP-2 (full length and N-terminal truncated, FL-MMP-2, NTT-MMP-2) in renal tubules and especially, inducible NTT-MMP-2 could be related to renal tubular damage. Also, pyrrolidinedithiocarbamate (PDTC), NF-Kb inhibitor, can suppress NTT-MMP-2 selectively. This study was conducted to analyze the effects of acute kidney injury on diabetic animal model and test the effect of PDTC in acute kidney injury with diabetes.
Methods: We performed unilateral ischemia-reperfusion injury (uIRI) in streptozotocin-induced diabetic animals as a model of acute kidney injury and followed for 48 hours and six weeks. Mice were randomized into four groups as follows; control (A), non-DM+uIRI (B), DM+uIRI (C), DM+uIRI+PDTC (D).
Results: In a 48-hour experiment, tubular necrosis was more severe in diabetic IR kidney compared with nondiabetic IR kidney. Outer medulla in group C was becoming devastated and led to the extensive coagulative necrosis. Those findings were diminished by PDTC. Peritubular capillary density was increased by PDTC and cortical macrophage density was decreased by PDTC compared to group B and C. The nitrotyrosin was lesser expressed by PDTC compared to group B and C. In a six-week experiment, IRI caused the kidney to be stiffen and shrink both in group B and C and PDTC can ameliorate these gross changes compared to group C. Peritubular capillary density was increased by PDTC compared to group C. Cortical macrophage density was higher than group B and D and PDTC ameliorate the recruitment of macrophage in renal cortex. The expression of cleaved caspase-3/caspase 3 in group C was higher than group B and D.
Conclusion: Ischemia-reperfusion injury caused more intense renal damage in diabetic murine model and could lead to chronic renal change related to inflammation and apoptotic pathway. These changes may be mitigated by PDTC treatment associated with anti-oxidative mechanism and inhibition of NTT-MMP-2.

Disclosure
S. Song: None. S. Kim: None. I. Kim: None.

Funding
National Research Foundation of Korea (2016R1A2B4008243)