bluuebunny/crossref_metadata_2025_split huggingface dataset:
442-P: Comparative Study of the Effects of GLP-1 Analog and SGLT2 Inhibitor against Diabetic Cardiomyopathy in Type 2 Diabetic Rats: Possible Underlying Mechanisms
Authors: EID ELSAYED A. , HUSSEIN ABDELAZIZ , LASHIN LASHIN , TAHA MEDHAT 
Published: June 2020

Objectives: In the present study we investigated the possible cardioprotective effects of GLP-1 and SGLTi against diabetic cardiomyopathy (DCM) in type 2 diabetic rats and the mechanisms underlying these effects.
Methods: 32- male Sprague Dawley rats were randomly subdivided into 4 equal groups; a) control group, b) DM group, type 2 diabetic rats received 2ml oral saline daily for 4 weeks, c) DM+ GLP-1, type 2 diabetic rats were treated with GLP-1 analogue (liraglutide) at a dose of 75 µg/kg via S.C. injection for 4 weeks and d) DM+ SGLTi: type 2 diabetic rats received SGLT2 inhibitor (dapagliflozin) at a dose of 1mg/kg via oral gastric gavage. By the end of treatment (4 weeks), serum blood glucose, HOMA-IR, insulin and cardiac enzymes (LDH, CK-MB) were measured. Also, the cardiac histopathology, myocardial oxidative stress markers (MDA, GSH and CAT) and norepinephrine (NE), myocardial fibrosis, the expression of caspase-3, TGF-β, TNF-α and tyrosine hydroxylase (TH) in myocardial tissues were measured.
Results: T2DM caused significant increase in serum glucose, HOMA-IR index, serum CK-MB and LDH (p&lt; 0.05). This was associated with significant myocardial damage and fibrosis, elevation of myocardial MDA, NE with upregulation of myocardial caspase-3, TNF-α, TGF-β and TH and significant decrease in serum insulin and myocardial GSH and CAT (p&lt;0.05). Administration of either GLP-1 analog or SGLTi caused in significant improvement in all studied parameters (p&lt; 0.05).
Conclusions: GLP-1 and SGLTi exhibited cardioprotective effects against DCM in type 2 diabetic rats. SGLTi showed more cardioprotective effect than GLP-1. The cardioprotective actions of these agents might be due to attenuation of fibrosis, oxidative stress, apoptosis (caspase-3), sympathetic nerve activity and inflammatory cytokines (TNF-α and TGF-β).

Disclosure
E.A. Eid: None. A. Hussein: None. L. Lashin: None. M. Taha: None.
446-P: Skin Autofluorescence, a Measure of Advanced Glycation End Products, and Diabetes Complications: Meta-analysis and Systematic Review
Authors: KOUIDRAT YOUSSEF 
Published: June 2020

Background: Chronic accumulation of advanced glycation end products (AGEs) has been suggested as predictor of cardiovascular complications in diabetes mellitus. Tissue AGEs can be measured noninvasively and reproducibly in the skin by Skin autofluorescence (SAF), a clinical tool developed in the 2000s. This study aimed to estimate the relationship between AGEs indicated by SAF levels and the risk of micro- and macro-vascular complications based on data from observational studies.
Methods: A systematic literature search was performed using PubMed, Embase, and the Web of Science databases from their inceptions until December 2018. Two reviewers identified eligible studies that reported the measure of AGEs by SAF in patients with diabetes mellitus. Meta-analyses using random effects models were used to analyze the data.
Results: Twenty-four published studies involving 2715 patients with diabetes (1756 T2D and 959 T1D), 1673 controls with uncomplicated diabetes and 1283 healthy controls were included in the meta-analysis. Five diabetes-related microvascular complications: Nephropathy, Neuropathy, Retinopathy, Erectile dysfunction and Diabetic foot ulcer and 3 main macrovascular complications : Peripheral arterial diseases, Stroke and CVD were identified. Both, in patients with T1D or T2D, SAF levels were significantly higher compared to healthy controls with substantial heterogeneity (I2=78% in T1D and I2=91% in T2D; P&lt;0.0001). Interestingly, patients with complicated diabetes exhibited significantly higher SAF levels compared to patients with uncomplicated diabetes: for T2D: mean difference [MD]: 0.303 AU; 95% CI: 0.237-0.369) with mild heterogeneity (I2=20%; P&lt;0.0001). For T1D: MD: 0.385AU; 95% CI: 0.289-0.480), with moderate heterogeneity (I2=54%; P&lt;0.0001).
Conclusions: Our findings suggest that SAF measure may play an important role as a biomarker for patients with high risk of diabetes-related vascular complications.

Disclosure
Y. Kouidrat: None.
449-P: Impact of Body Mass Index (BMI) and Waist Circumference (WC) on Coronary Artery Disease (CAD) in Japanese with and without Diabetes Mellitus (DM)
Authors: KOMATSU TAKESHI , FUJIHARA KAZUYA , YAMADA MAYUKO H. , SATO TAKAAKI , KITAZAWA MASARU , YAMAMOTO MASAHIKO , SEIDA HIROYASU , YAMANAKA NAUTA , MATSUBAYASHI YASUHIRO , YAMADA TAKAHO , SONE HIROHITO 
Published: June 2020

BMI ≥25.0 and WC ≥90 are components of the International Diabetes Federation (IDF) definition of metabolic syndrome for Asians. We have investigated more suitable BMI and WC levels for risk of CAD using large-scale nationwide claims data on people with and without DM in the same cohort. The sample included 157,220 Japanese men 18-72 years of age without a known history of CAD at baseline (DM: 15,465 patients, non-DM: 141,755 patients). Multivariate Cox regression model examined the association of BMI/WC categories and their combination with CAD. During the study period (median follow-up 5.3 y), 662 and 332 CAD events occurred in non-DM and DM, respectively. Cox regression model showed that BMI ≥22.5 and WC ≥80 conferred a 1.29 (1.07-1.55), and 1.23 (1.00-1.50) times increased risk of CAD, respectively, in non-DM but that those relationships disappeared in people with DM (HR, 0.96 (0.71-1.29) for BMI and 1.13 (0.78-1.62) for WC. Stratified analysis of 4 groups of combinations of BMI and WC showed that in non-DM with BMI ≥22.5/WC ≥80 the risk of future CAD was significantly increased compared with non-DM with BMI&lt;22.5/WC&lt;80 (Table). Each DM group had an approximately 3-fold increase in CAD events (Table).
In summary, lower BMI and WC values can predict CAD in individuals without DM whereas their influence was modest in people with DM.

Disclosure
T. Komatsu: None. K. Fujihara: None. M.H. Yamada: None. T. Sato: None. M. Kitazawa: None. M. Yamamoto: None. H. Seida: None. N. Yamanaka: None. Y. Matsubayashi: None. T. Yamada: None. H. Sone: Research Support; Self; Kyowa Hakko Kirin Co., Ltd., Novartis AG, Ono Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Co.
452-P: Exercise-Induced Elevation of MicroRNA-181b Ameliorates Endothelial Dysfunction in Diabetic Mice
Authors: SHANG WENBIN , WANG YU , LUO JIANG-YUN , TIAN XIAO YU , WANG LI , HUANG YU 
Published: June 2020

Endothelial dysfunction plays an essential role in the pathogenesis of diabetic vascular diseases. Our previous studies showed exercise improves endothelial function in diabetic mice partially through AMPK pathway. MicroRNA 181b (miR-181b) was reported to inhibit NF-κB activity in endothelial cells. However, it is still unknown whether miR-181b is regulated by AMPK signaling and involved in the vasoprotective effect of exercise. The present study investigated the expression of miR-181b in aortas from diabetic mice with/without exercise and reveals a role of the AMPK in exercise-induced miR-181b expression. We first determined the miR-181b level in the aortas of diabetic db/db mice subjected to treadmill exercise (45 min/day for eight weeks). The results showed that exercise significantly increased miR-181b level in the endothelium of mouse aorta compared to the sedentary mice. To test whether the increased blood flow during exercise affects miR-181b expression, we tested the effect of laminar flow on miR-181b expression in Human Umbilical Vein Endothelial Cells (HUVECs). Laminar flow greatly increases miR-181b expression. To understand the mechanism for flow-induced miR-181b expression, we treated HUVECs with AMPK agonist 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) and found that AICAR-induced miR-181b expression is reversed by AMPK inhibitors. Furthermore, we infected db/db mice with the adenovirus over-expressing miR-181b. MiR-181b overexpression improved endothelial function in db/db mice accompanied by decreased vascular inflammation and increased eNOS expression in mouse arteries. In summary, exercise increases miR-181b level in mouse endothelium probably via shear stress-induced AMPK activation. MiR-181b improves endothelial function in diabetic mice by inhibiting vascular inflammation and increasing eNOS expression. The present results suggest the therapeutic potential of targeting miR-181b against diabetic vasculopathy.

Disclosure
W. Shang: None. Y. Wang: None. J. Luo: None. X. Tian: None. L. Wang: None. Y. Huang: None.

Funding
Hong Kong Research Grants Council/Collaborative Research Fund (C4024-16W); Hong Kong Health and Medical Research Fund (05161746)
455-P: Pan-ErbB Inhibition Attenuates Diabetes-Induced Vascular Dysfunction in an Experimental Model of Diabetes
Authors: AKHTAR SAGHIR , CHANDRASEKHAR BINDU , BENTER IBRAHIM 
Published: June 2020

The underlying mechanisms leading to diabetes-induced vascular complications are not fully understood. We have previously shown that epidermal growth factor receptor (EGFR), which is part of the four membered family of EGFR/ErbB receptor tyrosine kinases, appears to play a detrimental role. However, the role of the other ErbB family members (ErbB2, ErbB3, and ErbB4)- that regulate cell growth, proliferation, differentiation and apoptosis- is not fully known and was further evaluated in this study. In a streptozotocin-rat model of type 1 diabetes, 4-weeks of diabetes induced significant vascular dysfunction, as assessed by abnormally high vasoconstrictor responses to norepinephrine in the mesenteric vascular bed (MVB). Diabetes also led to increased phosphorylation of all ErbB receptors with subsequent increased phosphorylation of downstream ERK1/2 in the MVB. Chronic 4-week in vivo administration of a novel pan-ErbB inhibitor, generation 7 polyamidoamine dendrimer (G7PAMAM; 20mg/kg daily i.p), or the well-established pan-ErbB inhibitor, Genistein (1mg/kg i.p. alt diem) to diabetic rats led to a marked attenuation in the hyperphosphorylation of all ErbBs, ERK1/2 and to a significant correction in NE-mediated hyper-responsiveness of the diabetic MVB. Furthermore, pan-ErbB inhibitors attenuated high glucose-induced ErbBs-ERK1/2 signaling in primary vascular smooth muscle cells grown in high (25mM) glucose. These data show for the first time that pan-ErbB inhibition attenuates diabetes-induced vascular dysfunction in an experimental model of diabetes. Therefore, pan-ErbB inhibition might represent a novel strategy for the treatment of diabetes-induced vascular complications.

Disclosure
S. Akhtar: None. B. Chandrasekhar: None. I. Benter: None.

Funding
Qatar University (QUCG-CMED-19/20-3)
463-P: Deterioration of Atherosclerosis via Dysbiosis in ApoE Null Mice Fed with High-Salt Diet
Authors: KIMURA TOMONORI , HASHIMOTO YOSHITAKA , SENMARU TAKAFUMI , USHIGOME EMI , HAMAGUCHI MASAHIDE , YAMAZAKI MASAHIRO , FUKUI MICHIAKI 
Published: June 2020

Recent studies reported that the impairment of gut microbiota (Dysbiosis) cause impairments of gut metabolisms, which induce chronic inflammation in the body. High salt intake not only increases blood pressure, but also causes Dysbiosis. However, the detailed mechanism has not been clarified yet. Here we investigate the mechanism of the atherosclerosis progression through dysbiosis caused by high salt diet. 16-week-old male Apolipoprotein E-deficient (ApoE-/-) mice were fed normal breeding diet (ND group), high-fat high-sucrose diet (HFHSD group), and high-fat high-sucrose high-salt diet (HFHSDNaCl8%, high salt group) for 8 weeks. The concentration of salt in ND and HFHSD is set as 2%. Immune cells in the aorta were extracted and evaluated by FACS CANTII. The metagenome was evaluated by the 16S target sequencing method. Lipidome in the aorta, serum, or intestinal contents were evaluated and quantified by GCMS system (Agilent 7890B/5977B). In high salt group, gene expression levels of inflammatory cytokines and chemokines were upregulated, and M1 macrophages and Group 2 innate lymphoid cells in atherosclerotic lesions increased. The prevalence of Firmicutes decreased and the prevalence of Bacteroidetes increased. Lipidome in the aorta, serum and intestinal contents were significantly different among the HFHSD group and high salt group, although the lipids administered were the same. High salt intake increases M1 macrophages and Group 2 innate lymphoid cells by altering intestinal bacterial lipid metabolisms, and could deteriorate chronic inflammation of arteriosclerosis.

Disclosure
T. Kimura: None. Y. Hashimoto: None. T. Senmaru: None. E. Ushigome: None. M. Hamaguchi: None. M. Yamazaki: None. M. Fukui: None.
465-P: Thrombin Induces SGLT1 and SGLT2 Expression to Promote the AT1R/NADPH Oxidase-Mediated Pro-oxidant Response Inducing Senescence in Atrial Endothelial Cells
Authors: HASAN HIRA , PARK SIN-HEE , BELCASTRO EUGENIA , AUGER CYRIL , LEE HYUN-HO , SCHINI-KERTH VALERIE 
Published: June 2020

Selective sodium-glucose cotransporter 2 (SGLT2) inhibitors have shown cardiovascular protection in type 2 diabetes patients with established cardiovascular (CV) disease independently of glycemic control. Although blood-derived coagulation factors are major contributors of CV diseases, their impact on the expression of SGLT1 and 2 in endothelial cells (ECs) has not been studied. This study investigated whether thrombin affects the expression of SGLT1 and 2 and examined their role in the induction of endothelial senescence.
ECs were isolated from porcine atrial tissue (AECs) and used at first passage. Endothelial senescence was assessed by determining senescence-associated beta-galactosidase (SA-beta-gal) activity, oxidative stress using dihydroethidium, and protein level by Western blot analysis and immunofluorescence staining.
Exposure of AECs to thrombin (1 or 3 U/ml) for 24 h induced an upregulation of the low protein expression level of SGLT1 and SGLT2. Thrombin induced a sustained pro-oxidant response and increased the level of SA-beta-gal activity that were both inhibited by antioxidants, an angiotensin-converting enzyme inhibitor and an AT1 receptor antagonist and also by sotagliflozin (a dual SGLT1 and SGLT2 inhibitor) and by empagliflozin (a selective SGLT2 inhibitor). In addition, thrombin upregulated the expression level of angiotensin-converting enzyme and AT1 receptors, VCAM-1 and down-regulated that of endothelial NO synthase.
In conclusion, thrombin up-regulates both SGLT1 and SGLT2 expression in AECs to sustain oxidative stress leading to endothelial senescence and dysfunction. The fact that the AT1R/NADPH oxidase/SGLT1 and 2 pathway acts in a feedforward manner suggests that inhibition of SGLT1 and/or SGLT2 appears as an attractive strategy to perpetuate the protective endothelial function on the cardiovascular system.

Disclosure
H. Hasan: None. S. Park: None. E. Belcastro: None. C. Auger: None. H. Lee: None. V. Schini-Kerth: Research Support; Self; Boehringer Ingelheim International GmbH.
47-OR: A Reference Map of the Human Proinsulin Biosynthetic Interaction Network
Authors: TRAN DUC T. , ARVAN PETER , KAUFMAN RANDAL J. , ITKIN-ANSARI PAMELA 
Published: June 2020

The beta-cell protein synthetic machinery is dedicated to insulin production, and recent reports suggest misfolding of its precursor, proinsulin, is a feature of type 2 diabetes (T2D). Despite the critical role of insulin in organismal homeostasis, the precise network of interactions required for proper proinsulin folding and subsequent trafficking through the Golgi and secretory pathway, remains poorly defined. In the present study we conducted unbiased profiling of the proinsulin interactome in human islets, utilizing a human proinsulin-specific monoclonal antibody for affinity purification and mass spectrometry. Stringent analysis identified a central node of interactions between human proinsulin and sequential secretory pathway proteins that is remarkably conserved across both genders and 3 ethnicities. Among the most prominent proinsulin interactions was with the ER localized oxidoreductase peroxiredoxin-4 (PRDX4). A functional role for PRDX4 was demonstrated by knockdown studies in MIN6 beta-cells that rendered proinsulin susceptible to misfolding, particularly in response to oxidative stress. Conversely, exogenous PRDX4 improved proinsulin folding. Notably, oxidative stress and even high glucose treatment alone induced proinsulin misfolding in human islets and MIN6 cells and this was accompanied by sulfonylation of PRDX4, a modification known to inactivate peroxiredoxins. This finding prompted PRDX4 analysis in a panel of human islet samples that revealed significantly higher levels of sulfonylated (inactive) PRDX4 in islets from patients with T2D compared to that of healthy individuals. Taken together, these data provide a detailed reference map of the human proinsulin interaction network and suggest critical unrecognized areas for study in insulin biosynthesis, beta cell function, and T2D.

Disclosure
D.T. Tran: None. P. Arvan: None. R.J. Kaufman: None. P. Itkin-Ansari: None.

Funding
National Institutes of Health (R24DK110973); JDRF (2-SRA-2015-47-M-R)
480-P: Reversal of Diabetic Nephropathy in T2D db/db Mouse Model by Systemic Pyruvate Kinase M2 (PKM2) Activation
Authors: ST-LOUIS RONALD , FU JIALIN , PARK KYOUNGMIN , SHINJO TAKANORI , LUDEKE JOHANNA , YOKOMIZO HISASHI , LI QIAN , WOLFSON EMILY , KING GEORGE L. 
Published: June 2020

Mitochondrial abnormalities induced by diabetes have been postulated to cause several complications including nephropathy. Elevated expressions of enzymes for glycolysis and mitochondrial functions in the glomeruli have been associated with protection against the development of diabetic nephropathy of chronic duration. Activation of a glycolytic enzyme, pyruvate kinase M2 (PKM2), by a small molecule selective activator (TEPP-46) or its targeted overexpression to the podocytes in mice reversed glomerular mitochondrial dysfunction and pathology in insulin deficient diabetic mice. However, the effects of PKM2 activation to stop glomerular dysfunction and pathology have not been studied in animal models of type 2 diabetes (T2D). Thus, we studied the effect of activating glycolysis and mitochondrial metabolism with PKM2 activator, TEPP-46, in db/db mice (with obesity and hyperglycemia) and BLKS mice on regular diet (RD) or high fat diet (HFD). HFD feeding induced obesity in BLKS but did not cause changes in glomerular PKM activities or functions as measured by urinary albumin to creatinine ratio (ACR). Oral feeding with TEPP-46 did not affect body weight (BW), glucose tolerance, lipids, ACR, glomerular pathology in BLKS mice on RD or HFD. In contrast, db/db mice on RD had elevated BW and hyperglycemia that were significantly increased by HFD, which also induced loss of glomerular PKM2 protein expression and activities, elevations of gene expression for fibrosis (fibronectin, TGF-b), mesangial expansion and ACR after 3 months. Intervention with oral TEPP-46 treatment for 3 months, but not 1 month, after 3 months of diabetes and HFD, (total of 6 months of diabetes), improved significantly both glomerular PK activities and ACR, without lowering hyperglycemia and hyperlipidemia of the db/db mice. These findings support the idea that systemic activation of PKM2 can improve renal functions in both T1D and T2D.

Disclosure
R. St-Louis: None. J. Fu: None. K. Park: None. T. Shinjo: None. J. Ludeke: None. H. Yokomizo: None. Q. Li: None. E. Wolfson: None. G.L. King: Research Support; Self; Janssen Pharmaceuticals, Inc.
489-P: Energy Status Could Modulate Autophagy through Colocalizations of Fyn, P53, and Tgm2 in the Renal Tubules In Vivo
Authors: UEHARA RYOTA , YAMADA EIJIRO , NAKAJIMA YASUYO , HORIGUCHI KAZUHIKO , ISHIDA EMI , MATSUMOTO SHUNICHI , OKADA SHUICHI , BASTIE CLAIRE C. , YAMADA MASANOBU 
Published: June 2020

Diabetic kidney disease (DKD) is one of the major diabetic complications, the leading cause of the end stage renal disease, and recently autophagy was shown to regulate DKD. Previously we reported Fyn functions as a muscle mass regulator by suppressing autophagy. More recently, we demonstrated that Fyn also down-regulates autophagy in HK2 cells, an in vitro cell model of renal proximal tubular epithelial cells (RPTC). Phospho-proteomic analysis revealed that Fyn phosphorylates Transglutaminase 2 (Tgm2), a known autophagic inhibitor, on Y369 and Y617. Fyn dependent phosphorylation of Tgm2 regulates autophagy. Moreover, since Tgm2 was reported to make a complex with p53 and p62, known autophagy regulator/substrate to degrade p53 in autophagosome in cancer cells we examined it in HK2 cells and found that p53 expression was decreased in Tgm2 knock-downed HK2 cells indicating that Tgm2-p62-p53 complex modulating autophagy in RPTC. Those are biochemical studies with in vitro systems, we proceeded it in vivo. Since previously we showed that Fyn could be regulated by energy status, we first examined whether energy status could change the subcellular localization of those molecules in RPTC in vivo using the marker, Aquaporin 1. Confocal microscopic studies revealed that Ad libitum fed mice showed no punctate of p62 in those cells indicating suppressing autophagy. Fyn, Tgm2 and p53 shaped the dotted form mainly in basement membrane. Interestingly, all the molecules changed their localizations into cytoplasm in fasted state where p62 punctations observed indicating increased autophagy. Taken together, these data suggest that Energy status could regulates Fyn to not only phosphorylates Tgm2 and modulates Tgm2-p62-p53 complex but also change their co-localizations of Fyn, p53 and Tgm2 in RPTC to regulate autophagy leading to pathogenesis of DKD.

Disclosure
R. Uehara: None. E. Yamada: None. Y. Nakajima: None. K. Horiguchi: None. E. Ishida: None. S. Matsumoto: None. S. Okada: None. C.C. Bastie: None. M. Yamada: None.