Normoglycemic Research Articles

Prediabetes, diabetes, metabolic syndrome, and small fiber neuropathy.

This study was conducted to evaluate the association between prediabetes (PD), elements of the metabolic syndrome (MetS), and small fiber neuropathy (SFN). A total of 268 patients with SFN symptoms and normal electrophysiology underwent tests to assess small fibers. SFN was diagnosed based on abnormality of at least two among intraepidermal nerve fiber density (IEFND), quantitative sensory testing, and quantitative sudomotor axon reflex testing. There was no difference in IENFD or abnormal skin biopsy frequency between PD and normoglycemia (NG). However, IENFD was lower in people with diabetes mellitus (DM) than in those with NG. An association between HbA1C and IENFD was observed only if DM patients were included. Attributes of the MetS were more common in those with an abnormal skin biopsy but not among those with autonomic dysfunction or meeting SFN criteria. DM, but not PD alone, is associated with SFN. Other MetS elements appear to preferentially impact small fiber structure over function.

Negative impact of gestational diabetes mellitus on progress of pelvic floor muscle electromyography activity: Cohort study.

Background and objectivePelvic floor muscles are involved in postural stability, in maintenance intra-abdominal pressure, and on mechanical support for pelvic organ. Gestational Diabetes Mellitus’ (GDM) pregnancies complicated by fetal macrosomia, large placenta and polyhydramnios contribute for abrupt and intense increase in maternal intra-abdominal pressure. Our objective was analyze the impact of GDM on pelvic floor muscle (PFM) electromyography (EMG) activity progress from 24–30 to 36–38 weeks of gestation. We conducted a prospective cohort study. PFM EMG was performed in nulliparous or primiparous women with one previous elective cesarean delivery and with or not GDM diagnosed by the American Diabetes Association criteria. A careful explanation of the muscle anatomy and functionality of the PFM was given before EMG assessment. The outcome measures were PFM recruitment and progress from 24–30 to 36–38 weeks of gestation analyzed by the normalized root mean square (RMS) during rest-activity, fast and hold pelvic floor muscle contraction.ResultsFifty-two pregnant women were assigned to 2 groups: the GDM (n = 26) and normoglycemic (NG) (n = 26). The demographic and obstetric data showed homogeneity between the groups. PFM activity progress was decreased in rest-activity (P = 0.042) and hold contraction (P = 0.044) at 36–38 weeks of gestation in the GDM group relative to that in the NG group.ConclusionGDM group showed a progressive decrease in EMG-PFM activity during rest-activity and hold contractions from 24–30 to 36–38 weeks of gestation.

Calcifediol Decreases Interleukin-6 Secretion by Cultured Human Trophoblasts From GDM Pregnancies.

Gestational diabetes mellitus (GDM) is often characterized by low maternal calcifediol (25OHD) and high inflammation levels. This study aimed to determine whether placental protein expressions of CYP27B1, vitamin D receptor (VDR), and CYP24A1 are impaired in GDM and to investigate the effect of a 25OHD treatment on IL-6 secretion by GDM trophoblasts compared with normoglycemic (NG) trophoblasts. Placental tissue samples were harvested to determine protein expression of CYP27B1, VDR, and CYP24A1 by immunoblots. Isolated trophoblasts were stimulated with 25OHD concentrations (25 to 2000 nM) once a day for 3 days and IL-6 secretion was quantified (ELISA). We recruited 17 NG women, 19 women with GDM treated with diet and exercise alone (GDM-d) and 9 women with GDM who necessitated insulin therapy (GDM-i). Protein expressions of CYP27B1 and VDR were significantly higher in placental tissue from GDM-d women compared with NG women (both P = 0.02), whereas no differences were detected between GDM-i and NG placental tissues. In cultured trophoblasts (two groups; n = 5 NG and n = 5 GDM-d), exposure to increasing 25OHD concentrations significantly decreased IL-6 secretion in the GDM-d group only (P = 0.006). After treatment with 25OHD (2000 nM), IL-6 secretion was lower in the GDM-d group compared with the NG group (P = 0.03). Our results suggest an upregulation of the VDR-1,25(OH)2D complex bioavailability in GDM-d placentas, possibly reflecting a compensatory mechanism aiming to ensure that vitamin D can exert its genomic and nongenomic effects in the target cells of the placental-fetal unit. Our findings support an anti-inflammatory effect of vitamin D at the feto-maternal interface in GDM-d pregnancies.

509-P: Utility of Serum Cystatin C in Early Kidney Dysfunction in Prediabetics

Background: Serum cystatin C (sCys C) has been proposed as a marker of kidney function with advantages compared to creatinine. It is unclear whether renal damage begin in prediabetes (PDM), when sCys C could be a promisor marker of early kidney dysfunction. We investigated whether sCys C differed between normoglycemic (NG) and PDM subjects and contributed to the detection of kidney dysfunction. Methods: A cross-sectional analysis of 947 nondiabetic participants (mean age 45.7 years) from the Brazilian Longitudinal Study of Adult Health was performed. sCys C (determined by ELISA) and CKD-EPI eGFR were compared between NG and PDM (fasting plasma glucose: 100-125 mg/dL or 2-hour post-load: 140-199 mg/dL or A1c: 5.7-6.4%). PDM were stratified in 4 groups according to altered sCys C and albumin excretion rate (AER): normal sCys C and AER (G1), abnormal sCys C and normal AER (G2), normal sCys C and abnormal AER (G3) and abnormal sCys C and AER (G4) were compared using ANOVA. Results: PDM subjects (n = 671) had anthropometric and biochemical data higher than NG (n=276), higher sCys C levels [0.67 (0.41-0.95) vs. 0.48 (0.31-0.81), p<0.001] and lower eGFR (96.3±17.4 vs. 100.6±17.1 mL/min/1.73m2, p<0.001). In eGFR categories (60-<90, 90-<125 and ≥125 mL/min/1.73m2), sCys C were always higher in PDM subjects than in NG. Comparing NG individuals, hyperfiltrants had lower sCys C than normofiltrants (p=0.03) but not than mildly reduced eGFR individuals (p=0.12). Considering the PDM groups, eGFR gradually dropped from G1 to G4 (96.8±17.4 vs. 96.2±16.9 vs. 94.0±17.2 vs. 77.2±25.4 mL/min/1.73m2, p-trend = 0.06) and, as expected, mean eGFR in G4 was lower than in G1 (p=0.017). Conclusion: Our data confirmed that sCys C is increased in PDM. Reduced sCys C only in NG hyperfiltrant subjects could suggest an inability of hyperfiltrant PDM to excrete Cys C. No statistical evidence that sCys C could be an early marker of kidney dysfunction in prediabetes was found; prospective studies should address its utility to detect dysfunction before AER elevation. Disclosure J.I.F. Branda: None. B. Almeida: None. S.R.G. Vivolo: None.

403-P: Cerebral Blood Flow Responses to Hypoglycemia in Healthy Humans Treated with Induction Protocol to Produce Impaired Awareness of Hypoglycemia

In this study, we sought to determine by 3T MRI the cerebral blood flow (CBF) responses to hypoglycemia (HG) in nondiabetic controls exposed to either two euglycemic or two hypoglycemic clamps on the day before the MRI (EU- or HYPO-preconditioning, respectively). Under the belief that HYPO-preconditioning induces hypoglycemia-associated autonomic failure (HAAF) with blunting of the counter-regulatory hormone and symptom responses to HG (Arbelaez et al, 2008), we hypothesized that CBF responses after HYPO-preconditioning would resemble those reported in T1D subjects with impaired awareness of hypoglycemia (IAH) (Weigers et al, 2017). CBF was measured by arterial spin labeling during a 2-step hyperinsulinemic clamp [step 1 = 95 mg/dL normoglycemia (NG), step 2 = 50 mg/dL (HG)]. Symptom and counter-regulatory hormone were also measured. Full CBF datasets were obtained from 10 participants who underwent EU-preconditioning (7F/3M, age 37 ± 14 years, BMI 27.2 ± 3.7 kg/m2) and 9 who underwent HYPO-preconditioning (5F/4M, age 40 ± 14 years, BMI 26.2 ± 2.6 kg/m2). In contrast with our assumption, the HYPO-preconditioning did not induce blunting of symptom or counter-regulatory hormone responses to HG other than for glucagon (33 ± 19 and 14 ± 21 pg/mL for EU- and HYPO-preconditioning, respectively, p=0.04 uncorr). CBF responses did not differ between the two pre-conditionings, and resembled those reported in the literature in naïve nondiabetic controls (Weigers et al, 2017). In particular, the cortical grey matter CBF did not change during HG vs. NG for either the EU- or HYPO-preconditioning settings (1 ± 10% and 2 ± 10%, respectively), while thalamic CBF increased in both settings (21 ± 20%, p=0.012 corr, and 27 ± 30%, p=0.06 corr, respectively). We conclude that the preconditioning protocols should be used with caution in nondiabetic controls for investigating IAH, as the mechanisms of HAAF in T1D patients may be specific to their disease condition. Disclosure A. Canna: None. H. Grohn: None. D. Mascali: None. P. Filip: None. A. Moheet: None. A. Kumar: None. X. Li: None. E.A. Olawsky: None. L.E. Eberly: None. F. Esposito: None. E.R. Seaquist: Advisory Panel; Self; Eli Lilly and Company, Zucara Therapeutics Inc. Consultant; Self; 360 Consulting, MannKind Corporation. Research Support; Self; Eli Lilly and Company, JDRF, National Institutes of Health. Other Relationship; Self; American Diabetes Association, Sanofi, WebMD. S. Mangia: None. Funding National Institutes of Health

Antihyperglycemic activity of the leaves from annona diversifolia Safford. and farnesol on normal and alloxan-induced diabetic mice*

Background: Diabetes mellitus (DM) is a chronic disease characterized by high blood glucose levels resulting from insulin resistance or inadequate insulin secretion. In the world, DM is one of the most frequent non-contagious diseases that affect more than 371 million people. Objective: This study aimed to evaluate the antihyperglycemic properties of the ethanol extract, subsequent fractions, and farnesol obtained from the leaves of Annona diversifolia on alloxan-induced diabetic and normal mice. Materials and Methods: Bioassay-guided fractionation of the ethanol extract of the leaves of A. diversifolia (EELAd) was performed on alloxan-induced Type 2 diabetic and normoglycemic (NM) mice. Oral glucose tolerance test (OGTT), oral sucrose tolerance test (OSTT), and oral lactose tolerance test (OLTT) were performed in fast NM mice (FNM). Results: The EELAd, CHCl3 fraction, and farnesol induced a significant reduction of postprandial hyperglycemia in acute and subchronic tests using AITD mice. When EELAd, CHCl3 fraction, and farnesol were tested on NM in subchronic assays, these did not affect glycemic levels. In the case of acute test on NM, only CHCl3 fraction induced a hypoglycemic effect at 2 h after the treatment. OLTT and OSTT showed that the EELAd, CHCl3 fraction, and farnesol induced a significant reduction of hyperglycemia levels in FNM at 2 h after a lactose or sucrose load comparable to acarbose. In the case of OGTT was observed a significant reduction of hyperglycemia levels in FNM mice at 2 h after a glucose load comparable to canagliflozin. Conclusion: The EELAd and farnesol induced a significant reduction of postprandial hyperglycemia on AITD mice in acute and subchronic assays. Our results suggest that the control of postprandial hyperglycemia may be mediated by the regulation of absorption of glucose and inhibition of disaccharide digestion such as sucrose and lactose. Finally, the results explained the use of A. diversifolia in Mexican traditional medicine as an antihyperglycemic agent.

Effects of Metformin Therapy on Coronary Endothelial Dysfunction in Patients With Prediabetes With Stable Angina and Nonobstructive Coronary Artery Stenosis: The CODYCE Multicenter Prospective Study.

To evaluate the effect of metformin therapy on coronary endothelial function and major adverse cardiac events (MACE) in patients with prediabetes with stable angina and nonobstructive coronary stenosis (NOCS). Metformin therapy may be needed to reduce coronary heart disease risk in patients with prediabetes. A total of 258 propensity score-matched (PSM) patients with stable angina undergoing coronary angiography were enrolled in the study. Data from 86 PSM subjects with normoglycemia (NG), 86 PSM subjects with prediabetes (pre-DM), and 86 PSM subjects with prediabetes treated with metformin (pre-DM metformin) were analyzed. During coronary angiography, NOCS was categorized by luminal stenosis <40% and fractional flow reserve >0.80. In addition, we assessed the endothelial function, measuring coronary artery diameter of left anterior descending coronary (LAD) at baseline and after the infusion of acetylcholine, by means of an intracoronary Doppler guide wire. MACE, as cardiac death, myocardial infarction, and heart failure, was evaluated at 24 months of follow-up. At baseline, NG patients had a lower percentage of LAD endothelial dysfunction compared with pre-DM patients (P < 0.05). The pre-DM patients had a higher percentage of endothelial LAD dysfunction as compared with the pre-DM metformin patients (P < 0.05). At the 24th month of follow-up, MACE was higher in pre-DM versus NG (P < 0.05). In pre-DM metformin patients, MACE was lower compared with pre-DM patients (P < 0.05). Metformin therapy may reduce the high risk of cardiovascular events in pre-DM patients by reducing coronary endothelial dysfunction.

IL-10 Could Play a Role in the Interrelation between Diabetes Mellitus and Osteoarthritis.

The association between osteoarthritis (OA), obesity and metabolic syndrome suggests an interrelation between OA and diabetes mellitus (DM). Little is known about the role of anti-inflammatory cytokine interleukin (IL)-10 in the interrelation between OA and DM. Hence, the effects of IL-10 under hyperglycemia (HG) and hyperinsulinemia (HI) in human articular chondrocytes (hAC) and chondrosarcoma cell line Okayama University Medical School (OUMS)-27 were examined. HAC and OUMS-27, cultured in normoglycemic (NG) and HG conditions were stimulated with insulin and/or IL-10. Cell survival, metabolic activity, proliferation and extracellular matrix (ECM) synthesis were immunocytochemically examined. No significant differences in vitality of hAC neither in pure NG (NGw/o) nor HG (HGw/o) conditions were found. Applying HI and/or IL-10 in both conditions reduced significantly the vitality of hAC but not of OUMS-27. HG impaired significantly hAC metabolism. When combined with HI + IL-10 or IL-10 alone it decreased also significantly hAC proliferation compared to NGw/o. In OUMS-27 it induced only a trend of impaired proliferation compared to NGw/o. hAC but not OUMS-27 reduced significantly their collagen type (col) I, SOX9 and proteoglycan (PG) synthesis in HG combined with HI +/− IL-10 compared to NGw/o. IL-10 could not moderate HI and HG effects. In contrast to hAC OUMS-27 showed limited sensitivity as DM model.

ANTIHYPERGLYCEMIC, ANTIOXIDANT AND ANTI-INFLAMMATORY EFFECTS OF AQUEOUS EXTRACT OF MISTLETOE (Cladocolea loniceroides) IN STZ-INDUCED DIABETIC MICE

Background: Inhibition of carbohydrate hydrolyzing enzymes, such as α-amylase and α-glucosidase, is a key element in the regulation of diabetes mellitus (DM). The purpose of this work was to study the inhibition of carbohydrate hydrolyzing enzymes, and the antihyperglycemic activity of aqueous extract of Cladocolea (C. loniceroides) in streptozotocin (STZ)- induced diabetic mice. Materials and Methods: The inhibitory activities of C. loniceroides aqueous extract on α-amylase and α-glucosidase were investigated in vitro. Glucose tolerance test was performed in normoglycemic (NG) mice which were fed with starch or sucrose. The effect of mistletoe aqueous extract (ME) was measured in (STZ)-induced diabetic mice. On day 35 of the treatment, the effect of decreasing oxidative stress (lipid peroxidation, glutathione redox state, GPx and GR specific activities, cytokines and aminotransferases analysis) was assessed. Results: ME showed a competitive mode of inhibition for the carbohydrate hydrolyzing enzymes (CHE). The maximum antihyperglycemic activity in mice was observed for the unripe fruit aqueous extract (UFAE) for -amylase and stem aqueous extract (SAE); for -glucosidase due to the glycemic response reduction by 23% or 35%, respectively. UFAE decreased malondialdehyde (MDA) 1.76 times; GSH/GSSG ratio was mantained (3.08 ± 0.66); GPx activity was reduced (24%); IL-6 was reduced (18%) and the concentration of TNF- (37%) was leveled with respect to the (STZ)-induced diabetic mice; ALT and AST (liver transaminases) levels were nearly the same compared with those found in the NG mice. Conclusion: UFAE of C. loniceroides exhibited the highest antidiabetic activity in (STZ)-induced diabetic mice.

Hospitalized patients with stress hyperglycemia: Incidence of diabetes and mortality on follow-up

IntroductionThe study objective was to estimate during post-discharge follow-up the incidence of diabetes and to ascertain mortality in hospitalized patients, classified during follow-up as having stress hyperglycemia (SH) or normoglycemia (NG) based on blood glucose levels. Material and methodsA retrospective cohort of non-diabetic adults with SH (>140mg/dl and HbA1c<6.5%) or NG (all blood glucose values ≤140mg/dl) was used. ResultsThere were 3981 patients with NG and 884 with SH. During the observation period (median follow-up of 1.83 years), there were 255 cases of diabetes and 831 deaths. The cumulative incidence of diabetes per year was 1.59% (95%CI: 1.23–2.06) in patients with NG and 7.39% (95%CI: 5.70–9.56) in those with SH. SH was significantly associated to diabetes (crude HR 1.33, 95%CI: 1.13–1.73, p 0.025), even after adjusting for age and sex (adjusted HR 1.38, 95%CI 1.06–1.78, p 0.014). The mortality rate at one year was 10.07% (95%CI: 9.18–11.05) in NG patients and 13.24% (95%CI: 11.17–15.65) in SH patients. The sub-hazard ratio of developing diabetes considering death as a competitive event was 1.41 (95%CI 1.29–1.53, p<0.001). ConclusionsSH is a risk factor for diabetes. There were no differences in mortality during follow-up, but death appears to be a competitive event in development of diabetes in this population.

Involvement of Beclin‑1 in axonal protection by short‑term hyperglycemia against TNF‑induced optic nerve damage

Beclin-1 serves a pivotal role in autophagosome formation. A previous study demonstrated that streptozotocin-induced hyperglycemia (HG) ameliorates axonal loss induced by tumor necrosis factor (TNF) with upregulation of autophagy in rats. The aim of present study was to examine whether Beclin-1 is involved in this autophagy machinery. Immunoblot analysis of optic nerves demonstrated that HG upregulated Beclin-1 protein expression when compared with normoglycemia (NG). Intravitreal administration of TNF did not alter the optic nerve Beclin-1 expression in NG nor in HG. Beclin-1 immunoreactivity was revealed to be mainly in astrocytes in optic nerves; however, it was also observed in the neurofilaments of the HG group. Morphometric analysis revealed that HG appeared to have substantial ameliorative effects on axon loss and this ameliorative effect was partially prevented by Beclin-1 small interfering RNA. These results indicated that Beclin-1 may exist in neurons and glia in optic nerves and increased Beclin-1 expression may be at least partially associated with axonal protection by HG.

Pacientes hospitalizados con hiperglucemia de estrés: incidencia de diabetes y mortalidad al seguimiento

IntroductionThe study objective was to estimate during post-discharge follow-up the incidence of diabetes and to ascertain mortality in hospitalized patients, classified during follow-up as having stress hyperglycemia (SH) or normoglycemia (NG) based on blood glucose levels. Material and methodsA retrospective cohort of non-diabetic adults with SH (> 140mg/dl and HbA1c <6.5%) or NG (all blood glucose values ≤ 140mg/dl) was used. ResultsThere were 3981 patients with NG and 884 with SH. During the observation period (median follow-up of 1.83 years), there were 255 cases of diabetes and 831 deaths. The cumulative incidence of diabetes per year was 1.59% (95% CI: 1.23-2.06) in patients with NG and 7.39% (95% CI: 5.70-9.56) in those with SH. SH was significantly associated to diabetes (crude HR 1.33, 95% CI: 1.13-1.73, p .025), even after adjusting for age and sex (adjusted HR 1.38, 95% CI 1.06-1.78, p .014). The mortality rate at one year was 10.07% (95% CI: 9.18-11.05) in NG patients and 13.24% (95% CI: 11.17-15.65) in SH patients. The sub-hazard ratio of developing diabetes considering death as a competitive event was 1.41 (95% CI 1.29-1.53, p <.001). ConclusionsSH is a risk factor for diabetes. There were no differences in mortality during follow-up, but death appears to be a competitive event in development of diabetes in this population.

1-Hour and 2-Hour Postload Glucose Level on Oral Glucose Tolerance Test and the Risk of Incident Metabolic Syndrome.

Metabolic syndrome (MetS) increases the cardiometabolic risk even in nondiabetic patients. Previous studies have demonstrated that 1-hour postload glucose (PG) and 2-hour PG based on oral glucose tolerance test (OGTT) predicted cardiometabolic risk. However, it is still unclear whether and to what extent postload glucose is associated with the risk of MetS. A total of 5389 nondiabetic Koreans were dichotomized into normoglycemic (NG) groups and abnormal glycemic groups based on OGTT, including elevated 1-hour PG (155 to 199 mg/dL) and impaired glucose tolerance (IGT) (2-hour PG 140 to 199 mg/dL), and followed up for 10 years. Cox proportional hazard model was used to evaluate hazard ratios (HRs) with 95% CIs for incident MetS. Subgroups were determined by high or normal 1-hour PG (cutoff: 155 mg/dL) and 2-hour PG (cutoff: 140 mg/dL). Compared with NG, the risk of MetS increased proportionally to the level of 1-hour PG and 2-hour PG, independently of the number of baseline metabolic components. Even within people with normoglycemia, elevated PG above specific levels (1-hour PG ≥115 mg/dL and 2-hour PG ≥100 mg/dL) was significantly associated with the increased risk of MetS. In subgroup analysis, adjusted HR for MetS was higher in the group with high 1-hour PG and normal 2-hour PG [1.53 (95% CI, 1.35 to 1.74)] than in the group with normal 1-hour PG and high 2-hour PG [1.32 (95% CI, 1.02 to 1.70)]. Elevated 1-hour PG and 2-hour PG significantly are associated with greater risk for MetS, and 1-hour PG was superior to 2-hour PG in predicting MetS.

TCF7L2 gene variants predispose to the development of type 2 diabetes mellitus among individuals with metabolic syndrome.

Transcription factor 7-like 2 (TCF7L2) gene variants rs12255372 and rs7903146 have been consistently shown to raise genetic risk for type 2 diabetes mellitus (T2DM). The aim of this study was to investigate the possible role of these variants in the development of impaired glucose metabolism (IGM), including impaired fasting glucose (IFG) or T2DM, in patients with metabolic syndrome (MS). The study population consisted of 228 patients with MS who were divided into two groups. The first group consisted of 148 patients with MS and IGM [39M/109F, 59.8 ± 14.6 (mean ± SD) years] and the second group of 80 patients with MS and normoglycemia (NGM) (16M/64F, 56.1 ± 15.8years). The diagnosis of MS was based on the criteria proposed by the American Heart Association/National Heart, Lung, and Blood Institute (AHA/NHLBI) Scientific Statement. Anthropometric parameters including BMI and waist circumference were recorded and blood samples were obtained after overnight fasting for biochemical tests. The rs12255372 and rs7903146 TCF7L2 polymorphisms were genotyped in peripheral blood leucocytes. Analysis of the distribution of the TCF7L2 polymorphic alleles revealed that the frequency of the T allele of the TCF7L2 variant rs12255372 was 38.2% in the study population, while the frequency of the T allele of the TCF7L2 rs7903146 variant was 35.3%. The T allele of the rs12255372 variant was more frequently present in patients with MS and IGM (48.3%) compared to patients with MS and NGM (19.4%, p < 0.001). Also, the T allele of rs7903146 was more frequently present in patients with MS and IGM (44.6%) compared to patients with MS and NGM (18.1%, p < 0.001). Logistic regression analysis followed and revealed that the presence of the T allele for both rs12255372 and rs7903146 TCF7L2 gene variants is a very powerful predictor of the presence of glucose disorders, increasing the risk more than fourfold in patients with MS and after adjustment for potential confounders, such as age, gender, BMI, and waist circumference (TCF7L2 rs12255372: Exp(B) 4.917, p < 0.001 and TCF7L2 rs7903146: Exp(B) 5.460, p < 0.001). The presence of the rs12255372 and rs7903146 TCF7L2 gene variants plays an important role in the development of T2DM among individuals with MS. These findings support the notion that among subjects with MS, those who finally develop T2DM have a genetic predisposition to β-cell dysfunction.

The antioxidant gallic acid induces anxiolytic-, but not antidepressant-like effect, in streptozotocin-induced diabetes.

The physiopathology of anxiety or depression related to diabetes is still poorly understood. The treatment with antidepressant drugs is a huge challenge due to theirs adherence low rate and many adverse effects. Thus, the seeking for a better treatment for these associated diseases is of utmost importance. Given that the oxidative stress in different tissues occurs in diabetes and anxiety or depression as well, the antioxidant gallic acid becomes an interesting compound to be investigated. Thus, the effects of long-term treatment with gallic acid (0, 10, 20 and 40mg/kg; gavage) were evaluated in diabetic (DBT) animals submitted to the elevated plus-maze (EPM), the light-dark transition (LDT) tests and modified forced swim test (mFST). Also, indirect parameters of oxidative stress, lipid peroxidation (LPO) and reduced glutathione (GSH) levels were evaluated in the hippocampus (HIP) and prefrontal cortex (PFC). The results showed that DBT animals presented a decrease in the spent time in the open arms, in the end arm exploration and head dips when evaluated in the EPM test; moreover, a decrease in the spent time in the lit compartment of LDT test was observed, suggesting an anxiogenic-like behavior. During the mFST, an increase in the mean counts of immobility and a decrease in the mean counts of swimming and climbing were observed, indicating a depressive-like behavior. These aversive behaviors were more pronounced when compared to normoglycemic (NGL) animals and streptozotocin-treated animals that not become DBT. In addition, DBT rats showed an increase in the oxidative stress parameters in the HIP and PFC that was reversed by the gallic acid treatment (lowest dose - 10mg/kg), i.e., the treatment decreased the elevated LPO levels and increased the reduced GSH in the HIP and PFC. Also, gallic acid treatment was able to produce an anxiolytic-like effect in the EPM and LDT tests, but not antidepressant-like effect in the FST. Taken together, the results suggest that the antioxidant/neuroprotective effect of gallic acid treatment in HIP and PFC of DBT animals may be essential to the anxiolytic-like effect.

LIK066, a Dual SGLT1/2 Inhibitor, Reduces Weight and Improves Multiple Incretin Hormones in Clinical Proof-of-Concept Studies in Obese Patients With or Without Diabetes

Background: LIK066 is a potent dual inhibitor of SGLT1/2 (IC50: 20.6 and 0.58 nM for SGLT1 and SGLT2). We studied mechanistic effects of dual SGLT1/2 inhibition of SGLT1 in the gut and SGLT1/2 in the kidneys. Methods: Effects of LIK066 on weight, glucose and a variety of incretin hormones (GLP-1, PYY, GIP, glucagon, insulin) were investigated in: 1) a single dose cross-over study in T2DM, 2) a 14 day randomized, double-masked study in T2DM, 3) a 12-week randomized, double-masked, placebo-controlled study in obese subjects with normoglycemia (NG) or dysglycemia (DG; HbA1c: ≥5.7% and &amp;lt;10%). Results: LIK066 (2.5, 30, 300 mg single dose) significantly reduced glucose excursion and insulin secretion (p&amp;lt;0.01 at 30 and 300 mg) following oral glucose tolerance test (OGTT) in patients with T2DM (n=12), with the 300 mg dose completely suppressing glucose and insulin excursions. In patients with T2DM (n=30), LIK066 15 mg qd for 14 days reduced blood glucose (41 mg/dL by continuous glucose monitoring) and increased 24-hour urinary glucose excretion (100 g/d on day 14). Furthermore total GLP-1 and PYY increased (AUC0-3hrs &amp;gt;1.5-fold; p&amp;lt;0.05) post-OGTT, while GIP levels were reduced (&amp;gt;50%; p&amp;lt;0.05). In obese subjects (n=88), LIK066 150 mg qd for 12 weeks reduced weight by 5.70% (p&amp;lt;0.001) compared to placebo, with higher effects in DG (6.85%) vs. NG (4.55%). Additionally multiple metabolic parameters improved (reduced waist circumference, postprandial glucose and insulin, and increased postprandial glucagon). LIK066 was generally safe and well tolerated. The most common AEs were headache, flatulence and diarrhea. Conclusions: LIK066 treatment leads to favorable changes in a variety of metabolic hormones (increased GLP-1, PYY and glucagon, reduced GIP, glucose and insulin), all contributing to WL effects of LIK066. The dual inhibition of SGLT1/2 in both the gut and kidneys is an attractive strategy for obesity or diabetes treatment. Disclosure Y. He: None. W.G. Haynes: Employee; Self; AstraZeneca. C.D. Meyers: Employee; Self; Janssen Research &amp; Development. A. Amer: Employee; Self; Novartis Pharmaceuticals Corporation. Stock/Shareholder; Self; Novartis AG. Y. Zhang: Employee; Self; Novartis Pharmaceuticals Corporation. P.C. Mahling: None. A.E. Mendonza: None. S. Ma: None. W. Chutkow: Employee; Self; Novartis Pharmaceuticals Corporation. E.S. Bachman: None.

Relationship between glycemic control and OPG gene polymorphisms with lower bone mineral density in patients with type 1 Diabetes mellitus

ABSTRACT The aim of the present study was to investigate the bone mineral density (BMD) of patients with type 1 Diabetes mellitus (T1DM). We also assessed the association between osteoprotegerin (OPG) genetic polymorphisms and BMD. Genotyping was performed for 1181G>C and 163A>G OPG polymorphisms by allelic discrimination in 119 patients with T1DM and 161 normoglycemic (NG) individuals, aged 6 to 20 years old. Glycemic control, serum parameters of bone metabolism and BMD were evaluated. T1DM patients showed low BMD, poor glycemic control and decreased total calcium values when compared to controls (p < 0.05). For all the polymorphisms studied, the genotype and allele frequencies in patients with T1DM were not significantly different from the controls. In patients with T1DM, carriers of OPG 1181CC showed higher concentrations of ionized calcium compared to patients with GG+GC genotypes. These results suggest that low BMD is associated with poor glycemic control in T1DM. Despite the lack of a detected association between OPG polymorphisms and BMD in these patients, the increased ionized calcium in those carrying OPG 1181CC suggests a possible increase in osteoclastogenesis, a conclusion that may be supported by the lower BMD observed in these subjects.

Ethyl pyruvate attenuates myocardial ischemia-reperfusion injury exacerbated by hyperglycemia via retained inhibitory effect on HMGB1

BackgroundHyperglycemia (HG) exacerbates myocardial ischemia/reperfusion (I/R) injury and renders protective strategies ineffective by amplified inflammatory response via enhanced high-mobility group box-1 (HMGB1) release. This study investigated the role of ethyl pyruvate (EP) against myocardial I/R injury under a clinically relevant HG condition. MethodsSprague-Dawley rats (n=76) were randomly assigned to 6 groups: normoglycemia (NG)-Sham, NG-I/R-control (C, saline), NG-I/R-EP treatment (50mg/kg) upon reperfusion, HG-Sham, HG-I/R-C, and HG-I/R-EP treatment upon reperfusion. HG was induced by 1.2g/kg dextrose. I/R was induced by ligation of the left anterior descending artery for 30min followed by 4h of reperfusion. ResultsHG resulted in exacerbation of myocardial infarct size by 19% with amplified activation of HMGB1–receptors of advanced glycation end products/toll like receptors–NF-κB pathway compared to NG following I/R, which all could be attenuated by EP. EP treatment was associated with diminished tumor necrosis factor-α, interleukin-1β, and interleukin-6 expressions. It also served to normalize the increase in pro-apoptotic Bax and the decrease in anti-apoptotic Bcl-2 protein levels. These effects were associated with decreased myocardial apoptosis and infarct size (by 30% and 36% in the NG and HG groups, respectively) regardless of the glycemic condition. ConclusionHG exacerbated myocardial I/R injury through amplified inflammatory response via increased HMGB1 level. EP treatment upon reperfusion conveyed significant myocardial protection against the I/R injury under both NG and HG conditions. Common to both glycemic conditions, associated mechanisms involved attenuated increase in HMGB1 level and suppression of its down-stream pathways.

Ethyl pyruvate is renoprotective against ischemia-reperfusion injury under hyperglycemia

BackgroundHyperglycemia (HG) is common in cardiovascular surgeries due to diabetes, inflammation, and the neuroendocrine stress response. HG aggravates renal ischemia-reperfusion (I/R) injury through an increased inflammatory response, and blunts the protective effect of various measures. Ethyl pyruvate (EP) provides anti-inflammatory effects against I/R injury via inhibition of high-mobility group box 1 protein (HMGB1) release. This study aimed to determine the renoprotective effect of EP against I/R injury under HG. MethodsSprague–Dawley rats were randomly assigned at random to 8 groups: normoglycemia (NG)-sham, NG-I/R-control, NG-EP-I/R (pretreatment), NG-I/R-EP (posttreatment), HG-sham, HG-I/R-control, HG-EP-I/R, and HG-I/R-EP. Renal I/R was induced by 45 minutes of ischemia (clamping of renal arteries), followed by 24 hours of reperfusion. EP (50 mg/kg) was administered intraperitoneally at 1 h before ischemia (pretreatment) or on reperfusion (posttreatment). ResultsI/R injury under HG significantly aggravated the degree of renal tubular apoptosis and damage compared with the NG groups, which could be attenuated by both pretreatment and posttreatment of EP. I/R-induced increases in HMGB1 and Toll-like receptors (TLRs), activation of NF-kB, and resultant alterations in interleukin-1β, tumor necrosis factor-α, proapoptotic Bax, and antiapoptotic Bcl-2 were all favorably modulated by EP treatment in both the NG and HG groups compared with their corresponding control groups. ConclusionsDespite aggravation of renal I/R injury by HG through amplified inflammation, EP administration showed similar suppression of the HMGB1-TLR-NF-kB pathway in the HG and NG groups. EP retained anti-inflammatory, antiapoptotic, and renoprotective effects in the HG groups, whether administered before ischemia or on reperfusion.

221 - Regulation of β-TrCP Mediated Nrf2 Degradation by GSK3β during Hyperglycemic Renal Toxicity

Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a master regulator of cellular homeostasis that regulates more than 1% human genes including the ones related to glucose metabolism. Type 2 diabetes mellitus is associated with reduced Nrf2 mediated responses due to its compromised protein stability. The molecular mechanism behind Nrf2 degradation during hyperglycemia remains incompletely understood. Present study demonstrates that phosphorylation of Nrf2 by GSK3β at Neh6 domain causes its nuclear destabilization and ubiquitination by an E3 ligase β-TrCP, resulting in its loss of action in a keap1 independent manner. Co-immuno-precipitation studies and dual staining microscopy showed higher β-TrCP –Nrf2 interaction leading to its ubiquitination and degradation in the diabetic rats. Down-regulation of GSK3β by lithium chloride in vivo significantly decreased β-TrCP levels and enhanced Nrf2 nuclear stability compared to diabetic rats. In addition, nuclear Nrf2 retention upon GSK3β inhibition enhanced antioxidant levels and increased p-mdm2 (Ser 166) protein expression which suppressed p53 activation by its ubiquitination thereby preventing cell death. Renal tubular cells transfected with Nrf2 si-RNA and post treated with high glucose (HG) showed higher expression of Mst1, Bax and Bim pro-apoptotic proteins compared to normoglycemic (NG) cells suggesting that Nrf2 play a major role in precipitating diabetic kidney disease. Overall, the study confirms that hyperglycemia activates GSK3β (RNA/Protein) which in turn degrades Nrf2 via β-TrCP and limits cell survival. The data also shows potential role of GSK3β/ β-TrCP axis in Nrf2 instability during diabetes. Thus, targeted inhibition of GSK3β may be exploited to prevent the patho-complications evolved during hyperglycemic renal toxicity.