Basal Blood Glucose Levels Research Articles

8353 Impact of a plant-based protein diet on bone health and body composition in Sprague Dawley rats

Abstract Disclosure: M.M. Gomes: None. I.M. de Araújo: None. S.V. Rodriguez: None. F.R. Jorge: None. F.J. de Paula: None. Introduction:Vegetarian and vegan diets are associated with some health benefits; this has contributed to their growing popularity. Although studies remain limited, there is evidence that these patients may be at greater risk of reduced bone mineral density (BMD) and increased fracture risk. The general objective of this study was to analyze the impact of a high-protein, plant-based diet on bone tissue and body composition in male Sprague Dawley rats. Methodology: The study was approved by the Animal Ethics Committee. This study was done with 10 rats in the control group (CG), which received a standard AIN-93M diet; 11 in the normoprotein vegetarian group (NVG), with a normoprotein AIN-93 diet modified with pea protein and methionine; and 11 in the high-protein vegetarian group (HVG), with a modified AIN-93 diet with 40% pea protein and methionine. The dietary intervention lasted 8 weeks. The weight of the animals was measured weekly. Basal blood glucose levels were measured at baseline. At the end of the study, 6 rats from the CG, 6 rats from the NVG, and 5 rats from the HVG were subjected to a glucose tolerance test (GTT). The dual-energy X-ray absorptiometry (DXA) technique was applied in the baseline and final periods of the experiment to evaluate body composition, bone mineral content (BMC) and BMD. After sacrifice, bone strength was measured via a three-point flexion mechanical test on the tibia; brown adipose, epididymal, and retroperitoneal tissues, as well as the liver, were weighed on an analytical balance. Blood was used to measure circulatory levels of insulin, CTX, and P1NP (n=6/group). Results: Basal weight was similar between the groups, but the final weight was lower in the HVG as compared to the CG. In the DXA evaluation, the final lean mass was not different between the groups. On the other hand, total fat mass was lower in the HVG than in the CG and the NVG and the fat percentage was lower in the HVG than in the CG. The weight of epididymal adipose tissue was lower in the HVG than in the NVG, but retroperitoneal and brown adipose tissues were lower in HVG than CG. Liver weight was similar between groups. BMD was similar between groups; however, BMC was lower in the HVG than in the CG and the NVG. Bone strength was similar between groups. There was no difference in glycemia at baseline or during GTT at the eighth week. Serum levels of insulin, CTX, and P1NP were similar between groups. Discussion and Conclusion: A normoprotein vegetarian diet that includes all essential amino acids appears to have no negative impact on body composition or BMD in rats. On the other hand, a high-protein diet with plant protein appears to have a negative impact on bone mineral content, in addition to reducing the amount of body fat and weight. This finding expands on previous results by showing that high-protein diets have a negative impact on bone even when the protein is of plant origin. Presentation: 6/1/2024

Effect of Hafnia alvei on Morphophysiologic Parameters and Gut Microbiota of Mice with Inherited Type 2 Diabetes Mellitus.

Inbred mouse strains KK.Cg-a/a and KK.Cg-Ay/a known as genetic models of type 2 diabetes mellitus significantly surpassed the control strain C57BL/6J in the body weight, relative weight of extractable fat, and basal blood glucose levels. Real-timePCR of fecal samples from KK.Cg-a/a and KK.Cg-Ay/a mice revealed dysbiosis typical of type 2 diabetes mellitus in humans and animals. Long-term intragastric administration of a suspension of Hafnia alvei bacteria had no effect on the above morphometric and biochemical parameters. At the same time, recovery of the Bacteroides spp. population in KK.Cg-Ay/a mice and a decrease in the number of Bifidobacterium spp. in KK.Cg-a/a mice were observed. The possibility of therapeutic use of the probiotic based on H. alvei is discussed.

Pre-Existing Metabolic Conditions Impair the Beneficial Effects of Estrogen on Memory and Cognition in Ovariectomized Mice

Loss of estrogen, as a result of menopause, has been associated with cognitive decline and memory loss. Intriguingly, studies conducted with postmenopausal healthy women suggest that hormone replacement therapy improves cognitive functions. In contrast, clinical data involving subjects with a wide range of health conditions indicate that the effect of estrogen treatment varies. In addition to the cognitive decline, postmenopausal women have an increased risk of developing metabolic disorders including obesity, insulin resistance, and diabetes. In this study, we tested the hypothesis that pre-existing metabolic conditions reduce the beneficial effects of estradiol treatment and are responsible for the conflicting findings in cognitive functions after menopause. Female, seven-month-old C57BL/6J mice were fed with either a high-fat diet (HFD, BioServ #F3282) or a control diet (LFD, BioServ #F4031) for 10 to 14 weeks, then ovariectomized (OVX). In a randomized manner, mice received a silastic tube implant containing either 17β-estradiol (E2) or vehicle (VEH, cholesterol) and stayed on their respective diets. Body composition and metabolic parameters were measured in the four groups (HFD+VEH, HFD+E2, LFD+VEH, LFD+E2) to assess metabolic status, and novel object recognition test was performed to assess memory and cognition. Before OVX, HFD fed mice had significantly increased body weight, fat mass, basal blood glucose and insulin levels compared to mice fed with LFD. After OVX, LFD-VEH mice developed obesity and had elevated blood glucose and insulin levels, whereas E2 treatment prevented the metabolic changes. Interestingly, HFD-VEH mice had a reduction in fat mass after OVX, which was further decreased with E2 treatment. Overall, mice treated with E2 had lower body weight, fat mass, blood glucose and insulin levels compared to VEH treated mice regardless of their diet, and HFD fed mice had a higher body weight than mice fed the LFD, irrespective of their treatment. The novel object recognition test revealed that LFD-E2 mice spent significantly more time exploring the novel object than LFD-VEH mice. In contrast, there was no difference in exploration time between HFD-VEH and HFD-E2 treated mice. These preliminary results suggest that 1) E2 treatment may be beneficial to prevent the development of metabolic changes and improve cognitive function in LFD fed mice; 2) OVX alone did not worsen the metabolic status of diet-induced obese mice; 3) E2 treatment partially improved the metabolic status of HFD fed mice by decreasing fat mass, blood glucose and insulin levels; and 4) E2 treatment did not improve memory and cognition of HFD fed mice. Source of funding: P01AG071746 8456. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Stress hyperglycemia increases short-term mortality in acute ischemic stroke patients after mechanical thrombectomy

Background and purposeGlucose-to-glycated hemoglobin ratio (GAR) is considered a more reliable marker of stress hyperglycemia by correcting for basal blood glucose levels. This study aimed to investigate the extent to which GAR is associated with 3 month and 1 year all-cause mortalities in patients with acute ischemic stroke (AIS) undergoing mechanical thrombectomy (MT).MethodsWe retrospectively followed 553 AIS patients who underwent MT. The degree of stress hyperglycemia was quantified as the GAR, defined as fasting plasma glucose (mmol/L)/hemoglobin A1c (HbA1c) (%) on the second day after admission. According to the GAR quartiles, the patients were further categorized into four groups (group 1-group 4). We assessed the association between GAR and all-cause mortalities, clinical outcomes during hospitalization and function outcomes at 3 months. The associations between stress hyperglycemia and all-cause mortalities were analyzed using a Cox proportional-hazards model, while other outcomes were analyzed using multiple logistic regression analysis.ResultsThe follow-up lasted a median of 18 months (range 0–66 months). The 3 month mortality rate was 9.58% (n = 53) and the 1 year mortality rate was 18.62% (n = 103). The Kaplan–Meier analysis revealed a significant inverse relationship between GAR and mortality (P < 0.001). In the Cox proportional-hazards model at 3 months, compared with group1, group 4 of GAR was associated with a significant increase in the risk of 3 month mortality (hazard ratio [HR] = 4.11, 95% confidence interval [CI] 1.41–12.0, P = 0.01) after adjusting for potential covariates. On multivariate logistic regression analysis, GAR was strongly associated with an increased risk of 3 month poor function outcome.ConclusionsStress hyperglycemia, quantified by a higher GAR, is associated with all-cause mortality and poor functional outcomes in patients with AIS who undergo MT. Furthermore, GAR may contribute to improving the predictive efficiency of all-cause mortality in patients with AIS after MT, especially short-term all-cause mortality.

Effects of metformin on behavioral alterations produced by chronic sucrose consumption in male rats.

Excessive consumption of sugary drinks negatively impacts the developing brain, producing long-lasting behavioral and metabolic disorders. Here, we study whether treatment with the antihyperglycemic agent metformin prevents some of the anxiety and memory alterations produced by chronic sucrose consumption. Male Sprague-Dawley rats had unrestricted access to water (control group) and a bottle containing a 10% sucrose solution (sucrose group, SUC) for 35 days. In parallel, a group of animals from SUC received metformin (25 mg/kg or 50 mg/kg, orally; MET 25 and MET 50 groups, respectively). After 2 weeks of metformin treatment, the animals weighed less than controls. SUC and MET 50 groups compensated for the caloric intake from the sugary solution by consuming less chow. In contrast, total energy intake in MET 25 was higher than the rest of the groups, but they still weighed less than control and SUC groups, suggesting that at this concentration, metformin delays body growth. The animals were then tested for the open field (OF), elevated plus maze (EPM) and novel object location (NOL) tests. In the OF, SUC animals spent more time in the central zone of the arena, evidenced by an increased number of entries and the distance traveled there. In the EPM, SUC animals spent more time in the open arms and less time in the central square. Metformin treatment prevented the decreased anxiety observed in SUC animals in the OF and EPM. In the NOL test, SUC animals showed less interest in novelty and metformin treatment did not improve this alteration. The preference for open spaces in the OF and EPM were associated with increased serum triglycerides (TG) and malondialdehyde levels in the medial prefrontal cortex (mPFC) and the hippocampus (HIP), while poor memory performance was associated with high basal blood glucose levels. In conclusion, the decreased anxiety-like behavior produced by chronic sucrose consumption was prevented by metformin treatment, through a mechanism that probably involves normalization of TG levels and decreased oxidative stress in mPFC and HIP.

Study on the Hyperglycemic Effect of GLP-1 in Spinibarbus denticulatus by Oral Administration and Intraperitoneal Injection Methods.

Glucagon-like peptide-1 (GLP-1), one of the expression products of the proglucagon (pg) gene, is an incretin mainly secreted by the gastrointestinal system. In mammals, GLP-1 has hypoglycemic and food-inhibiting effects; while in some fish species, it has been confirmed to increase blood glucose by promoting gluconeogenesis and stimulating glycogenolysis. In order to more deeply understand the role of GLP-1 in the process of glycometabolism in herbivorous fish, the pg gene was cloned from Spinibarbus denticulatus to obtain its sequence characteristics, and the changes in blood glucose level and pg gene expression in S. denticulatus were further explored by feeding with three kinds of carbohydrates and intraperitoneal injection of GLP-1. Basal and temporal blood glucose levels and pg gene expression of S. denticulatus (91.68 ± 10.79 g) were measured at 0, 1, 3, 5, 7, and 12 h after oral administration (n = 4). Then, the changes of blood glucose levels and pg and glucokinase (gk) gene expressions of S. denticulatus (94.29 ± 10.82 g) were determined at 0, 30, 60, and 120 min after intraperitoneal injection (n = 4). It was shown that polysaccharides could induce the upregulation of pg gene expression faster than monosaccharides and stimulate the secretion of GLP-1 in the intestine. Intraperitoneal injection of GLP-1 peptide rapidly raised blood glucose levels, and pg gene expression in the anterior intestine, whole brain, and hepatopancreas decreased continuously after 30 minutes. These results showed that S. denticulatus might inhibit the excessive accumulation of blood glucose by reducing the expression of the pg gene and increasing the expression of gk gene in a short time. It was speculated that GLP-1 of S. denticulatus might have a "gut-brain-liver" pathway similar to mammals in glycemia regulation. Therefore, this study provided a novel perspective for explaining the functional differences of GLP-1 in herbivorous fish and mammals.

Blood glucose kinetics during concentric and eccentric muscle contractions

Studies comparing the two classes of stimuli (concentric and eccentric) have shown differences in the improvement of cardiovascular, metabolic, and muscle strength gain. This is an experimental, quantitative, and prospective study that aimed to verify the effect of eccentric exercise on glycolytic consumption and kinetics. The blood glucose kinetics of 17 male subjects was evaluated during a treadmill exercise with a 10% declined floor and velocity that required a 60% metabolic activity of VO2max, for 30 minutes. Seventy-two hours later, the same subjects exercised on the treadmill with a 10% inclined floor and 60% VO2max, for 30 minutes. To quantify glucose, blood samples were collected before the exercise, every three minutes along the 30 minutes of physical activity, and five and 10 minutes after finishing the exercise. For the downward slope, there was a homogeneous group behavior for blood glucose dynamics during the exercise, which was characterized by a monotonic decrease of glucose levels until reaching a minimum value at experimental times between 20 and 30 min, followed by a progressive recovery toward initial values. For the acclivity condition, blood glucose dynamics did not follow such a homogeneous behavior. A set of different types of dynamics could be identified. Experimental data showed that the type of dynamics could be predicted, to some extent, by the basal blood glucose level of subjects. The type of floor slope (upward or downward) directly affected glycolytic consumption and kinetics for the individuals analyzed.

Kynurenine-3-monooxygenase expression is activated in the pancreatic endocrine cells by diabetes and its blockade improves glucose-stimulated insulin secretion

Type 2 diabetes is associated with an inflammatory phenotype in the pancreatic islets. We previously demonstrated that proinflammatory cytokines potently activate the tryptophan/kynurenine pathway (TKP) in INS-1 cells and in normal rat islets. Here we examined: (1) the TKP enzymes expression in the diabetic GK islets; (2) the TKP enzymes expression profiles in the GK islets before and after the onset of diabetes; (3) The glucose-stimulated insulin secretion (GSIS) in vitro in GK islets after KMO knockdown using specific morpholino-oligonucleotides against KMO or KMO blockade using the specific inhibitor Ro618048; (4) The glucose tolerance and GSIS after acute in vivo exposure to Ro618048 in GK rats. We report a remarkable induction of the kmo gene in GK islets and in human islets exposed to proinflammatory conditions. It occurred prominently in beta cells. The increased expression and activity of KMO reflected an acquired adaptation. Both KMO knockdown and specific inhibitor Ro618048 enhanced GSIS in vitro in GK islets. Moreover, acute administration of Ro618048 in vivo improved glucose tolerance, GSIS and basal blood glucose levels in GK rats. These results demonstrate that targeting islet TKP is able to correct defective GSIS. KMO inhibition could represent a potential therapeutic strategy for type 2 diabetes.

Low molecular weight NGF mimetic GK-2 normalizes the parameters of glucose and lipid metabolism and exhibits a hepatoprotective effect on a prediabetes model in obese Wistar rats.

Signs of metabolic syndrome and prediabetes preceding type 2 diabetes are modelled in an experiment using a high-fat diet (HFD). The aim of this work was to study the effect of a low molecular weight systemically active nerve growth factor mimetic, compound GK-2 (hexamethylenediamide bis[N-monosuccinyl-L-glutamyl-L-lysine]), on indicators of abdominal obesity, basal blood glucose level, glucose tolerance, cholesterol and triglyceride blood levels, as well as the morphological structure of the liver in male Wistar rats fed a HFD. Rats were divided into three groups: one of them received standard food (control) and two others were fed a HFD containing 45% fat, 35% carbohydrates and 20% protein, with a total caloric value of 516 kcal/100 g, over 12 weeks. Starting from the ninth week, for the next 4 weeks, one of the HFD groups was treated orally with saline whilst the other group was treated orally with GK-2 at a dose of 5 mg/kg. GK-2 was found to reduce the basal glycaemia level and improve glucose tolerance, as well as to reduce the blood level of cholesterol by 30% and that of triglycerides by 28% in comparison with the saline-treated HFD animals. GK-2 reduced the degree of abdominal obesity to the level of the healthy animals and eliminated morphological abnormalities in the liver caused by the HFD. The results of the study determine the feasibility of further GK-2 research as a potential agent for prediabetes treatment.

196-OR: (Pro) renin Receptor Signaling in Tyrosine Hydroxylase Neurons in the Hypothalamic Paraventricular Nucleus Is Required for Obesity-Associated Glucose Metabolic Impairment

We recently reported that neuron-specific (pro) renin receptor (PRR) deletion reduces fasting blood glucose (BG) levels and improves glucose tolerance in high fat diet (HFD) -treated mice. To investigate underlying mechanisms, we focused on tyrosine hydroxylase (TH) neurons in the hypothalamic paraventricular nucleus (THPVN neurons) , a largely GABAergic population that expresses PRR and projects to autonomic centers important for BG regulation. To delete PRR from THPVN neurons, we injected into the PVN of PRR-loxp mice either an adeno-associated virus expressing cre recombinase under the control of the rat TH promoter (AAV2-TH-Cre) or AAV2-TH-eGFP as a control. After placing animals on a HFD for 6 wk, we found that the obesity-induced increase of fasting BG levels was blocked by PRR deletion from THPVN neurons (118.0 ± 3.7 vs. 139.3 ± 6.6 mg/dL, n=6/group, p= 0.04) , accompanied by a trend towards improved i.p. glucose tolerance (AUC: 12990 ± 1136 vs. 9467 ± 1298, n=6, p=0.07) . To further investigate the role of THPVN neurons in glucose homeostasis, we employed a Designer Receptors Exclusively Activated by Designer Drugs (DREADD) approach whereby AAV2-TH-Cre was injected bilaterally into the PVN of either hM3Dq-loxp (excitatory) or hM4Di-loxp (inhibitory) mice, and continuous glucose monitoring was performed in conscious, freely moving mice. Following i.p. injection of clozapine-N-oxide (CNO, 1 mg/Kg) or vehicle, we found that whereas basal BG levels were not altered by THPVN neuron activation, THPVN neuron inhibition significantly increased basal BG levels compared to vehicle in male (ΔBG: 31.6 ± 8.6 mg/dL vs. 11.0 ± 6.7 mg/dL, n=4, p=0.03) but not female mice. These findings establish a role for THPVN neuron PRR signaling in the pathogenesis of obesity-associated glucose metabolic impairment, and show that in male mice, basal THPVN neuron activity is required for normal glucose homeostasis. Disclosure L.Ac. souza: None. C.J.Worker: None. A.Gayban: None. M.W.Schwartz: None. Y.Feng earley: None. Funding National Institutes of Health (R01HL122770, R35HL155008, RDK83042, RDK124238)

Effects of Adiponectin on Glucose Metabolism in the Hepatopancreas of Grass Carp (Ctenopharyngodon idella)

Adiponectin plays critical roles in the regulation of energy metabolism in mammals. However, its roles in fish remain to be evaluated. In this study, we examined the tissue distribution characteristics of two adiponectin genes, namely, adipoqa and adipoqb, at transcriptional level, and detected the changes of their mRNA expressions during oral glucose tolerance test (OGTT). Furthermore, we prepared recombinant proteins of grass carp adiponectin A and adiponectin B and studied their roles in the regulation of glucose metabolism in the hepatopancreas of grass carp in vivo. The results showed that adipoqa mRNA was mainly expressed in the heart of grass carp, followed by red muscle, white muscle, and adipose tissue, while adipoqb mRNA was highly expressed in adipose tissue, red muscle, and white muscle. The mRNA levels of adipoqa and adipoqb in the white muscle and heart were significantly decreased at 1 h post OGTT, and their expressions in the heart were also suppressed at 3 h, whereas the transcriptional levels of adipoqa and adipoqb in the red muscle were elevated significantly at 12 h after glucose load. Intraperitoneal injection of adiponectin A or adiponectin B reduced basal serum glucose levels of grass carp and caused an increase of glycogen content in the hepatopancreas. Adiponectin A upregulated the expression of enzyme genes related to glycolysis (gk and pk) and glycogen synthesis (gys) in the hepatopancreas, and adiponectin B promoted the expression of pk and gys, as well as g6pase in the glycogenetic pathway. Besides, adiponectin A and adiponectin B have opposite regulatory effects on glut2 mRNA expression. These results suggested that adiponectin A and adiponectin B promoted glycolysis, gluconeogenesis, and glycogen synthesis, increased glycogen content of hepatopancreas, and reduced basal blood glucose level, and they may have an antagonistic regulatory mechanism in regulating glucose uptake.

Absence of the mitochondrial translocator protein 18kDa in mice does not affect body weight or food intake responses to altered energy availability.

Changes in mitochondrial function in a variety of cells/tissues are critical for orchestrating systemic energy homeostasis and are linked to the development of obesity and many of its comorbidities. The mitochondrial translocator protein of 18kDa (TSPO) is expressed in organs throughout the body, including the brain, liver, adipose tissue, gonads and adrenal glands, where it is implicated in regulating steroidogenesis and cellular metabolism. Prior work from our group and others has shown that, in rodents, TSPO levels are altered in adipose tissue by obesity and that modulation of TSPO activity may impact systemic glucose homeostasis. Furthermore, in vitro studies in a variety of cell types have implicated TSPO in mediating cellular energetics and substrate utilisation. Although mice with germline global TSPO deficiency (TSPO-/- ) have no reported changes in body weight under standard husbandry conditions, we hypothesised that, given the roles of TSPO in regulating mitochondrial function and cellular metabolic flexibility, these animals may have alterations in their systemic response to altered energy availability, either nutritional excess or insufficiency. In agreement with published work, compared to wild-type (TSPO+/+ ) littermates, TSPO-/- mice of both sexes did not exhibit differences in body weight on standard chow. Furthermore, following a 12-hour overnight fast, there was no difference in weight loss or compensatory food intake during re-feeding. Five weeks of feeding a high-fat diet (HFD) did not reveal any impact of the absence of TSPO on body weight gain in either male or female mice. Basal blood glucose levels and glucose clearance in a glucose tolerance test were influenced by feeding a HFD diet but not by genotype. In conclusion, in the paradigms examined, germline global deletion of TSPO did not change the physiological response to deviations in systemic energy availability at the whole organism level.

Safety assessment of the dried hydroethanolic extract of Pterocarpus erinaceus Poir. stem bark

Background: Medicinal plants are used to treat diseases but can be toxic in certain doses or due to the presence of certain compounds. Thus, the safety of medicinal plants must be scientifically established before their use. Pterocarpus erinaceus is one of medicinal plants used in traditional medicine to treat several diseases including diabetes and its complications.Purpose: This study aims to assess the safety of P. erinaceus in vitro in brine shrimp larvae Artemia salina and in vivo in Sprague Dawley rats in order to prove its use in the formulation of phytomedicine.Study design and methods: The fresh bark of P. erinaceus stem was dried and powdered. The dry extract was obtained by hydroethanolic extraction. The cytotoxicity of P. erinaceus was evaluated in vitro in brine shrimp larvae Artemia salina for 24 h. The acute and subchronic toxicity of P. erinaceus were assessed in vivo in rats by oral administration of extract at a single dose of 5 g/kg bw and repeated doses of 500 and 1000 mg/kg bw for 28 consecutive days.Results: The extract of the stem bark of P. erinaceus has a LC50 = 2.941 mg/mL in brine shrimp larvae Artemia salina and considered nontoxic on the Mousseux scale. In acute toxicity using ATC method, a single oral administration of extract did not result in any mortalities or signs of toxicity in gross necropsy during the 14 days of observation in rats. Therefore, P. erinaceus has a LD50 > 5 g/kg bw and considered nontoxic. Subchronic oral administration of P. erinaceus at doses of 500 and 1000 mg/kg bw, also did not cause any mortalities or significant changes in body weight, relative weight of organs, hematological and biochemical parameters. No abnormalities in liver, spleen and kidneys were observed in the histopathological study. However, a significant (p < 0.001) decrease in basal blood glucose levels in rats was observed during treatment, but was not correlated with organ damage.Conclusion: This study showed that P. erinaceus stem bark was safe and had no cyto-histopathological impact in animal health. Thus, it can be used in the formulation of phytomedicine; but further studies such as genotoxicity and chromosomal aberration are needed to prove its safety at the genomic level.

Glucose Variability and Risk of Hepatocellular Carcinoma in Patients with Diabetes: A Nationwide Population-Based Study.

Although diabetes is a well-known risk factor for hepatocellular carcinoma, exactly which metabolic parameters of diabetes are associated with hepatocellular carcinoma remain unexplored. In this study, we investigated the relationship between glucose variability (GV) and hepatocellular carcinoma in patients with diabetes through a nationwide population-based study. A population-based cohort study including 674,178 diabetic subjects participating in more than three health examinations within 5 years from the index year (2009 and 2010) were followed until the end of 2017. The coefficient of variation, SD, variability independent of the mean, and average real variability were calculated as GV indices. During a median follow-up of 6.7 years, there were 5,494 cases of hepatocellular carcinoma. When groups were classified according to glucose level, the highest risk for hepatocellular carcinoma was observed when the basal blood glucose level was 180 mg/dL or greater [adjusted HR (aHR), 1.19; 95% confidence interval (CI), 1.08-1.31]. We observed increasing trends for the relationship between GV and hepatocellular carcinoma in multivariable Cox proportional analyses. The risk of hepatocellular carcinoma increased by 27% (aHR, 1.27; 95% CI, 1.17-1.38) for the highest quartile of GV relative to the lowest quartile. These findings were consistent regardless of the presence of chronic viral hepatitis or cirrhosis, alcohol consumption, or body mass index. GV was an independent predictor of hepatocellular carcinoma, even after adjusting for confounding factors. There was a linear relationship between increase in GV and prevalence of hepatocellular carcinoma. Visit-to-visit GV might be helpful for identifying patients with diabetes at high risk of hepatocellular carcinoma.

Admission blood glucose level as an ischemic stroke risk modifier in patients with new-onset non-valvular atrial fibrillation

Abstract Background Several scores have been proposed to assess the stroke risk in patients with non-valvular atrial fibrillation (NVAF). However, type 2 diabetes mellitus (T2DM) is considered a major stroke risk factor regardless of glycemic control. Whether basal blood glucose level modifies the risk of stroke in NVAF is still unclear. Purpose To evaluate the risk of ischemic stroke according to the presence of T2DM and admission blood glucose (ABG) level in patients with new-onset NVAF starting direct oral anticoagulants (DOACs). Methods We analyzed all consecutive patients with NVAF at our outpatient clinic from January to December 2018. The study population was constituted by 1014 patients with new-onset NVAF starting DOACs. Baseline characteristics were evaluated in the overall cohort whereas outcomes were assessed for 915 patients. The median follow-up time was 19.6±12.9 months. Results Overall, 50.3% were male with a mean age of 73.9±12.5 years. Diabetic NVAF patients were more frequently male (p=0.04) with higher prevalence of dyslipidemia (p&amp;lt;0.001), hypertension (p&amp;lt;0.001), severe renal impairment (p=0.02), peripheral vasculopathy (p=0.007) and history of myocardial infarction (p&amp;lt;0.001) compared to non-diabetic NVAF. Conversely, no differences were observed between subgroups in terms of age (p=0.8). Baseline blood glucose level was significantly higher in the diabetic NVAF population (160±67 mg/dL vs 119±39 mg/dL; p&amp;lt;0.001). As expected, the mean CHA2DS2-VASc score was significantly higher in diabetic NVAF compared to non-diabetic group (4.7±1.4 vs 3.2±1.5; p&amp;lt;0.001). During a 2 year-follow up period, we collected 27 (3.0%) ischemic stroke. As expected, the rates of stroke were significantly higher in diabetic NVAF (7.6% vs 2.3%, p&amp;lt;0.001). Also, the ABG was significantly greater in NVAF who had an ischemic stroke compared to others (160±68 mg/dL vs 119±39 mg/dL, p=0.005). The incidence of stroke was almost five-time greater in NVAF with ABG level major than 150 mg/dl (9.8% vs 1.9%, p&amp;lt;0.001). At multivariate Cox-regression model adjusted for age, sex and presence of T2DM, blood glucose level at admission was the only independent predictor of ischemic stroke at follow up (HR 1.01, 95% CI 1.001–1.02; p=0.03). Finally, another multivariate Cox-regression model, adjusted for the mean CHA2DS2-VASc score, showed that the ABG level still remained a strong independent predictor of ischemic stroke at follow up (HR 1.012, 95% CI 1.003–1.02; p=0.01). Conclusions Diabetic NVAF had a worse baseline profile and higher stroke risk compared to non-diabetic NVAF. Baseline blood glucose level was an independent predictor of stroke regardless of the presence of T2DM or stroke risk profile. These findings underline the role of basal blood glucose level as a potential stroke risk modifier and therefore emphasize the importance of its routine determination to better stratify the stroke risk in NVAF starting DOACs. Funding Acknowledgement Type of funding source: None

Humanized GPRC6AKGKY is a gain-of-function polymorphism in mice

GPRC6A is proposed to regulate energy metabolism in mice, but in humans a KGKY polymorphism in the third intracellular loop (ICL3) is proposed to result in intracellular retention and loss-of-function. To test physiological importance of this human polymorphism in vivo, we performed targeted genomic humanization of mice by using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9) system to replace the RKLP sequence in the ICL3 of the GPRC6A mouse gene with the uniquely human KGKY sequence to create Gprc6a-KGKY-knockin mice. Knock-in of a human KGKY sequence resulted in a reduction in basal blood glucose levels and increased circulating serum insulin and FGF-21 concentrations. Gprc6a-KGKY-knockin mice demonstrated improved glucose tolerance, despite impaired insulin sensitivity and enhanced pyruvate-mediated gluconeogenesis. Liver transcriptome analysis of Gprc6a-KGKY-knockin mice identified alterations in glucose, glycogen and fat metabolism pathways. Thus, the uniquely human GPRC6A-KGKY variant appears to be a gain-of-function polymorphism that positively regulates energy metabolism in mice.

Western Diet Promotes Renal Injury, Inflammation, and Fibrosis in a Murine Model of Alström Syndrome

Introduction: Alström syndrome is a rare recessive genetic disease caused by mutations in ALMS1, which encodes a protein that is related to cilia function and intracellular endosome trafficking. The syndrome has been linked to impaired glucose metabolism and CKD. Polymorphisms in Alms1 have likewise been linked to CKD, but little is known about the modification of the phenotype by environmental factors. Methods: To gain further insights, the fat aussie (foz) mouse strain, a genetic murine model of Alström syndrome, was exposed to a normal chow (NC) or to a Western diet (WD, 20% fat, 34% sucrose by weight, and 0.2% cholesterol) and renal outcomes were measured. Results: Body weight and albuminuria were higher in foz than in wild-type (WT) mice on both diets but WD significantly increased the difference. Measurement of plasma creatinine and cystatin C indicated that glomerular filtration rate was preserved in foz versus WT independent of diet. Renal markers of injury, inflammation, and fibrosis were similar in both genotypes on NC but significantly greater in foz than in WT mice on WD. A glucose tolerance test performed in foz and WT mice on WD revealed similar basal blood glucose levels and subsequent blood glucose profiles. Conclusions: WD sensitizes a murine model of Alström syndrome to kidney injury, inflammation, and fibrosis, an effect that may not be solely due to effects on glucose metabolism. Polymorphisms in Alms1 may induce CKD in part by modulating the deleterious effects of high dietary fat and sucrose on kidney outcome.

1758-P: Type 2 Diabetes–Associated Mood Disorders: Role of Insulin Resistance in Serotonergic Neurons

Type-2 Diabetes mellitus (T2DM) is mainly characterized by insulin-resistance and often accompanied by comorbidities such as major depressive disorders (MDD). Indeed, patients with T2DM are twice more likely to suffer from MDD and clinical studies have shown that insulin resistance is positively correlated with the severity of depressive symptoms. In preclinical studies, we and others showed that mice fed a High Fat Diet (HFD) for 16 weeks display both hallmarks of T2DM including increased basal blood glucose level and impaired glucose tolerance and anxio-depressive-like behaviors. We also showed that the insulin-sensitizing agent metformin reverses HFD-induced impaired glucose homeostasis and emotional alterations. As the serotonergic (5-HT) system in the brain is one of the major neuronal networks involved in the pathophysiology of MDD, we hypothesized that insulin resistance at the level of 5-HT neurons contributes to the development of mood disorders in a context of T2DM. Using patch-clamp electrophysiology, we show that HFD feeding impairs the excitability of dorsal Raphe 5-HT neurons while this effect is reversed by metformin treatment. Our data also show that local application of insulin increases firing rate of 5-HT neurons in chow fed animals and this is consistent with our histochemical observations indicating that the dorsal Raphe is endowed with a high density of insulin positive cells. Additionally, preliminary data suggest that intranasal or intra-Raphe insulin injection decreases anxiety in the elevated plus maze test. Our current work aims at evaluating the intranasal and intra-Raphe insulin injection in mice lacking the insulin receptor specifically in 5-HT neurons on emotional behaviors. Altogether, this study suggests that insulin action onto dorsal Raphe 5-HT neurons is important to control emotional behaviors and that insulin-resistance in these neurons is critical in the development of T2DM-associated mood disorders. Disclosure H. Martin: None. S. Bullich: None. F. Ducrocq: None. M. Graland: None. C. Olivry: None. S. Laye: None. B. Guiard: None. X. Fioramonti: None. Funding Société Francophone du Diabète; INRAE-Région Nouvelle Aquitaine

Antihyperglycemic activity of total extract and fractions of Anogeissus leiocarpus

Anogeissus leiocarpus (Combretaceae) is a medicinal plant used by traditional practitioners to treat people living with diabetes mellitus in Togo. The objective of this work was to evaluate the hypoglycemic activity of the plant. The hydroalcoholic extract and fractions of A. leiocarpus roots was evaluated on hyperglycemic mice by oral glucose tolerance test (OGTT) and on normoglycemic mice. The effect of the total extract and fractions was also measured on the adsorption and absorption of glucose respectively in vitro and ex vivo. At 30 minutes after glucose overload, the total extract and all fractions (supernatant and pellet) significantly (p&lt; 0.0001) reduced hyperglycemia compared to controls. However, this reduction in hyperglycemia was greater in mice treated with the supernatant fraction at 100 mg.kg-1 during 180 minutes (p &lt; 0.01). In our conditions, the extract at the 500 mg.kg-1 of the total extract in normoglycemic mice did not significantly decrease (p&gt;0.05) basal blood glucose levels compared to controls. In vitro, the extract and fractions of A. leiocarpus adsorbed glucose and inhibited glucose intestinal absorption ex vivo. The results of this study support the use of A. leiocarpus as an antidiabetic plant.&#x0D; Keywords: Anogeissus leiocarpus- fractions- antihyperglycemic- adsorption – absorption.

Resident bacteria in the small intestine can shape host drug metabolism through innate lymphoid cell‐derived IL‐22

The ability of the intestinal microbiota to influence drug metabolism has been recognized, however the mechanisms through which this occurs remain relatively unexplored. Recent work in germ‐free mice has demonstrated that conventionalization with specific pathogen free (SPF) microbiota influences the expression of cytochrome P450 (CYP) enzymes in the liver and small intestine (SI), two important sites for drug metabolism. Given that CYP enzymes, including CYP3A11 in mice, account for roughly 75% of total drug metabolism, we sought to investigate the role of specific gut‐residing microbes in shaping the expression and activity of these enzymes. Utilizing PCR arrays, we found that colonization of SPF mice with the immunomodulatory bacteria segmented filamentous bacteria (SFB) altered the expression of various CYP enzymes in the SI (but not liver), with Cyp3a11 being one of the most downregulated genes. Further analysis showed that SFB‐induced IL‐22 production by type 3 innate lymphoid cells (ILC3) correlated with reduced SI Cyp3a11 expression. Additionally, SFB colonization had no effect on the expression of Cyp3a11 in the SI of IL‐22−/− mice or RAG1−/− mice in which ILCs were depleted with anti‐Thy1 antibody. Both SFB colonization and administration of IL‐23, to activate IL‐22 from ILC3, increased the ability of the CYP3A11‐metabolized drug glibenclamide to decrease basal blood glucose levels when administered orally. In mouse SI enteroid cultures, IL‐22 dose‐dependently reduced the expression of Cyp3a11 and decreased the ability of SI enteroids to metabolize CYP3A11 specific substrates, effects that were blocked by the STAT3 inhibitor Stattic. Finally, when applied to human ileal organoids, IL‐22 induced robust changes in the transcriptome, with substantial effects on drug metabolism pathways centred around CYP3A. Our data suggest that specific gut‐resident microbes can influence the expression and activity of the drug metabolizing enzyme CYP3A11. This involves the production of IL‐22 by small intestinal ILC3, which down regulates CYP3A11 in the SI epithelium. These findings provide an understanding of how the intestinal microbiota modulates host drug metabolism, and how microbiota‐immune cell crosstalk may influence the efficacy and toxicity of various drugs.Support or Funding InformationAlberta Innovates, Canadian Association of Gastroenterology, Canadian Institute of Health Research