Impaired Oral Glucose Tolerance Test Research Articles

OR35-02 Novel Androgen Receptor PROTAC Degrader ARV-110 Ameliorates Androgen Mediated Renal Injury in a Mouse Model of Polycystic Ovary Syndrome

Abstract Disclosure: J. Basnet: None. S. Rezq: None. A.M. Huffman: None. T.E. Asala: None. L.L. Yanes Cardozo: None. D.G. Romero: None. Background: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in reproductive-age women. PCOS women often have increased central adiposity, insulin resistance, and renal injury. Hyperandrogenism is prevalent in PCOS women and is associated with renal injury and dysfunction. Androgens bind to the androgen receptor (AR) to impart downstream transcriptional activity. Targeting the AR may alleviate PCOS complications; however, the use of AR blockers is limited by their severe adverse effects in females; thus, safer alternative approaches are highly needed. The novel AR Proteolysis Targeting Chimera (PROTAC) degrader ARV-110 has shown high efficacy to prevent AR signaling in prostate cancer; however, its efficacy in PCOS is unknown. Using a well-established hyperandrogenemic mouse model of PCOS, we aim to test the hypothesis that decreasing AR protein levels with ARV-110 ameliorates androgen-mediated derangements in PCOS. Methods: Three-week old female mice (C57BL/6N) were implanted with Silastic tubes filled with the androgen dihydrotestosterone (DHT, 8mg, s.c., 90 days) or vehicle (n=6/grp). Four weeks post-DHT administration, mice were treated with ARV-110 (1 mg/kg/day, s.c.) for an additional 8 weeks. Body composition (EchoMRI) and oral glucose tolerance test (OGTT) were assessed. Kidney weight was measured by gravimetry. AR, extracellular matrix (ECM) remodeling marker MMP9, pro-fibrotic marker TGF-β, renal injury markers (urinary albumin to creatinine ration (UACR), KIM1, and NGAL) were assessed by Western-blot or ELISA. Results: DHT mice showed significant (p<0.05) increases in body weight (27.3±0.6 vs 23.9±0.26 g), fat mass (4.2±0.6 vs 2.1±0.1 g), lean mass (21.6±0.2 vs 19.8±0.2 g), as well as kidney weight (491.3±7.69 vs 303.1±5.5 mg) compared to controls. DHT increased fasting glucose (183.2±25.2 vs 141.5±9.9 mg/dL, p<0.05) and impaired OGTT (AUC:34557±6255 vs 23459±1896 mg.min/dL, p<0.05). ARV-110 treatment ameliorated all aforementioned DHT-mediated changes. At the molecular level, DHT-treated mice had higher renal AR (2.5-fold) and MMP9 (1.5-fold) protein expression. DHT upregulated renal TGF-β levels (2-fold, p<0.05), UACR (966.8±160.9 vs 381.5±41.2 ng albumin/mg creatinine), and urinary KIM1 (3.5-fold) and NGAL (6-fold). ARV-110 successfully decreased renal AR protein levels by 70% in DHT mice compared to vehicle-treated DHT. Excitingly, while ARV-110 did not modulate renal TGF-β levels, it completely abolished DHT-mediated increase in renal MMP9 while significantly lowering the levels of urinary renal injury markers UACR, KIM1 and NGAL. Conclusion and significance: Our findings suggest that DHT-mediated kidney hypertrophy, associated ECM remodeling and injury are ameliorated by ARV-110 in DHT-treated mice. AR PROTACs could be a novel therapeutic approach to treat renal dysfunction in PCOS. Supported by NIH P20GM121334. Presentation Date: Sunday, June 18, 2023

Serum Prolactin as an Early Biomarker for Detection of Gestational Diabetes Mellitus-An Experience at CMH Quetta

Objective: To determine the association between elevated serum prolactin and the development of gestational diabetes mellitus in the first trimester.
 Study Design: Prospective longitudinal study.
 Place and Duration of Study: Pathology Department, Combined Military Hospital, Quetta Pakistan, from Apr 2021 to Mar 2022.
 Methodology: One hundred and twenty-one patients had a measurement of serum prolactin in the first trimester, along with an oral glucose tolerance test in the second and third trimester, at the Pathology Department Combined Military Hospital Quetta Pakistan.
 Results: In the study group, serum prolactin levels were raised in 51(42.1%) patients in the first trimester. A total of 44(36.3%) patients with raised serum prolactin had impaired oral glucose tolerance tests. A significant association was found between raised serum prolactin and subsequent development of gestational diabetes mellitus in pregnant females.
 Conclusion: Serum prolactin can be an important biomarker for early diagnosis and monitoring of gestational diabetes mellitus.

Once a week consumption of Western diet over twelve weeks promotes sustained insulin resistance and non-alcoholic fat liver disease in C57BL/6 J mice

The Western diet (high in fat and sucrose) consumption is a highly prevalent feature in the whole world, mainly due to the increasing consumption of ultra-processed foods (UPF), which are cheaper and easier-to-eat, as compared to fresh and highly nutritive meals. Epidemiological studies have associated UPF consumption with development of obesity, non-alcoholic fat liver disease (NAFLD) and insulin resistance. For molecular studies, mice fed with Western diets have been used to characterize signaling pathways involved in these diet-induced pathologies. However, these studies fed mice continuously with the diets, which is not compatible with what occurs in real life, when consumption is occasional. Here, we fed mice once-a-week with a high fat, high sucrose (HFHS) diet and compared these animals with those fed continuously with HFHS diet or with a standard diet. Our results show that after a single day of consuming HFHS, animals presented impaired oral glucose tolerance test (oGTT) as compared to control group. Although this impairment was reversed after 24 h consuming regular diet, repetition of HFHS consumption once-a-week aggravated the picture such as after 12-weeks, oGTT impairment was not reversed after 6 days under control diet. Liver steatosis, inflammation, impaired insulin signaling pathway and endoplasmic reticulum stress are similar comparing animals that consumed HFHS once-a-week with those that continuously consumed HFHS, though weekly-fed animals did not gain as much weight. Therefore, we conclude that regimen of one day HFHS plus 6 days normal diet over 12 weeks is sufficient to induce insulin resistance and NAFLD in mice.

RF10 | PMON249 MicroRNA-21 Ablation Exacerbates Androgen-Mediated White Adipose Tissue Dysfunction and Impaired Insulin Signaling in a Mouse Model of Polycystic Ovary Syndrome

Abstract Background Polycystic ovary syndrome (PCOS) is characterized by oligo- or anovulation, polycystic ovaries, and hyperandrogenism. PCOS is associated with white adipose tissue (WAT) expansion and dysfunction, which has been linked to impaired insulin signaling in a number of metabolic diseases. MicroRNAs dysregulation in WAT is involved in the development of obesity and its associated complications. MicroRNA-21 (miR-21), the most abundant circulatory microRNA in women with PCOS, regulates androgen receptor (AR) expression in prostate cancer however, its role in WAT dysfunction in PCOS, and the associated systemic insulin resistance, is unknown. Using a well-established PCOS model of hyperandrogenemia we aim to test the hypothesis that miR-21 regulates AR expression to modulate androgen-mediated WAT dysfunction and the associated local and systemic insulin resistance. Methods Three-week-old miR-21 knockout (miR-21KO) or wild type (WT) female mice were treated with dihydrotestosterone (DHT, 8 mg/Silastic tube) or vehicle for 90 days (n=6/group). Body composition (EchoMRI) and oral glucose tolerance test (OGTT) were assessed. HOMA-IR index was calculated from fasting serum glucose and insulin levels. Retroperitoneal fat (RPF), a WAT depot that is linked to poor plasma glycemic indices and systemic inflammation, was collected to assess adipocyte size and perform molecular determinations. RPF miR-21 levels, as well as gene and protein expression of AR, and insulin signaling markers (IRS-1/2, insulin receptor-β, PI3K, AKT, Glut4, GSK-α, PTEN) were quantified by RT-qPCR and Western blot. Results In WT mice, DHT increased body weight (25.07 ± 0.52 vs 21.79 ± 0.47 g, p<0.05), fat mass (4.60 ± 0.46 vs 1.98 ± 0.12 g, p<0.05), RPF mass (29.68 ± 3.39 vs. 172.64 ± 32.75 g, p<0.05), RPF miR-21 (2.5-fold) and AR expression (2.1-fold). Adipocyte size analysis revealed a hypertrophic response to DHT, which was associated with impaired OGTT (186.10 ± 5.99 vs 250.70 ± 14.76 mg. min/dL, p<0.05) and downregulation of IRS-1/2, insulin receptor-β, and AKT, indicating impaired insulin signaling. MiR-21 ablation had no effect on DHT-mediated total fat or RPF mass increases, but it exacerbated DHT-mediated AR upregulation, RPF hypertrophy, and insulin resistance as measured by the HOMA-IR index. DHT-mediated reduction in GSK-α, which mediates obesity-induced inflammation, was abolished in miR-21KO mice. Glut4 protein was only downregulated in miR-21KO DHT-treated mice, despite DHT downregulating Glut4 mRNA in both strains. Furthermore, DHT only upregulated PTEN, a negative insulin signaling regulator and known target of miR-21 in miR-21KO mice. Conclusion and significance: These findings imply that WAT miR-21 plays a protective role in PCOS by alleviating androgen-mediated adipose tissue structural and molecular derangements, as well as the associated local and systemic insulin resistance via downregulating AR. Modulation of miR-21 levels in adipose tissue could be a novel therapeutic approach for the treatment of PCOS-related metabolic derangements. Presentation: Saturday, June 11, 2022 1:00 p.m. - 1:05 p.m., Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Polycystic Ovarian Syndrome in Aging Women: An Observational Study.

Background: Polycystic ovarian syndrome (PCOS) in aging women has not been extensively studied, although it is a very common disorder. Most of the research has been conducted on women of the reproductive age group, although PCOS is a disease occurring in all age groups. This study aims to provide an idea of the PCOS pattern prevalent in aging women presenting at a tertiary care center in Eastern India.Methods: This was a prospective, single-center, observational cohort study conducted over three years, from October 2016 to September 2019. A total of 36 patients between 35 and 65 years of age were found to have PCOS according to Rotterdam’s criteria during this period. These patients were prospectively followed up to study their demographic characteristics, symptoms, management, and outcomes.Results: A total of 80% of the women were below 40 years of age, while only 20% were above 40. In total, 70% of the women were overweight or obese. Infertility was seen in 75% of patients below 40 years of age and 28% of patients above 40. Among these, 23 (95%) conceived successfully after proper management. The main presenting complaints were infertility and menstrual irregularities. With aging, diabetes increased from 24%, in below 40, to 28%, in above 40; hypertension increased from 13% to 28%. The occurrence of impaired oral glucose tolerance test (OGTT) and impaired lipid profile increased with age: from 48% to 57% and 13% to 28%, respectively.Conclusion: PCOS in aging women causes considerable morbidity and greatly affects the quality of life, although it is less understood. Further research in this field is very much needed to understand and appropriately manage the problems in aging women, to improve their quality of life.

Glycemic indices at night measured by CGM are predictive for a lower pulmonary function in adults but not in children with cystic fibrosis

IntroductionIn patients with cystic fibrosis (CF), it is still unclear to which extent glucose abnormalities – preceding the diagnosis of cystic fibrosis related diabetes (CFRD) – are associated with pulmonary and nutritional outcome parameters. This study related circadian glycemic patterns to clinical outcomes in a group of CF patients not previously diagnosed with diabetes. MethodsContinuous glucose monitoring (CGM) readings (7 days) of 47 CF patients (26 children, 21 adults) with an impaired oral glucose tolerance test (OGTT) (n = 25) and/or increased Hb1Ac (> 5.5%) were analyzed. Biometric, pulmonary function and clinical parameters were retrospectively collected over a period of 1 year before (T-1) and 1 year after (T + 1) CGM (T0). Results96% (45/47) of CGM readings showed glucose values > 140 mg/dL ≥ 4.5% of the time and at least one ≥ 200 mg/dL. In the pediatric cohort, no significant associations were found between CGM parameters and pulmonary and nutritional outcome parameters. In the adult cohort, an area under the curve (AUC) > 140 mg/dL and%-time > 140 mg/dL during the night were associated with a lower forced expiratory volume in 1 s (FEV1)% predicted (pp) at time of evaluation but not with change in FEV1pp. ConclusionThis is the first study reporting the circadian glycemic pattern in children and adults at risk for CFRD. In the adult cohort an association between detection of abnormal glucose exposure and a lower FEV1pp was found. Our results support continued screening for glucose intolerance in patients with CF.

The impact of prediabetes in the first trimester of pregnancy on the incidence of gestational diabetes

Introduction: Prediabetes (PD) outside of pregnancy is a condition reflecting a significant loss of glucose tolerance, characterized in the laboratory by fasting glucose between 100 and 125 mg/dL or 2 hour glucose in the 75 g Oral Glucose Tolerance Test (OGTT) between 140 and 199 mg/dL or by HbA1c levels between 5.7%-6.4% and is associated with an increased risk of developing type 2 diabetes. The development of gestational diabetes mellitus (GDM) during pregnancy, characterized by a fasting blood glucose between 92 and 125 mg/dL or an impaired OGTT value of 75 g between 24th and 28th weeks, is another condition that increases the risk of perinatal complications such as preeclampsia, cesarean section rate, macrosomia, and also the development of type 2 diabetes after delivery. Determining the prevalence of women diagnosed with prediabetes at the beginning of prenatal care and its association with the diagnosis of GDM is therefore of great importance for clinical, gynecological and obstetric care. Objective: the aim of the study is to determine the prevalence and association of prediabetes in the first trimester of pregnancy with the development of gestational diabetes
 Methodology: It was carried out in the Clinical Hospital of Botucatu Medical School. Retrospective cohort study involving eligible pregnant women who underwent fasting glucose test (FG) and glycated hemoglobin (HbA1c) test up to the 14 th week of gestation without previous diagnosis of diabetes mellitus, and who underwent 75 g oral glucose tolerance test (between the 24th and 28th week). We excluded cases in which FG was between 92 mg/dL and 125 mg/dL, which were considered GDM, and cases with FG ≥ 126 mg/dL and/or HbA1c ≥ 6.5 % (overt diabetes). One thousand electronic medical records from between 2013 and 2021 were reviewed and a database was created. The partial results were presented as percentage and 95 Confidence Interval (95% CI). The association between HbA1c and GDM was presented as relative risk and 95% CI. Results: Among pregnant women with normal fasting glucose levels and HbA1c <5.7%, the incidence of GDM, diagnosed by OGTT, was 8.5% (6.4-11.3), while in the group with PD was 14.3% (5.8%-29.8%). The relative risk (RR) for developing GDM among the PD group was 1.7 (0.7-3.9). Discussion and conclusion: The association between prediabetes and gestational diabetes was not confirmed. However, these results are preliminary, and we need to reach the sample size calculated before starting the study to better understand these results. Furthermore, nonpregnant women with prediabetes are monitored and eventually treated, therefore, further studies are needed to evaluate the benefit of treating these pregnant women as recommended for the general population

MicroRNA-21 Modulates White Adipose Tissue Browning and Altered Thermogenesis in a Mouse Model of Polycystic Ovary Syndrome

Background and Aim: Polycystic ovary syndrome (PCOS) is associated with obesity, and white adipose tissue (WAT) and brown adipose tissue (BAT) dysregulation. However, the molecular mechanisms that mediate WAT and BAT derangements in PCOS are poorly understood. Subcutaneous (SC) WAT (SC-WAT) can transition to a beige/brite adipose tissue phenotype (browning) under altered thermogenic conditions. MicroRNAs play critical functions in brown adipocyte differentiation and maintenance. We aim to study the role of microRNA-21 (miR-21) in androgen-mediated browning and beiging derangements in both SC-WAT and BAT. Methods: Three week-old miR-21 knockout (miR21KO) or wild type (WT) female mice were treated with dihydrotestosterone (DHT, 8 mg/silastic tube) or vehicle for 90 days (n=12/grp). Body composition was measured by EchoMRI. Energy expenditure (EE), oxygen consumption (VO2), and carbon dioxide production (VCO2) were measured by indirect calorimetry. Glucose homeostasis was measured by oral glucose tolerance test (OGTT). HOMA-IR index was calculated from fasting serum glucose and insulin levels. Gene expression for browning (UCP1, Cox7a1, Elov3, Dio2 and Cidea) and beiging (Hspb7 and Txb1) markers was quantified by RT-qPCR in SC-WAT and BAT. Results: DHT increased body weight (25.07 ± 0.52 vs 21.79 ± 0.47 g, p<0.05) and fat mass (4.60 ± 0.46 vs 1.98 ± 0.12 g, p<0.05), impaired OGTT (186.10 ± 5.99 vs 250.70 ± 14.76 mg.min/dL, p<0.05), and did not significantly change EE, VO2 or VCO2 in WT mice. All browning markers were downregulated by DHT in SC-WAT; however, only iodothyronine deiodinase 2 (Dio2) downregulation reached significance in both SC-WAT and BAT (by 53 and 40%, respectively) compared with the vehicle-treated mice. Beiging markers were significantly upregulated in SC-WAT and did not change in BAT. DHT-treated miR21KO mice showed attenuated DHT-mediated increase in body weight (23.84 ± 0.99 vs 25.07 ± 0.52 g, p<0.05) compared with WT mice. MiR-21 ablation did not modify DHT-mediated increase in fat mass or OGTT but worsened insulin resistance as calculated by the HOMA-IR index. Additionally, DHT-treated miR21KO mice showed a trend to reduced EE, VO2 and VCO2 values compared with DHT-treated WT. Gene expression analysis showed an exacerbation in DHT-mediated reduction in browning markers expression in the SC-WAT. Additionally, the induction in the adaptive beiging response was abolished in SC-WAT. Conclusion and Significance: These findings suggest that adipose tissue miR-21 may have a protective role in PCOS and ameliorate the DHT-mediated decrease in energy expenditure. Adipose tissue-specific modulation of miR-21 levels could be a novel therapeutic approach for the treatment of PCOS-associated metabolic derangements. (Supported by NIH grants NIGMS P20GM121334 to LLYC and DGR, NIDDK R21DK113500 to DGR, NIGMS P20GM104357 and NHLBI P01HL51971).

Effect of Hydrolyzed Bird's Nest on β-Cell Function and Insulin Signaling in Type 2 Diabetic Mice.

Type 2 diabetes mellitus is characterized by both resistance to the action of insulin and defects in insulin secretion. Bird’s nest, which is derived from the saliva of swiftlets are well known to possess multiple health benefits dating back to Imperial China. However, it’s effect on diabetes mellitus and influence on the actions of insulin action remains to be investigated. In the present study, the effect of standardized aqueous extract of hydrolyzed edible bird nest (HBN) on metabolic characteristics and insulin signaling pathway in pancreas, liver and skeletal muscle of db/db, a type 2 diabetic mice model was investigated. Male db/db diabetic and its euglycemic control, C57BL/6J mice were administered HBN (75 and 150 mg/kg) or glibenclamide (1 mg/kg) orally for 28 days. Metabolic parameters were evaluated by measuring fasting blood glucose, serum insulin and oral glucose tolerance test (OGTT). Insulin signaling and activation of inflammatory pathways in liver, adipose, pancreas and muscle tissue were evaluated by Western blotting and immunohistochemistry. Pro-inflammatory cytokines were measured in the serum at the end of the treatment. The results showed that db/db mice treated with HBN significantly reversed the elevated fasting blood glucose, serum insulin, serum pro-inflammatory cytokines levels and the impaired OGTT without affecting the body weight of the mice in all groups. Furthermore, HBN treatment significantly ameliorated pathological changes and increased the protein expression of insulin, and glucose transporters in the pancreatic islets (GLUT-2), liver and skeletal muscle (GLUT-4). Likewise, the Western blots analysis denotes improved insulin signaling and antioxidant enzyme, decreased reactive oxygen species producing enzymes and inflammatory molecules in the liver and adipose tissues of HBN treated diabetic mice. These results suggest that HBN improves β-cell function and insulin signaling by attenuation of oxidative stress mediated chronic inflammation in the type 2 diabetic mice.

Targeting inhibition of CCR5 on improving obesity-associated insulin resistance and impairment of pancreatic insulin secretion in high fat-fed rodent models

Obesity is closely linked with type 2 diabetes and the effective therapies on obesity-associated diabetes are under development. The aim of this study was undertaken to investigate whether the inhibition of the augmented CCR5-mediated signaling could be a common target for treatment of obesity-associated insulin resistance and impairment of pancreatic insulin secretion in high-fat diet (HFD) fed rats and CCR5 knockout mice and also in isolated islets and RIN-m5F cells. Conducted with SD rats, HFD-induced body weight gain was significantly decreased in those combined with Maraviroc treatment, but food intake remained similar compared to control. Maraviroc also significantly improved the impaired oral glucose tolerance test (OGTT). As compared with wild-type mice, CCR5 deletion significantly attenuated the HFD-induced increases in glucose area under curve of OGTT and the value of HOMA-IR as well as plasma lipid profile. It also reversed the HFD-suppressed gene expressions of GLUT4 and IRS-1 in adipose tissue. On the other hand, the HFD-associated islet macrophage and T-cell infiltration were significantly decreased in CCR5 KO mice. H2O2 significantly suppressed glucose-stimulated insulin secretion (GSIS) is isolated islets, which were significantly reversed in those cotreated with CCR5 mAb. H2O2 failed to change GSIS in those of CCR5 KO mice. The palmitate-induced reactive oxygen species production was significantly decreased in those cotreated with CCR5 antagonist in RIN-m5F cells. Collectively, it is suggested that targeting inhibition of the CCR5 mediated inflammatory pathway could not only improve obesity-associated insulin resistance but also directly alleviate pancreatic β-cell dysfunction.

Maternal Metabolic Alterations in Monosodium Glutamate Fed Rats during Gestation and Lactation Period

Background: Monosodium glutamate (MSG), an extensively used food additive, is claimed to cause many health problems, its use is still under debate. Aim: to explore the metabolic and hypertensive effect of MSG dietary administration in pregnant and lactating female rats, and its possible underlying mechanism. Methods: 16 adult female albino Wister rats were divided into 2 groups, control group; were fed standard rat chow, and MSG group; were fed 2% MSG supplemented rat chow throughout gestation and lactation period. Oral glucose tolerance test (OGTT) and arterial blood pressure (ABP) were measured at mid and late gestation, mid and late lactation periods. At mid lactation period, a retroorbital samples were analyzed for fasting glucose, lipid profile; triglycerides (TGs), total cholesterol (TC) and low density lipoprotein-cholesterol (LDL-C), hormonal assays; insulin, glucagon, prolactin, corticosterone and ACTH, as well as pancreatic lipase, amylase and malondialdehyde (MDA). By the end of lactation, adipose tissue was assessed for glucose transporter-4 (GLUT-4) and hormone sensitive lipase (HSL) relative expression. Results: MSG rats showed elevated ABP during pregnancy and lactation, with impaired OGTT at mid and late lactation period. Fasting serum glucose, TGs, TC, LDL-C, glucagon, corticosterone, pancreatic lipase and amylase, MDA as well as adipose tissue HSL expression were significantly elevated, while, fasting serum insulin, ACTH and adipose tissue GLUT-4 expression were significantly declined in MSG fed rats as compared to control group. Conclusion: MSG feeding during pregnancy and lactation induced hypertension and metabolic alterations that could be due to pancreatic and adrenal affection secondary oxidative stress.

Transcription Factor HMG Box-Containing Protein 1 (HBP1) Regulates Glycemic Homeostasis Through Modulation of Gluconeogenesis

Abstract Objectives Glycemic dysregulation is one of the major metabolic disorders, which has long been a significant public health issue. The liver plays a pivotal role in the maintenance of blood glucose homeostasis. Previous studies indicate that upon refeeding a high carbohydrate diet, the expression of transcription factor HMG box-containing protein 1 (HBP1) was elevated in mice livers, suggesting a role of HBP1 in carbohydrate metabolism. Therefore, the objective of the current study was to understand the molecular mechanisms through which HBP1 regulates glucose generation in the liver. Methods Both in vivo HBP1 knockdown mice and in vitro HepG2 cell line were employed as the experimental models. Results First, we observed that overnight fasting led to increased PEPCK but decreased HBP1 expression in mice livers, and subsequent addition of insulin reversed the expression pattern. More importantly, HBP1 knockout (KO) mice displayed a significantly higher blood glucose level (173 mg/dL) than that of the controls (98 mg/dL). Also, HBP1 KO led to impaired OGTT (oral glucose tolerance test) and ITT (insulin tolerance test). These data suggest that HBP1 might have a role in the regulation of gluconeogenesis in the liver. To unveil the molecular mechanism by which HBP1 regulates glucose homeostasis, we examined its role in gluconeogenesis. Administration of gluconeogenic stimulators glucagon (100 nM) and cAMP (0.5 ng/mL) resulted in increased expression of Phosphoenolpyruvate carboxykinase (PEPCK; encoded by the PCK1 gene), a key enzyme in gluconeogenesis, but decreased HBP1 expression in HepG2 cells. Last, HBP1 siRNA-mediated mRNA disruption led to elevated PEPCK expression, whereas ectopic expression of HBP1 (pcDNA3-HBP1-Flag) significantly suppressed it. Conclusions In conclusion, our data indicate that HBP1 might negatively regulate glucose production and support HBP1 as a novel biological regulator of blood glucose homeostasis. Funding Sources This work was supported by the grant to MOST 108–2320-B-039–051-MY3 C-Y Huang.

Effectiveness of Bariatric Surgery-Induced Weight Loss on Infertility Among PCOS and Non-PCOS Women: Experience of a Maternity Hospital and In Vitro Fertilization (IVF) Center in India

Obesity, mostly the abdominal deposition of fat, is prevalent with polycystic ovarian syndrome (PCOS) influencing fertility. Evidence suggests a significant surge in the number of women who became pregnant after bariatric surgery (BS). The objective of this study is to evaluate the role of bariatric surgery-induced weight loss prior to pregnancy on various parameters, with special focus on infertility among women with and without PCOS. A single-center retrospective cohort study, comprising 28 women {PCOS (n = 17) and without PCOS (n = 11)} with a history of infertility, who underwent bariatric surgery for morbid obesity. Retrospective analysis was performed for pregnancy rate, weight loss (in terms of body mass index), anti-mullerian hormone (AMH) levels, oral glucose tolerance test (OGTT) value, and serum insulin levels. The mean of total pregnancy rate was 64.28%. At 1-year follow-up, the mean BMI among all the study participants decreased from 49.11 pre-surgery to 29.33 post-surgery. Similarly, the prevalence of impaired OGTT reduced from 42.85 to 3.57% among the study groups. The AMH levels were within the normal range for both the groups after surgery, but were not statistically significant. The study showed the positive role of bariatric surgery on overall health in both groups. Also, bariatric surgery was favorable on the reproductive health of women as well as in managing other metabolic co-morbidities like impaired OGTT and serum insulin levels in PCOS as well as non-PCOS women.

Depot‐Specific Response of White Adipose Tissue to MicroRNA‐21 Ablation in Polycystic Ovary Syndrome

Background and aimPolycystic ovary syndrome (PCOS) is associated with obesity and insulin resistance. However, the molecular mechanisms that mediate this phenotype remain elusive. MicroRNA‐21 (miR‐21) is upregulated in PCOS and may be involved in the pathogenesis of the disease. We aim to study the role of miR‐21 in the deleterious metabolic effects associated with PCOS.MethodsThree week‐old miR‐21 knockout (miR21KO) or wild type (WT) female mice were treated with DHT (8 mg/silastic tube) or vehicle for 90 days (n=12/grp). Body composition was measured by EchoMRI. Energy expenditure (EE), oxygen consumption (VO2), and carbon dioxide production (VCO2) were measured by indirect calorimetry. Glucose homeostasis was measured by oral glucose tolerance test (OGTT). Gene expression was quantified by RT‐qPCR in subcutaneous (SC), retroperitoneal (RP) and mesenteric (M) white adipose tissue (WAT). Adipocyte size was evaluated by histological analysis.ResultsDHT increased body weight (25.07 ± 0.52 vs 21.79 ± 0.47 g, p&lt;0.05), fat mass (4.60 ± 0.46 vs 1.98 ± 0.12 g, p&lt;0.05) and adipocyte size (944 ±12 vs 513 ± 2 μm2, p&lt;0.05), impaired OGTT (29.70 ± 2.21 vs 22.33 ± 1.06 AUC, p&lt;0.05), and didn’t significantly change EE, VO2 or VCO2. Insulin signaling markers Irs1 and Slc2a4 expression was downregulated by DHT in all 3 WATs while Akt2 expression was upregulated. Adipogenesis markers Pparg and Adipoq were upregulated by DHT exclusively in the SC WAT. Inflammation marker Ccl2 was upregulated in all three WATs while Ikbkb was only upregulated in M WAT. Extracellular matrix (ECM) remodeling makers Mmp9 and Timp1 were upregulated by DHT only in RP and M WATs. Immune cells infiltration markers CD14, CD68, Itgam, Itgax, and CD86 were upregulated by DHT only in RP and M WATs.DHT‐treated miR21KO mice showed attenuated increase in body weight (23.84 ± 0.99 vs 25.07 ± 0.52 g, p&lt;0.05) with no change in fat mass or OGTT. Additionally, DHT‐treated miR21KO mice showed a trend to reduced EE, VO2 and VCO2 compared with DHT‐treated WT. Gene expression data showed lower expression of the insulin signaling marker Akt2, higher expression of ECM remodeling markers (Mmp9 and Timp1) and an exacerbated inflammatory and immune responses in M (Il‐18 and CD14) and SC (Il‐18 and CD86) WATs. Moreover, DHT‐treated miR21KO mice had exacerbated adipocyte hypertrophy compared with DHT‐treated WT controls (1,408 ± 16 vs 944 ±12 μm2, p&lt;0.05).Conclusion and significanceThese findings suggest that adipose tissue miR‐21 may have a protective role in PCOS and ameliorate some of the DHT‐mediated molecular changes observed in WAT in a fat depot‐specific manner. WAT‐specific modulation of miR‐21 levels could be a novel therapeutic approach for PCOS treatment.Support or Funding InformationSupported by National Institutes of Health grants P20 GM‐121334 (DGR and LLYC) and R21 DK‐113500 (DGR).

ETHANOLIC EXTRACT OF TUBTIM-CHUMPHAE RICE BRAN DECREASES INSULIN RESISTANCE AND INTRAHEPATIC FAT ACCUMULATION IN HIGH-FAT-HIGH-FRUCTOSE DIET FED RATS

Objective: Tubtim-chumphae rice is hybrid Thai rice with a red pericarp. This study was aimed to investigate the effect of Tubtim-chumphae rice bran on insulin resistance and intrahepatic fat accumulation in high-fat-high-fructose diet (HFFD) fed rats.Methods: Ethanolic extract of rice bran (ERB) was prepared using a 50% ethanol-water. Male Sprague-Dawley rats were fed HFFD (40% lard, 20% fructose) for 10 weeks, followed by concomitant administrations of distilled water or ERB at 250 or 500 mg/kg/day or pioglitazone at 10 mg/kg/day for a further 4 weeks in treated groups. Normal control rats were fed normal chow and distilled water. At the end of all treatments, fasting blood glucose (FBG), an oral glucose tolerance test (OGTT), serum insulin levels, lipid profiles, and liver fat contents were measured. Liver histological and peroxisome proliferator-activated receptor-α (PPAR-α) gene expression examinations were performed.Results: At week 14, control HFFD rats had significantly (p&lt;0.05) higher FBG, low-density lipoprotein cholesterol, triglycerides, and insulin secretions together with impaired OGTT as compared to normal control rats. These parameters indicated an insulin resistant and dyslipidemic condition in HFFD rats. ERB 250 and 500 mg/kg or pioglitazone 10 mg/kg significantly ameliorated all of these changes. HFFD also caused a significant increase in fat accumulation and a decrease in PPAR-α gene expression in the livers which were significantly decreased by ERB.Conclusions: ERB decreases insulin resistance and intrahepatic fat accumulation possibly through increasing PPAR-α gene expression in HFFD rats. ERB might possibly be a neutraceutical for the metabolic syndrome patients.1. Gauthier MS, Favier R, Lavoie JM. Time course of the development of non-alcoholic hepatic steatosis in response to high-fat diet-induced obesity in rats. Br J Nutr 2006;95:273-81.2. Roberts CK, Hevener AL, Barnard RJ. Metabolic syndrome and insulin resistance: Underlying causes and modification by exercise training. Compr Physiol 2013;3:1-58.3. Grundy SM. Metabolic syndrome update. Trends Cardiovasc Med 2016;26:364-73.4. Fouret G, Gaillet S, Lecomte J, Bonafos B, Djohan F, Barea B, et al. 20-week follow-up of hepatic steatosis installation and liver mitochondrial structure and activity and their interrelation in rats fed a high-fat-high-fructose diet. Br J Nutr 2018;119:368-80.5. Dekker MJ, Su Q, Baker C, Rutledge AC, Adeli K. Fructose: A highly lipogenic nutrient implicated in insulin resistance, hepatic steatosis, and the metabolic syndrome. Am J Physiol Endocrinol Metab 2010;299:E685-94.6. Vichit W, Saewan N. Antioxidant activities and cytotoxicity of thai pigmented rice. Int J Pharm Pharm Sci 2015;7:329-34.7. Settharaksa S, Madaka F, Charkree K, Charoenchai L. The study of anti-inflammatory and antioxidant activity in cold press rice bran oil from rice in Thailand. Int J Pharm Pharm Sci 2014;6:428-31.8. Sukrasno S, Tuty S, Fidrianny I. Antioxidant evaluation and phytochemical content of various rice bran extracts of three varieties rice from Semarang, central Java, Indonesia. Asian J Pharm Clin Res 2017;10:377-82.9. Sabir A, Rafi M, Darusman LK. Discrimination of red and white rice bran from indonesia using HPLC fingerprint analysis combined with chemometrics. Food Chem 2017;221:1717-22.10. Niu Y, Gao B, Slavin M, Zhang X, Yang F, Bao J, et al. Phytochemical compositions, and antioxidant and anti-inflammatory properties of twenty-two red rice samples grown in Zhejiang. LWT Food Sci Technol 2013;54:521-7.11. Boonloh K, Kukongviriyapan V, Kongyingyoes B, Kukongviriyapan U, Thawornchinsombut S, Pannangpetch P, et al. Rice bran protein hydrolysates improve insulin resistance and decrease pro-inflammatory cytokine gene expression in rats fed a high carbohydrate-high fat diet. Nutrients 2015;7:6313-29.12. Peñarrieta JM, Alvarado JA, Akesson B, Bergenståhl B. Total antioxidant capacity and content of flavonoids and other phenolic compounds in canihua (Chenopodium pallidicaule): An andean pseudocereal. Mol Nutr Food Res 2008;52:708-17.13. Mungkhunthod S, Senggunprai L, Tangsucharit P, Sripui J, Kukongviriyapan U, Pannangpetch P. Antidesma thwaitesianum pomace extract improves insulin sensitivity via upregulation of PPAR-γ in high fat diet/streptozotocin-induced Type 2 diabetic rats. Asia Pac J Sci Technol 2016;21:63-76.14. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC, et al. Homeostasis model assessment: Insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 1985;28:412-9.15. Naowaboot J, Wannasiri S. Anti-lipogenic effect of Senna alata leaf extract in high-fat diet-induced obese mice. Asian Pac J Trop Biomed 2016;6:232-8.16. Couturier K, Qin B, Batandier C, Awada M, Hininger-Favier I, Canini F, et al. Cinnamon increases liver glycogen in an animal model of insulin

ETHANOLIC EXTRACT OF TUBTIM-CHUMPHAE RICE BRAN DECREASES INSULIN RESISTANCE AND INTRAHEPATIC FAT ACCUMULATION IN HIGH-FAT-HIGH-FRUCTOSE DIET FED RATS

Objective: Tubtim-chumphae rice is hybrid Thai rice with a red pericarp. This study was aimed to investigate the effect of Tubtim-chumphae rice bran on insulin resistance and intrahepatic fat accumulation in high-fat-high-fructose diet (HFFD) fed rats.Methods: Ethanolic extract of rice bran (ERB) was prepared using a 50% ethanol-water. Male Sprague-Dawley rats were fed HFFD (40% lard, 20% fructose) for 10 weeks, followed by concomitant administrations of distilled water or ERB at 250 or 500 mg/kg/day or pioglitazone at 10 mg/kg/day for a further 4 weeks in treated groups. Normal control rats were fed normal chow and distilled water. At the end of all treatments, fasting blood glucose (FBG), an oral glucose tolerance test (OGTT), serum insulin levels, lipid profiles, and liver fat contents were measured. Liver histological and peroxisome proliferator-activated receptor-α (PPAR-α) gene expression examinations were performed.Results: At week 14, control HFFD rats had significantly (p&lt;0.05) higher FBG, low-density lipoprotein cholesterol, triglycerides, and insulin secretions together with impaired OGTT as compared to normal control rats. These parameters indicated an insulin resistant and dyslipidemic condition in HFFD rats. ERB 250 and 500 mg/kg or pioglitazone 10 mg/kg significantly ameliorated all of these changes. HFFD also caused a significant increase in fat accumulation and a decrease in PPAR-α gene expression in the livers which were significantly decreased by ERB.Conclusions: ERB decreases insulin resistance and intrahepatic fat accumulation possibly through increasing PPAR-α gene expression in HFFD rats. ERB might possibly be a neutraceutical for the metabolic syndrome patients.1. Gauthier MS, Favier R, Lavoie JM. Time course of the development of non-alcoholic hepatic steatosis in response to high-fat diet-induced obesity in rats. Br J Nutr 2006;95:273-81.2. Roberts CK, Hevener AL, Barnard RJ. Metabolic syndrome and insulin resistance: Underlying causes and modification by exercise training. Compr Physiol 2013;3:1-58.3. Grundy SM. Metabolic syndrome update. Trends Cardiovasc Med 2016;26:364-73.4. Fouret G, Gaillet S, Lecomte J, Bonafos B, Djohan F, Barea B, et al. 20-week follow-up of hepatic steatosis installation and liver mitochondrial structure and activity and their interrelation in rats fed a high-fat-high-fructose diet. Br J Nutr 2018;119:368-80.5. Dekker MJ, Su Q, Baker C, Rutledge AC, Adeli K. Fructose: A highly lipogenic nutrient implicated in insulin resistance, hepatic steatosis, and the metabolic syndrome. Am J Physiol Endocrinol Metab 2010;299:E685-94.6. Vichit W, Saewan N. Antioxidant activities and cytotoxicity of thai pigmented rice. Int J Pharm Pharm Sci 2015;7:329-34.7. Settharaksa S, Madaka F, Charkree K, Charoenchai L. The study of anti-inflammatory and antioxidant activity in cold press rice bran oil from rice in Thailand. Int J Pharm Pharm Sci 2014;6:428-31.8. Sukrasno S, Tuty S, Fidrianny I. Antioxidant evaluation and phytochemical content of various rice bran extracts of three varieties rice from Semarang, central Java, Indonesia. Asian J Pharm Clin Res 2017;10:377-82.9. Sabir A, Rafi M, Darusman LK. Discrimination of red and white rice bran from indonesia using HPLC fingerprint analysis combined with chemometrics. Food Chem 2017;221:1717-22.10. Niu Y, Gao B, Slavin M, Zhang X, Yang F, Bao J, et al. Phytochemical compositions, and antioxidant and anti-inflammatory properties of twenty-two red rice samples grown in Zhejiang. LWT Food Sci Technol 2013;54:521-7.11. Boonloh K, Kukongviriyapan V, Kongyingyoes B, Kukongviriyapan U, Thawornchinsombut S, Pannangpetch P, et al. Rice bran protein hydrolysates improve insulin resistance and decrease pro-inflammatory cytokine gene expression in rats fed a high carbohydrate-high fat diet. Nutrients 2015;7:6313-29.12. Peñarrieta JM, Alvarado JA, Akesson B, Bergenståhl B. Total antioxidant capacity and content of flavonoids and other phenolic compounds in canihua (Chenopodium pallidicaule): An andean pseudocereal. Mol Nutr Food Res 2008;52:708-17.13. Mungkhunthod S, Senggunprai L, Tangsucharit P, Sripui J, Kukongviriyapan U, Pannangpetch P. Antidesma thwaitesianum pomace extract improves insulin sensitivity via upregulation of PPAR-γ in high fat diet/streptozotocin-induced Type 2 diabetic rats. Asia Pac J Sci Technol 2016;21:63-76.14. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC, et al. Homeostasis model assessment: Insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 1985;28:412-9.15. Naowaboot J, Wannasiri S. Anti-lipogenic effect of Senna alata leaf extract in high-fat diet-induced obese mice. Asian Pac J Trop Biomed 2016;6:232-8.16. Couturier K, Qin B, Batandier C, Awada M, Hininger-Favier I, Canini F, et al. Cinnamon increases liver glycogen in an animal model of insulin

Dipeptidyl peptidase-4 inhibition protects the liver of insulin-resistant female rats against triglyceride accumulation by suppressing uric acid

Dipeptidyl peptidase-4 (DPP-4) inhibition has been shown to exert beneficial effects against insulin resistance (IR) and type 2 diabetes. Combined oral contraceptive (COC) treatment is associated with impaired glucose and lipid metabolism but the mechanisms are elusive. We therefore, hypothesized that DPP-4 inhibition ameliorates COC-induced glucose dysregulation and hepatic triglyceride (TG) accumulation through adenosine deaminase (ADA) /xanthine oxidase (XO) /uric acid-dependent pathway. Female Wistar rats received (po) vehicle and COC (1.0 μg ethinylestradiol plus 5.0 μg levonorgestrel; po) with or without DPP-4 inhibitor (sitagliptin; 100 mg/kg; po) for 8 weeks (n = 6/group). Glucose dysmetabolism was assessed by elevated fasting blood glucose, impaired oral glucose tolerance test and homeostatic model assessment of IR. Treatment with COC led to increased plasma fasting glucose, triglyceride-glucose index, 1-h postload glucose response, insulin, free fatty acid, IR and impaired glucose tolerance. COC treatment also resulted in increased plasma and hepatic TG, TG/HDL-cholesterol ratio, malondialdehyde, uric acid (plasma; 25.2 ± 0.6 mg/dl; hepatic 128.9 ± 8.0 mg/100 mg tissue), lactate dehydrogenase, DPP-4, ADA and XO (plasma;10.5 ± 1.1 U/L; hepatic 21.2 ± 1.4 U/g protein) activities. Likewise, COC led to reduction in nitric oxide level. However, DPP-4 inhibition significantly ameliorated these alterations induced by COC treatment through suppression of uric acid (plasma; 15.1 ± 1.0 mg/dl, hepatic; 75.6 ± 5.0 mg/100 mg tissue), XO (plasma; 4.1 ± 0.9 U/L, hepatic; 8.7 ± 0.4 U/g protein), ADA and DPP-4 activities suggesting their involvement in glucose dysregulation and hepatic TG accumulation induced by COC treatment. Therefore, DPP-4 inhibition would impact positively on cardiometabolic disorders, at least in part, through XO, ADA and uric acid suppression.

A4690 Sodium acetate and mineralocorticoid receptor blockade improve gestational androgen excess-induced glucose dysmetabolism in rats by suppressing adenosine deaminase/xanthine oxidase pathway

Objectives: The effects of sodium acetate and mineralocorticoid receptor (MR) on glucose dysmetabolism and poor fetal outcome induced by gestational androgen excess is not yet well elucidated. We tested the hypothesis that blockade of MR or treatment with sodium acetate will prevent glucose dysmetabolism induced by late gestational androgen excess through suppression of adenosine deaminase (ADA)/xanthine oxidase (XO) pathway. Methods: Thirty-six pregnant Wistar rats were treated (sc) with olive oil, testosterone propionate (0.5 mg/kg) singly or in combination with sodium acetate (200 mg/kg; po), androgen receptor (AR) blocker (flutamide; 7.5 mg/kg; po), non-selective MR blocker (spironolactone 0.25 mg/kg; po) or selective MR blocker (eplerenone; 0.5 mg/kg; po) between gestational days 14 and 19. Glucose dysmetabolism was accessed by elevated fasting blood glucose, impaired oral glucose tolerance test and homeostatic model assessment of insulin resistance. Results: The results showed that late gestational androgen excess led to glucose dysregulation, increased plasma testosterone, plasma and cardiac ADA and XO activities. Conversely, gestational androgen excess resulted in reduced plasma and cardiac anti-oxidant defenses (glutathione peroxidase, reduced glutathione/glutathione disulphide ratio, glucose-6-phosphate dehydrogenase and adenosine). However, all these effects were ameliorated by AR blockade, MR blockade or sodium acetate comparatively. Conclusion: The study demonstrates that improvement of glucose dysregulation induced by late gestational androgen excess by AR blockade, MR blockade or sodium acetate occurs by improvement of anti-ox

A4800 Dipeptidyl peptidase-4 inhibition prevents increased hepatic triglyceride content in insulin-resistant female rats

Objectives: Dipeptidyl peptidase-4 (DPP-4) inhibition has been show to exert beneficial effects against insulin resistance (IR) and fatty liver disease. Combined oral contraceptive (COC) treatment has been shown to be associated with glucose deregulation and increased hepatic triglyceride (TG) levels, but the mechanisms are elusive. We therefore, hypothesized that DPP-4 inhibition (DPP-4i) will ameliorate COC-induced glucose dysregulation and hepatic TG accumulation through adenosine deaminase (ADA) /xanthine oxidase (XO) /uric acid-dependent pathway. Methods: Female Wistar rats received (po) vehicle and COC (1.0 μg ethinylestradiol plus 5.0 μg levonorgestrel; po) with or without DPP-4 inhibitor (sitagliptin; 100 mg/kg; po) for 8 weeks (n = 6/group). Glucose dysmetabolism was accessed by elevated fasting blood glucose, impaired oral glucose tolerance test and homeostatic model assessment of insulin resistance. Results: Treatment with COC led to increased plasma fasting glucose, triglyceride-glucose index, 1-h postload glucose response, insulin, free fatty acid, IR and impaired glucose tolerance. COC treatment also resulted in increased plasma and hepatic triglycerides (TG), TG/HDL-cholesterol ratio, malondialdehyde, uric acid, lactate dehydrogenase, DPP-4, ADA and XO activities. On the other hand, COC led to reduction in nitric oxide level. However, DPP-4i significantly ameliorated these alterations induced by COC treatment except plasma and hepatic XO activity. Conclusion: This demonstrates that increased uric acid, DPP-4 and ADA activities are involved in glucose dysregulation and hepatic TG accumulation induced by COC treatment. Therefore, DPP-4i would impact positively on cardiometabolic disorders, at least in part, through ADA and uric acid suppression.

Risk Factors for Gestational Diabetes Mellitus: A Case-Control Study.

Background: The underlying causes of gestational diabetes mellitus (GDM) are important because they are effective for the diagnosis and prevention of this condition. The aim of this study was to identify the risk factors for GDM and the possible etiological agents. Materials and Methods: This case-control study was conducted with 100 women with GDM and 100 healthy pregnant women at a tertiary care hospital, affiliated to Babol University. GDM was identified by impaired oral glucose tolerance test based on the Carpenter and Coustan criteria. Healthy women were randomly selected from the prenatal health care clinic of the same center and were matched to gestational age of 24 to 28 weeks. Descriptive and inferential statistics were used for data analysis via the SPSS software. Results: After adjusting variables, multivariate analysis identified 4 significant risk factors of GDM, including prepregnancy low physical activity (odds ratio [OR] = 2.85, 95% confidence interval [CI]= 0.97-8.34), advanced maternal age (OR = 1.24, 95% CI = 1.13-1.35), body mass index ⩾ 30 kg/m2 (OR = 1.10, 95% CI = 1.02-1.20), and family history of diabetes mellitus (OR = 5.62, 95% CI = 2.26-13.96). Conclusion: We observed significant associations between low prepregnancy physical activity and obesity with GDM risk. Thus the finding of this study can help devise strategies for the prevention of GDM.