Hepatic Gluconeogenesis Research Articles

Insulin resistance and iatrogenic hyperinsulinemia in type 1 diabetes - norm or complication? Role of metformin in type 1 diabetes mellitus

There is a rising trend of overweight and obesity among individuals with type 1 diabetes. This is often associated with decreased insulin sensitivity, increased insulin dose requirements and poor glycemic control. Insulin resistance is a state of diminished effect of insulin on target tissues, despite normal or elevated serum insulin levels. Metformin as an adjunct to insulin in treating overweight or also not specific image of insulin resistance (without overweight or hormonal disorders) adults with type 1 diabetes is increasingly used. The main goal of this off-label type 1 diabetes treatment method is to improve insulin signaling leading to a subsequent increase in glucose uptake by skeletal myocytes, inhibition of hepatic gluconeogenesis and, as a result, a reduction in the need for a daily dose of insulin. The use of metformin as an addition to insulin may increase the chance of achieving target glucose control and reduce the anabolic effect of this hormone. Metformin can improve insulin sensitivity, glycemia and modulate cardiovascular disease risk factors or nonalcoholic fatty liver disease. In the vast majority of clinical trials conducted with metformin in type 1 diabetes, the effect of the drug on daily insulin doses, improvement in lipid parameters, and decline in body fat was assessed without correlating pre-specified outcomes with drug plasma concentrations.

Enhanced glucose homeostasis via Clostridium symbiosum-mediated glucagon-like peptide 1 inhibition of hepatic gluconeogenesis in mid-intestinal bypass surgery.

The small intestine is known to play a crucial role in the development and remission of diabetes mellitus (DM). However, the exact mechanism by which mid-small intestinal bypass improves glucose metabolism in diabetic rats is not fully understood. To elucidate the mechanisms by which mid-small intestinal bypass improves glucose metabolism. Streptozotocin (STZ) was used to induce DM in Sprague-Dawley (SD) rats at a dose of 60 mg/kg. The rats were then randomly divided into two groups: The mid-small intestine bypass (MSIB) group and the sham group (underwent switch laparotomy). Following a 6-wk recovery period post-surgery, the rats underwent various assessments, including metabolic parameter testing, analysis of liver glycogen levels, measurement of key gluconeogenic enzyme activity, characterization of the gut microbiota composition, evaluation of hormone levels, determination of bile acid concentrations, and assessment of the expression of the intestinal receptors Takeda G protein-coupled receptor 5 and farnesoid X receptor. The MSIB group of rats demonstrated improved glucose metabolism and lipid metabolism, along with increased hepatic glycogen content. Furthermore, there was a decrease in the expression of the key gluconeogenic enzymes phosphoenolpyruvate carboxykinase 1 and glucose-6-phosphatase. Importantly, the MSIB group exhibited a substantial increase in the abundances of intestinal Lactobacillus, Clostridium symbiosum, Ruminococcus gnavus, and Bilophila. Moreover, higher levels of secondary bile acids, such as intestinal lithocholic acid, were observed in this group. Remarkably, the changes in the gut microbiota showed a significant correlation with the expression of key gluconeogenic enzymes and glucagon-like peptide 1 (GLP-1) at 6 wk postoperatively, highlighting their potential role in glucose regulation. These findings highlight the beneficial effects of mid-small intestine bypass on glucose metabolism and the associated modulation of the gut microbiota. The findings of this study demonstrate that the introduction of postoperative intestinal Clostridium symbiosum in the mid-small intestine contributes to the enhancement of glucose metabolism in nonobese diabetic rats. This improvement is attributed to the increased inhibition of hepatic gluconeogenesis mediated by GLP-1, resulting in a favorable modulation of glucose homeostasis.

Effects of Cinnamon Powder on Glucose Metabolism in Diabetic Mice and the Molecular Mechanisms.

The liver is the primary organ regulating glucose metabolism. In our recent study, cinnamon improved liver function in diabetic mice. However, it is not clear whether cinnamon can reduce the glycemia of diabetic animals by regulating liver glucose metabolism. The purpose of this study was to investigate the hypoglycemic mechanism of cinnamon powder (CP) from the perspective of regulating liver glucose metabolism. To achieve this, different doses of CP (200, 400, or 800 mg/kg body weight) were given to diabetic mice by gavage once per day for 8 weeks. These mice were compared with healthy controls, untreated diabetic mice, and diabetic mice treated with metformin (the main first-line drug for type 2 diabetes). CP treatment effectively reduced fasting blood glucose levels and food intake, improved glucose tolerance and fasting serum insulin levels, and decreased glycated serum protein levels in diabetic mice. Furthermore, treatment with CP increased liver glycogen content and reduced the level of the gluconeogenesis precursor pyruvate in the liver. Data obtained by qPCR and western blotting suggested that CP improved glucose metabolism disorders by regulating AMPKα/PGC1α-mediated hepatic gluconeogenesis and PI3K/AKT-mediated hepatic glycogen synthesis. CP exhibits good hypoglycemic effects by improving hepatic glycogen synthesis and controlling hepatic gluconeogenesis. Therefore, CP may be applied as a functional food to decrease blood glucose.

SAT039 Chronic Glucocorticoid Exposure Induced A S1PR2-Rorc Axis To Enhance Hepatic Gluconeogenesis

Abstract Disclosure: R.A. Lee: None. M. Chang: None. A. Tsay: None. Y. Lee: None. D. Li: None. N. Yiv: None. S. Tian: None. J. Wang: None. Glucocorticoids are steroid hormones whose circulating levels are elevated in response to stress to maintain metabolic homeostasis. Glucocorticoids are also used to treat inflammatory and autoimmune diseases due to their potent anti-inflammatory and immunomodulatory activities. However, chronic glucocorticoid treatment causes various metabolic disorders, such as hyperglycemia and insulin resistance. Glucocorticoids convey their functions through an intracellular glucocorticoid receptor (GR) which is a transcriptional regulator. While GR stimulates hepatic gluconeogenic gene transcription, additional mechanisms activated specifically by chronic glucocorticoid exposure can further enhance hyperglycemia. We found that chronic glucocorticoid exposure elevated hepatic production of sphingosine-1-phosphate (S1P) and hepatic knockdown of sphingosine-1-phosphate receptor 2 (S1PR2) attenuated chronic glucocorticoid-induced glucose intolerance and hepatic gluconeogenesis while having no effect on insulin signaling. Hepatic S1PR2 knockdown reduced GR occupancy on glucocorticoid response elements (GREs) of gluconeogenic genes, such as phosphoenolpyruvate carboxykinase (Pck1) and the catalytic subunit of glucose 6 phosphatase (G6pc), thus reducing their expressions. Interestingly, RAR-related orphan receptor C (Rorc) expression was reduced in the liver of glucocorticoid treated hepatic S1PR2 knockdown mice. We found that Rorc was also recruited to the promoter of Pck1 upon glucocorticoid treatment and reducing Rorc activity decreased the GR occupancy at the GRE of Pck1. In contrast, overexpressing Rorc in the liver of hepatic S1PR2 knockdown mice restored glucocorticoid-induced glucose intolerance, GR recruitment to GREs of gluconeogenic genes, and gluconeogenic gene expression. Overall, we propose that chronic glucocorticoid exposure induces a novel S1PR2-Rorc axis to promote the transcription of gluconeogenic genes to enhance hyperglycemia. Presentation: Saturday, June 17, 2023

FRI037 Time-restricted Feeding Prevents Progression Of Nonalcoholic Fatty Liver Disease/Nonalcoholic Steatohepatitis Through IPMK

Abstract Disclosure: I. Jung: None. F. Anokye-Danso: None. S. Jin: None. R.S. Ahima: None. S. Kim: None. Nonalcoholic fatty liver disease (NAFLD) has become the most common chronic liver disease worldwide, with a broad spectrum ranging from simple steatosis to advanced stage of nonalcoholic steatohepatitis (NASH). Although many clinical trials for NAFLD treatment are undergoing, there are currently no approved therapies. Insulin resistance is central to the progression of NAFLD and implicated in disease progression from steatosis to NASH. Therefore, modulation of insulin resistance represents a potential strategy for NAFLD treatment. We have reported that inositol polyphosphate multikinase (IPMK), a key enzyme in inositol polyphosphate biosynthesis, plays a role in regulating hepatic insulin signaling and gluconeogenesis both in vitro and in vivo. However, the roles of IPMK in hepatic lipid metabolism were not investigated. To determine the role of IPMK in NAFLD/NASH, we fed high fat diet (HFD) or methionine-choline deficient diet (MCDD) in both wild type (WT) and liver-specific IPMK knockout (LKO) mice. We also examined the effect of time-restricted feeding (TRF), which is known to be beneficial for metabolic disease without influencing the food intake, in NAFDL/NASH. Here, we show that hepatic IPMK protein was decreased in WT mice fed a MCDD compared to a normal chow diet (ND) and loss of IPMK in the liver augments hepatic steatosis and steatohepatitis in mice fed a HFD as well as MCDD. Compared to the WT mice, triglycerides (TG) and pro-inflammatory gene expression in the liver increase significantly in the LKO mice fed HFD. Importantly, MCDD-fed LKO mice exhibit exacerbated NAFLD/NASH accompanied by an increase in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), hepatic TG accumulation and fibrosis, and pro-inflammatory gene expression. Finally, TRF by limiting a food access only during the dark cycle led to an increase in expression hepatic IPMK protein in MCDD mice. Importantly, TRF alleviates NAFLD/NASH reflected by reduced hepatic TG accumulation, lowered serum ALT and AST levels, downregulated pro-inflammatory gene expression compared with ad libitum mice. Our results demonstrate that loss of IPMK mediates the progression of NAFLD/NASH and may be potentially targeted for treatment of NAFLD/NASH. Presentation: Friday, June 16, 2023

OR12-03 The Pancreatic Islet Alpha Cell Arginine Transporter SLC7A2 Regulates Arginine-induced Glucagon Secretion and Glycemia

Abstract Disclosure: J.E. Stanley: None. A. Reuter: None. K. Sellick: None. A.D. Attie: None. M.P. Keller: None. E. Dean: None. Glucagon secreted from pancreatic islet α cells stimulates both hepatic glycogenolysis and gluconeogenesis resulting in the fractional extraction of amino acids from the blood. Thus, glucagon stimulates hepatic glucose output and raises blood glucose. When liver glucagon signaling is impaired, amino acid accumulation (hyperaminoacidemia) promotes subsequent α cell hyperplasia and hyperglucagonemia via this liver-α-cell axis, an endocrine feedback loop. We show that high levels of both glutamine and recently arginine are required to stimulate α cells to proliferate. Arginine also potently stimulates glucagon secretion, but the mechanisms behind this are not well defined. We find that the cationic amino acid transporter, SLC7A2/CAT2 is the most highly expressed amino acid transporter in human pancreatic α cells and three-fold higher in α cells than it is in β-cells in both mouse and human islets. We also find two SNPs in the human SLC7A2 locus are strongly associated with HbA1c (rs142010226 (A/G) - chr8: 17,367,112; p=9.11e-18; β=0.0113 and rs2517232 - chr8: 17,367,421; p=2.553e-14; β=-0.0283). Both SNPs are in the first intron of SLC7A2 but appear to exert opposite effect on HbA1c and associated with distinct haplotype blocks. ATAC-Seq analyses of human islets show that these SNPs are within 1 kb of binding sites for MAFB and FOXA2 transcription factors, both critical for α cell gene expression. To test if α cell expression of SLC7A2 plays a role in both glycemia and arginine-stimulated glucagon secretion, we generated α cell-specific Slc7a2-/- (α7A2KO) mice. Blood glucose and glucagon levels were not significantly different after a 6-hour fast in α7A2KO and 7A2WTCre+ mice. After a 6-hour fast, mice were administered an arginine bolus (2g/kg) via intraperitoneal injections and their blood glucose and glucagon levels were measured 15 minutes post-injection. α7A2KO mice had a significantly higher blood glucose level than 7A2WTcre+ mice after stimulation with arginine (arginine-stimulated blood glucose - 7A2WTCre+ 124.3 ± 26.7 mg/dl vs. α7A2KO 159.6 ± 32.1 mg/dl n=23-25 p=0.0019). However, glucagon secretion was significantly lower after stimulation with arginine in the α7A2KO mice compared to the 7A2WTCre+ mice (arginine-stimulated glucagon - 7A2WTCre+ 321.8 ± 171.9 pM vs. α7A2KO 74.6 ± 22.8 pM n=4-6; p<0.0001). Together, these data indicate that SLC7A2 mediates arginine’s effects in α cells and plays an important role in regulating glycemia. Presentation: Friday, June 16, 2023

SAT159 A Unique Case Of IGF-2 Induced Hypoglycemia As First Manifestation Of Hepatocellular Carcinoma

Abstract Disclosure: I. Ali: None. M. Alnaher: None. A. Arida: None. Introduction: Hypoglycemia is one of the manifestations of non-islet cell tumor-induced hypoglycemia (NICTH). In hepatocellular carcinoma (HCC), it has been reported in 4-27% of patients, and it is associated with poor prognosis. Case: Patient is a 70-years-old Female with past medical history of prediabetes and untreated Hepatitis C infection who presented with sever weakness and was found to have blood glucose of 37. She was started on Dextrose 10% with improvement of her symptoms. During hospital course she had recurrent episodes of symptomatic hypoglycemia. Patient denied any family history of diabetes with no access to diabetes medication. Blood work showed elevated AST, ALT with low albumin, low prealbumin and AM cortisol 13, Sulfonylureas screen is Negative. Result of 72 hours hypoglycemia protocol while blood glucose level was 48 mg/dl as follow: Proinsulin, Intact 1.6 mmol/L (Ref: less than 8.0), insulin like growth factor -1 (IGF-1): 10 ng/ml (26-226), IGF-2: 69 ng/ml (180-580) so IGF-2/IGF-1 ratio of 6.9Insulin: 0.2 uIU/ml (1.9-23), C-Peptide 0.1 ng/ml (0.5-3.3) and Beta hydroxybutyrate: 0.1 mmol/L. CT abdomen: 7.0 x 7.2 x 10.5 cm right hepatic lobe mass with extension to right portal vein that is most consistent with hepatocellular carcinoma with background liver cirrhosis with portal hypertension without ascites. Right hepatic artery tumor biopsy showed moderately differentiated HCC. Patient was started on steroid therapy then octreotide. Multidisciplinary meeting discussed in details treatment options and prognosis including treatment of her cancer with surgery vs chemoradiation therapy, but patient and her family opted for DNR/DNI, and patient passed away during that admission. Discussion: Even though HCC-related hypoglycemia is a known phenomenon, knowledge about prevention and treatment is lacking. The suggested mechanism of hypoglycemia in NICTH include increased consumption of glucose by tumor in malnourished patients with low glycogen deposits and defective gluconeogenesis or may be caused by IGF-2 overproduction that crosses capillary membranes and stimulates insulin receptor. Hypoglycemia work up reveals low levels of insulin, pro-insulin, C-peptide, and β-hydroxybutyrate. Levels of GH and IGF-1 will also be low due to the suppressive effect of IGF-2. Total IGF-2 may be elevated or normal but the ratio of IGF-2/IGF-1 is usually elevated 3:1. Management of an IGF-2 producing tumor with surgical resection, radiation, and/or chemotherapy can potentially resolve hypoglycemia. Medical therapy with glucocorticoids has been utilized by promoting hepatic gluconeogenesis and lipolysis and reducing peripheral glucose uptake. Glucocorticoids also help to decrease the levels of IGF-2 either by decreased tumor production or increased clearance. Case reports have documented the use of glucagon, growth hormone, and octreotide, however, effects are limited. Presentation: Saturday, June 17, 2023

Comprehensive quantification of metabolic flux during acute cold stress in mice

Cold-induced thermogenesis (CIT) is widely studied as a potential avenue to treat obesity, but a thorough understanding of the metabolic changes driving CIT is lacking. Here, we present a comprehensive and quantitative analysis of the metabolic response to acute cold exposure, leveraging metabolomic profiling and minimally perturbative isotope tracing studies in unanesthetized mice. During cold exposure, brown adipose tissue (BAT) primarily fueled the tricarboxylic acid (TCA) cycle with fat in fasted mice and glucose in fed mice, underscoring BAT's metabolic flexibility. BAT minimally used branched-chain amino acids or ketones, which were instead avidly consumed by muscle during cold exposure. Surprisingly, isotopic labeling analyses revealed that BAT uses glucose largely for TCA anaplerosis via pyruvate carboxylation. Finally, we find that cold-induced hepatic gluconeogenesis is critical for CIT during fasting, demonstrating a key functional role for glucose metabolism. Together, these findings provide a detailed map of the metabolic rewiring driving acute CIT.

THU382 It’s Not What It Seems - Metformin Induced Severe Lactic Acidosis Mimicking Euglycemic Diabetic Ketoacidosis

Abstract Disclosure: M. Shaukat: None. R. Memon: None. M. Mack: None. E. Galarza: None. R. Saeed: None. Diabetic Ketoacidosis (DKA) is a life-threatening complication and, occasionally, the initial presentation of Diabetes Mellitus (DM). DKA can rarely present without elevated blood glucose levels, known as Euglycemic DKA, now typically associated with SGLT-2 inhibitors. Metformin is known to cause lactic acidosis but has not been reliably associated with euglycemic DKA. We present a case of metformin-associated lactic acidosis with renal failure and ketosis mimicking euglycemic DKA. Case: 82-year-old female with a history of DM type 2 on metformin and hypertension presented to the Emergency Department (ED) with complaints of presyncope, vomiting and generalized weakness for two weeks. In the ED, her temp. was 90.3 F and BP was 66/28 mmHg. Her labs revealed a potassium of 5.4mmol/L, bicarbonate 5 mmol/L, Creatinine 3.68 mg/dL (baseline 0.7mg/dL), Anion gap (AG) 45mmol/L, Lactate 25.1 mmol/L and Beta-hydroxybutyrate of 21.4 mg/dL. CT Scan of the chest and abdomen showed a 2 cm liver mass and pancreatitis. She was taken to the ICU and started on antibiotics, IV fluids, and vasopressors. The following day, her lactate was 13.5mg/dL, AG was 37mmol/L and Beta-hydroxybutyrate increased to 53.5mg/dL. Her blood sugars remained between 140-180mg/dL and her hemoglobin A1c was in the prediabetic range at 6.1%. She was thought to be in metformin induced euglycemic diabetic ketoacidosis and was started on an insulin drip with 5% Dextrose and 0.45% Normal Saline. Nephrology was consulted and she underwent hemodialysis for metformin toxicity. She remained on the insulin drip for 48 hours after which she was successfully weaned off. She currently remains off on any antidiabetic medication and is doing well. Discussion: DKA is a life-threatening complication of DM that presents as hyperglycemia, ketonemia, and AGMA. Euglycemic DKA is distinguished by blood glucose levels less than 250mg/dL and is usually seen with pregnancy, fasting states and use of SGLT-2 inhibitors. Previous cases of metformin-associated lactic acidosis (MALA) and concurrent euglycemic DKA have been reported, out of which several patients were concurrently on SGLT-2 inhibitors. After a thorough literature review, we were unable to determine a causal mechanism linking metformin and euglycemic DKA. In the reported cases, we believe that renal failure from various causes precipitated MALA due to decrease metformin clearance, with ketosis caused by starvation and exacerbated by metformin-induced inhibition of hepatic gluconeogenesis, further aggravated by an increase in stress hormones resulting in increased insulin resistance and ultimately leading to the development of the severe ketoacidosis. Even though this phenomenon has previously been referred to as “Metformin-induced euglycemic DKA”, we believe that this is more akin to a mimic, rather than true DKA. Presentation: Thursday, June 15, 2023

SAT038 Dietary Trans-10,cis-12 Conjugated Linoleic Acid Regulates Glucose Levels In Mice With Severe Insulin Resistance: Potential Effects On The Liver Via HNF4α And FOX01

Abstract Disclosure: S.M. Lee: None. J.T. Muratalla: None. C.W. Liew: None. J. Cordoba-Chacon: None. Plasma trans-10 cis-12-conjugated linoleic acid CLA (t10c12-CLA) is inversely associated with increased risk of diabetes in humans. In mice, it is well-known that t10c12-CLA induces whole-body insulin resistance due to severe lipodystrophy (loss of whole-body adipose tissue). However, t10c12-CLA-fed mice show normal glucose levels and, t10c12-CLA improved glucose clearance in models with preexisting insulin resistance: Zucker fatty/diabetic rats, obese and insulin resistant Ldlr-/- mice, and diet-induced obese and insulin resistant male mice. In this study, we assessed whether supplementation of t10c12-CLA in a low-fat diet reduces glucose levels in adipocyte-specific insulin receptor knockout (IRFKO) mice, which is a mouse model of congenital lipodystrophy. In addition to lipodystrophy, IRFKO mice show liver steatosis and increased blood glucose levels that are consequences of whole-body insulin resistance and increased hepatic gluconeogenesis. Briefly, 5-6-week-old male and female control and IRFKO mice were fed with a low-fat diet supplemented with 0.8% t10c12-CLA for 5 weeks. In IRFKO mice, dietary t10c12-CLA did not reduce adipose tissue mass. However, dietary t10c12-CLA increased liver steatosis and plasma insulin levels. Surprisingly, dietary t10c12-CLA reduced glucose levels in both male and female IRFKO mice. Also, dietary t10c12-CLA reduced food intake and polydipsia, which were increased in IRFKO mice due to lipodystrophy and hyperglycemia, respectively. A transcriptomic analysis (RNA-seq) of livers of control and IRFKO male mice fed with a t10c12-CLA diet showed that t10c12-CLA reduced the expression of the key gluconeogenic and mitochondrial fatty oxidation genes. Previous studies have suggested that t10c12-CLA reduces the expression of Forkhead box 01 (Fox01) and Hepatocyte nuclear factor 4 alpha (HNF4α). Of note, the livers of t10c12-CLA-fed control and IRFKO male mice showed a downregulation of positive targets of FoxO1 and HNF4α. In addition, livers of t10c12-CLA-fed mice showed reduced total FOXO1 protein levels, and increased phosphorylation of FOXO1 in t10c12-CLA-fed IRFKO mice. These findings suggest that t10c12-CLA could be directly acting on the liver to alter HNF4α and FOXO1-dependent regulation of hepatic fatty acid oxidation and glucose production. These actions may explain why t10c12-CLA-fed mice do not show increased glucose levels and is potentially linked to the inverse association between t10c12-CLA levels and diabetes risk. Presentation: Saturday, June 17, 2023

THU389 Euglycemic Diabetic Ketoacidosis With Prolonged Ketosis Due To SGLT-2 Inhibitor

Abstract Disclosure: N. Viera Feliciano: None. J. Johannan: None. E. Nosova: None. C.J. Levy: Advisory Board Member; Self; Eli Lilly & Company, Dexcom. Research Investigator; Self; NIH, Helmsley Foundation, Abbott, Dexcom, Insulet, Tandem. Background: Euglycemic Diabetic ketoacidosis is a known complication of Sodium-glucose cotransporter 2 inhibitor use. Prolonged ketosis is infrequently reported on this patient population. Clinical Case: A 67-year-old male with T2DM (HbA1c 8.3%) taking dapagliflozin, metformin and semaglutide presented with emesis, polydipsia and reduced oral intake for 2 weeks. Initial tests were consistent with euglycemic diabetic ketoacidosis (eDKA): glucose 193 (70-110 mg/dL), acidosis (pH on VBG 7.27), bicarbonate 10 mmol/L (22-26 mmol/L), anion gap of 25, beta-hydroxybutyrate >4.5, positive urine ketones and glycosuria. Insulin drip was initiated along with fluid resuscitation and intravenous dextrose, followed by transition to a basal/bolus regimen after anion gap closure. Despite overall clinical improvement, the patient’s labs demonstrated persistent metabolic derangements: glycosuria and ketosis that lasted 7 days after his last dose of dapagliflozin. Implementation of an insulin regimen in this insulin-naïve patient and a carbohydrate-rich diet resulted in subsequent resolution of the derangements. The patient was discharged on metformin and insulin. eDKA is defined by a blood glucose level below 200 mg/dL, metabolic acidosis and ketosis. The cause of eDKA in this case is attributed to SGLT-2 inhibitor (SGLT2i) use, which lead to a carbohydrate deficit and depletion of glycogen stores, increased glucagon/insulin ratio resulting in ketogenesis and decreased hepatic gluconeogenesis. SGLT-2i act on proximal tubules reducing reabsorption of glucose, thereby increasing urinary glucose excretion. Dapagliflozin has an approximate half-life of 12 hours; therefore, clinical recovery from SGLT2i-associated eDKA would be expected to take up to 2-3 days after cessation of the medication. Case reports have shown persistent glucosuria and ketonemia up to 10 days after discontinuation of the drug, but the data is sparse. Our patient’s laboratory tests remained consistent with eDKA for up to 7 days, suggesting a longer lasting effect of dapagliflozin. This was managed with more aggressive insulin doses and a higher carbohydrate intake which played a critical role in correction of the insulin deficit and repletion of glycogen stores for metabolic recovery. With the growing popularity of this medication class, additional controlled studies are needed for a better understanding of the pharmacodynamics as well as how to target management strategies when eDKA occurs. In this report, we highlight the prolonged clinical effects of SGLT-2i after discontinuation and resolution of symptoms with an aggressive insulin regimen and higher carbohydrate intake. Conclusion: Use of SGLT-2i therapy for diabetes has increased the incidence of eDKA. Providers should be aware of the possibility of prolonged eDKA and manage metabolic imbalances to achieve clinical recovery in an appropriate time frame. Presentation: Thursday, June 15, 2023

Anticancer effect of metformin alone and in combination with 2-deoxy-D-glucose on mouse T cell lymphoma EL4 cells

Metformin is a treatment used widely for non-insulin-dependent diabetes mellitus with few side effects and acts by inhibiting hepatic gluconeogenesis and glucose absorption from the gastrointestinal tract. Lymphoma is one of the most common hematological malignancies in dogs. Chemotherapy is used mainly on lymphoma, but further research on developing anticancer drugs for lymphoma is needed because of its severe side effects. This study examined the anticancer effects of metformin alone and in combination with 2-deoxy-D-glucose (2-DG), a glucose analog, on EL4 cells (mouse T-cell lymphoma). Metformin reduced the metabolic activity of EL4 cells and showed an additive effect when combined with 2-DG. In addition, cell death was confirmed using a trypan blue exclusion test, Hochest 33342/propidium iodide (PI) staining, and Annexin V/PI staining. An analysis of the cell cycle and mitochondria membrane potential (MMP) to investigate the mechanism of action showed that metformin stopped the G2/M phase of EL4 cells, and metformin + 2-DG decreased MMP. Metformin exhibited anticancer effects as a G2/M phase arrest mechanism in EL4 cells and showed additive effects when combined with 2-DG via MMP reduction. Unlike cytotoxic chemotherapeutic anticancer drugs, metformin and 2-DG are related to cellular glucose metabolism and have little toxicity. Therefore, metformin and 2-DG can be an alternative to reduce the toxicity caused by chemotherapeutic anticancer drugs. Nevertheless, research is needed to verify the in vivo efficacy of metformin and 2-DG before they can be used in lymphoma treatments.

Understanding the Relationship between Nonalcoholic Fatty Liver Disease and Thyroid Disease.

The prevalence of hypothyroidism in patients with nonalcoholic fatty liver disease (NAFLD) is high (22.4%). Thyroid hormones (THs) regulate many metabolic activities in the liver by promoting the export and oxidation of lipids, as well as de novo lipogenesis. They also control hepatic insulin sensitivity and suppress hepatic gluconeogenesis. Because of its importance in lipid and carbohydrate metabolism, the involvement of thyroid dysfunction in the pathogenesis of NAFLD seems plausible. The mechanisms implicated in this relationship include high thyroid-stimulating hormone (TSH) levels, low TH levels, and chronic inflammation. The activity of the TH receptor (THR)-β in response to THs is essential in the pathogenesis of hypothyroidism-induced NAFLD. Therefore, an orally active selective liver THR-β agonist, Resmetirom (MGL-3196), was developed, and has been shown to reduce liver fat content, and as a secondary end point, to improve nonalcoholic steatohepatitis. The treatment of NAFLD with THR-β agonists seems quite promising, and other agonists are currently under development and investigation. This review aims to shine a light on the pathophysiological and epidemiological evidence regarding this relationship and the effect that treatment with THs and selective liver THR-β agonists have on hepatic lipid metabolism.

Dairy Cows Transition Period Feeding and Management

Proper feeding during the pregnancy and post-pregnancy period, determines the milk Production, ensuing lactation and as well as, the reproductive efficiency of the animal. 3 weeks before and 3 weeks after parturition are very important because a dramatic physiological and metabolic adaptation occurs during this period which is called the transition period. The Nutritional imbalances are caused by the low quality of desired nutritious rations, reduced appetite, and low ingestion which are responsible for depressed performance, specifically during the transition period. The absence of knowledge and skills of farmers in feeding management during the transition period exacerbates the situation. The metabolic changes, metabolic adaptations, Mechanism of a Transition period, transition period feeding management to prevent disorder, Decreased dry matter intake (DMI), Nutrient requirements, Infectious diseases and metabolic disorders, Nutritional management to Ease out the Transition Period, Management of the transition period, Feed additives supplementation and nutritional-management strategies to passage transition period are discussed by this review paper. Simultaneously, with the growth of the fetus, the nutrient requirements increase and are at their peak before parturition as well as immediately after parturition. The peak milk production reaches about 5–8 weeks postpartum, while the diet consumption peaks at 10–14 weeks postpartum. So, dairy cows will typically suffer a 6–8-week period of negative energy balance during the postpartum period. The requirement energy of the mammary at 4 days postpartum is more than three times than that of the uterus, with a simultaneous increase in the requirement of metabolizable protein, especially of methionine and lysine. The high metabolic incidence of infectious diseases is responsible for the high incidence of inflammatory conditions, mostly immediately after calving. To prevent mentioned diseases, pro-inflammatory cytokine release should be avoided postpartum. The body reserves mobilization, especially fat and protein; and hepatic gluconeogenesis takes place immediately after parturition leading to increased levels of β-hydroxy butyric acid and non-esterified fatty acids, which act as a gateway for several metabolic diseases. The phase of transition may eased by increasing DMI, feed conversion efficiency, density of ration, and protein supplements rich in rumen un-degradable protein; by using anionic mineral mixture, optimizing roughage to concentrate (R: C) ratio, optimum physically effective fiber length of forages/silage/hay, dietary buffers, feed additives such as inflammation modulators, protected nutrients, glucogenic precursors and direct-fed microbial/probiotics.

Mechanism of the antidiabetic action of Nigella sativa and Thymoquinone: a review.

Long used in traditional medicine, Nigella sativa (NS; Ranunculaceae) has shown significant efficacy as an adjuvant therapy for diabetes mellitus (DM) management by improving glucose tolerance, decreasing hepatic gluconeogenesis, normalizing blood sugar and lipid imbalance, and stimulating insulin secretion from pancreatic cells. In this review, the pharmacological and pharmacokinetic properties of NS as a herbal diabetes medication are examined in depth, demonstrating how it counteracts oxidative stress and the onset and progression of DM. This literature review drew on databases such as Google Scholar and PubMed and various gray literature sources using search terms like the etiology of diabetes, conventional versus herbal therapy, subclinical pharmacology, pharmacokinetics, physiology, behavior, and clinical outcomes. The efficiency and safety of NS in diabetes, notably its thymoquinone (TQ) rich volatile oil, have drawn great attention from researchers in recent years; the specific therapeutic dose has eluded determination so far. TQ has anti-diabetic, anti-inflammatory, antioxidant, and immunomodulatory properties but has not proved druggable. DM's intimate link with oxidative stress, makes NS therapy relevant since it is a potent antioxidant that energizes the cell's endogenous arsenal of antioxidant enzymes. NS attenuates insulin resistance, enhances insulin signaling, suppresses cyclooxygenase-2, upregulates insulin-like growth factor-1, and prevents endothelial dysfunction in DM. The interaction of NS with mainstream drugs, gut microbiota, and probiotics opens new possibilities for innovative therapies. Despite its strong potential to treat DM, NS and TQ must be examined in more inclusive clinical studies targeting underrepresented patient populations.

Circadian rhythm disturbances involved in ozone-induced glucose metabolism disorder in mouse liver

Ozone (O3) is a key environmental factor for developing diabetes. Nevertheless, the underlying mechanisms remain unclear. This study aimed to investigate alterations of glycometabolism in mice after O3 exposure and the role of circadian rhythms in this process. C57BL/6 male mice were randomly assigned to O3 (0.5 ppm) or filtered air for four weeks (4 h/day). Then, hepatic tissues of mice were collected at 4 h intervals within 24 h after O3 exposure to test. The results showed that hepatic circadian rhythm genes oscillated abnormally, mainly at zeitgeber time (ZT)8 and ZT20 after O3 exposure. Furthermore, detection of glycometabolism (metabolites, enzymes, and genes) revealed that O3 caused change in the daily oscillations of glycometabolism. The serum glucose content decreased at ZT4 and ZT20, while hepatic glucose enhanced at ZT16 and ZT24(0). Both G6pc and Pck1, which are associated with hepatic gluconeogenesis, significantly increased at ZT20. O3 exposure disrupted glycometabolism by increasing gluconeogenesis and decreasing glycolysis in mice liver. Finally, correlation analysis showed that the association between Bmal1 and O3-induced disruption of glycometabolism was the strongest. The findings emphasized the interaction between adverse outcomes of circadian rhythms and glycometabolism following O3 exposure.

Cold-activated brown fat-derived extracellular vesicle-miR-378a-3p stimulates hepatic gluconeogenesis in male mice

During cold exposure, activated brown adipose tissue (BAT) takes up a large amount of circulating glucose to fuel non-shivering thermogenesis and defend against hypothermia. However, little is known about the endocrine function of BAT controlling glucose homoeostasis under this thermoregulatory challenge. Here, we show that in male mice, activated BAT-derived extracellular vesicles (BDEVs) reprogram systemic glucose metabolism by promoting hepatic gluconeogenesis during cold stress. Cold exposure facilitates the selective packaging of miR-378a-3p—one of the BAT-enriched miRNAs—into EVs and delivery into the liver. BAT-derived miR-378a-3p enhances gluconeogenesis by targeting p110α. miR-378 KO mice display reduced hepatic gluconeogenesis during cold exposure, while restoration of miR-378a-3p in iBAT induces the expression of gluconeogenic genes in the liver. These findings provide a mechanistic understanding of BDEV-miRNA as stress-induced batokine to coordinate systemic glucose homoeostasis. This miR-378a-3p-mediated interorgan communication highlights a novel endocrine function of BAT in preventing hypoglycemia during cold stress.

Metformin: A New Inhibitor of the Wnt Signaling Pathway in Cancer.

The biguanide drug metformin is widely used in type 2 diabetes mellitus therapy, due to its ability to decrease serum glucose levels, mainly by reducing hepatic gluconeogenesis and glycogenolysis. A considerable number of studies have shown that metformin, besides its antidiabetic action, can improve other disease states, such as polycystic ovary disease, acute kidney injury, neurological disorders, cognitive impairment and renal damage. In addition, metformin is well known to suppress the growth and progression of different types of cancer cells both in vitro and in vivo. Accordingly, several epidemiological studies suggest that metformin is capable of lowering cancer risk and reducing the rate of cancer deaths among diabetic patients. The antitumoral effects of metformin have been proposed to be mainly mediated by the activation of the AMP-activated protein kinase (AMPK). However, a number of signaling pathways, both dependent and independent of AMPK activation, have been reported to be involved in metformin antitumoral action. Among these, the Wingless and Int signaling pathway have recently been included. Here, we will focus our attention on the main molecular mechanisms involved.

Metformin Protects Rat Skeletal Muscle from Physical Exercise-Induced Injury.

Metformin (Met) is a drug commonly prescribed in type 2 diabetes mellitus. Its efficacy is due to the suppression of hepatic gluconeogenesis, enhancement of peripheral glucose uptake and lower glucose absorption by the intestine. Recent studies have reported Met efficacy in other clinical applications, such as age-related diseases. Despite the wide clinical use of Met, its mechanism of action on muscle and its effect on muscle performance are unclear. We investigated the effects of Met combined with training on physical performance (PP) in healthy rats receiving Met for 8 weeks while undergoing daily moderate exercise. We evaluated the following: PP through graded endurance exercise test performed before the beginning of the training protocol and 48 h before the end of the training period; blood ALT, AST, LDH and CK-MB levels in order to address muscle damage; and several blood and muscle myokines and the expression of factors believed to be involved in muscle adaptation to exercise. Our data demonstrate that Met does not improve the positive effects of exercise on performance, although it protects myocytes from exercise-induced damage. Moreover, given that Met positively affects exercise-induced muscle adaptation, our data support the idea of the therapeutic application of Met when muscle function and structure are compromised.

The Warburg effect drives cachectic states in patients with pancreatobiliary adenocarcinoma.

We have studied whether the Warburg effect (uncontrolled glycolysis) in pancreatobiliary adenocarcinoma triggers cachexia in the patient. After 74 pancreatobiliary adenocarcinomas were removed by surgery, their glucose transporter-1 and four glycolytic enzymes were quantified using Western blotting. Based on the resulting data, the adenocarcinomas were equally divided into a group of low glycolysis (LG) and a group of high glycolysis (HG). Energy homeostasis was assessed in these cancer patients and in 74 non-cancer controls, using serum albumin and C-reactive protein and morphometrical analysis of abdominal skeletal muscle and fat on computed tomography scans. Some removed adenocarcinomas were transplanted in nude mice to see their impacts on host energy homeostasis. Separately, nude mice carrying tumor grafts of MiaPaCa-2 pancreatic adenocarcinoma cells were treated with the glycolytic inhibitor 3-bromopyruvate and with emodin that inhibited glycolysis by decreasing hypoxia-inducible factor-1α. Adenocarcinomas in both group LG and group HG impaired energy homeostasis in the cancer patients, compared to the non-cancer reference. The impaired energy homeostasis induced by the adenocarcinomas in group HG was more pronounced than that by the adenocarcinomas in group LG. When original adenocarcinomas were grown in nude mice, their glycolytic abilities determined the levels of hepatic gluconeogenesis, skeletal muscle proteolysis, adipose-tissue lipolysis, and weight loss in the mice. When MiaPaCa-2 cells were grown as tumors in nude mice, 3-bromopyruvate and emodin decreased tumor-induced glycolysis and cachexia, with the best effects being seen when the drugs were administered in combination. In conclusion, the Warburg effect in pancreatobiliary adenocarcinoma triggers cancer cachexia.