Activities Of Gluconeogenic Enzymes Research Articles

Hyperpolarized [2-13C, 3-2H3]Pyruvate Detects Hepatic Gluconeogenesis In Vivo.

The feasibility of hyperpolarized [2-13C, 3-2H3]pyruvate for probing gluconeogenesis in vivo was investigated in this study. Whereas hyperpolarized [1-13C]pyruvate has clear access to metabolic pathways that convert pyruvate to lactate, alanine, and bicarbonate, its utility for assessing pyruvate carboxylation and gluconeogenesis has been limited by technical challenges, including spectral overlap and an obscure enzymatic step that decarboxylates the labeled carbon. To achieve unambiguous detection of gluconeogenic products, the carbonyl carbon in pyruvate was labeled with 13C. To prolong the T1 relaxation time, [2-13C, 3-2H3]pyruvate was synthesized and dissolved with D2O after dynamic nuclear polarization. The T1 of [2-13C, 3-2H3]pyruvate in D2O could be improved by 76.9% (79.6 s at 1 T and 74.5 s at 3 T) as compared to [2-13C]pyruvate in water. Hyperpolarized [2-13C, 3-2H3]pyruvate with D2O dissolution was applied to rat livers in vivo under normal feeding and fasting conditions. A gluconeogenic product, [2-13C]phosphoenolpyruvate, was observed at 149.9 ppm from fasted rats only, highlighting the utility of [2-13C, 3-2H3]pyruvate in detecting key gluconeogenic enzyme activities such as pyruvate carboxylase and phosphoenolpyruvate carboxykinase in vivo.

Regulation of carbohydrate metabolism and insulin secretion in diabetic rats following treatment with Hypericum perforatum L. hairy root aqueous methanol extract

Hairy root (HR) cultures of Hypericum perforatum L. have shown promise in controlling hyperglycemia, regulating blood lipid and enzyme profiles, and improving metabolic function in vivo. These benefits are mainly attributed to the phenolic acids, flavonoids, and xanthones present in HR extracts. However, the specific mechanism underlying these effects remains unclear. This study was performed to elucidate the biochemical and molecular mechanisms driving HR the antihyperglycemic and antidiabetic effects of HR extracts. HR extract (200 mg/kg body weight) was administered daily for 14 days to healthy rats and rats with streptozotocin-induced diabetes, with glibenclamide serving as a positive control. The phenolic composition of the HR extracts was confirmed through high-performance liquid chromatography/diode array detection/electrospray ionization mass spectrometry (HPLC/DAD/ESI-MSn ) analysis. The results showed that HR extract treatment increased the plasma insulin level and pancreatic poly (adenosine diphosphate-ribose) polymerase (PARP) activity in diabetic rats, thus normalizing blood glucose levels. Additionally, it reduced the activity of gluconeogenic enzymes, increased the activity of glycolytic enzymes, and normalized the glycogen content in the liver. HR extract-treated rats also exhibited increased hepatic adenosine monophosphate-activated protein kinase (AMPK) mRNA expression and a decreased protein kinase Cε (PKCε) concentration. In conclusion, HR extract demonstrated insulinotropic effects and effectively regulated hepatic carbohydrate metabolism in diabetic rats by modulating AMPK expression and the PKCε concentration. These findings suggest the potential use of HR extract as an herbal medicine for diabetes treatment and a source of antidiabetic drug development.

Leaf extracts of Euclea natalensis A.D.C ameliorate biochemical abnormalities in high-fat-low streptozotocin-induced diabetic rats through modulation of the AMPK-GLUT4 pathway

ABSTRACT Euclea natalensis is a commonly used plant in traditional African medicine for treating several diseases, including diabetes. The objective of the present study was to evaluate the antidiabetic effects of the leaf extracts of Euclea natalensis and elucidate some of its mechanisms of action. High-fat diet – low Streptozotocin-induced diabetic Sprague Dawley rats were orally treated with the methanol and dichloromethane extracts (100 and 200 mg/kg bw), and their fasting blood glucose levels, body weights, and food and water intake were measured every seven days for 21 days. Biomarkers of oxidative stress, lipid profile, gluconeogenic enzyme activities, glucose transporter 4 (GLUT 4) protein expression, and AMP-activated protein kinase (AMPK) gene expression were also determined. The 200 mg/kg bw of methanol extracts significantly reduced fasting plasma glucose, glycated hemoglobin (HbAc1), triglycerides, and low-density lipoprotein levels. The extract increased insulin sensitivity and hepatic glycogen synthesis, decreased hepatic glucose output, and increased the activities of antioxidant enzymes in diabetic animals after 21 days of treatment. The methanol extract also significantly upregulated the relative expression of GLUT 4 protein via the promoted expression of skeletal muscle and hepatic AMPK. Histological investigation showed ameliorated pancreatic degeneration and expedited regeneration of islets of Langerhans in diabetic rats treated with the methanol extract compared with the dichloromethane extract. The findings demonstrated that Euclea natalensis has beneficial effects on the biochemical and cell abnormalities of type 2 diabetes, partly via the modulation of the AMPK – GLUT 4 pathway.

LONP1 ameliorates liver injury and improves gluconeogenesis dysfunction in acute-on-chronic liver failure.

Acute-on-chronic liver failure (ACLF) is a severe liver disease with complex pathogenesis. Clinical hypoglycemia is common in patients with ACLF and often predicts a worse prognosis. Accumulating evidence suggests that glucose metabolic disturbance, especially gluconeogenesis dysfunction, plays a critical role in the disease progression of ACLF. Lon protease-1 (LONP1) is a novel mediator of energy and glucose metabolism. However, whether gluconeogenesis is a potential mechanism through which LONP1 modulates ACLF remains unknown. In this study, we collected liver tissues from ACLF patients, established an ACLF mouse model with carbon tetrachloride (CCl 4 ), lipopolysaccharide (LPS), and D-galactose (D-gal), and constructed an in vitro hypoxia and hyperammonemia-triggered hepatocyte injury model. LONP1 overexpression and knockdown adenovirus were used to assess the protective effect of LONP1 on liver injury and gluconeogenesis regulation. Liver histopathology, biochemical index, mitochondrial morphology, cell viability and apoptosis, and the expression and activity of key gluconeogenic enzymes were detected to explore the underlying protective mechanisms of LONP1 in ACLF. We found that LONP1 and the expressions of gluconeogenic enzymes were downregulated in clinical ACLF liver tissues. Furthermore, LONP1 overexpression remarkably attenuated liver injury, which was characterized by improved liver histopathological lesions and decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in ACLF mice. Moreover, mitochondrial morphology was improved upon overexpression of LONP1. Meanwhile, the expression and activity of the key gluconeogenic enzymes were restored by LONP1 overexpression. Similarly, the hepatoprotective effect was also observed in the hepatocyte injury model, as evidenced by improved cell viability, reduced cell apoptosis, and improved gluconeogenesis level and activity, while LONP1 knockdown worsened liver injury and gluconeogenesis disorders. We demonstrated that gluconeogenesis dysfunction exists in ACLF, and LONP1 could ameliorate liver injury and improve gluconeogenic dysfunction, which would provide a promising therapeutic target for patients with ACLF.

Effect of Tetrahydrocurcumin Compared to Curcumin in Carbohydrate Metabolism and Glycoprotein Components in Type 2 Diabetes - Systematic Review

Pancreatic β-cell dysfunction and insulin resistance are the two hallmarks of type 2 diabetes mellitus. Treatment of diabetes without any side effects is still a challenge to the medical system. There is an increasing demand by patients to use the natural products with antidiabetic activity, because insulin and oral hypoglycemic drugs are having so many side effects. Curcumin is a biologically active component isolated from the rhizome of Curcuma longa that possess antidiabetic and has been proven scientifically to possess high antioxidant activity and anticancer properties. Tetrahydrocurcumin (THC) is a major curcuminoid metabolite of curcumin, naturally occurring in turmeric. The interest in THC research is increasing because it is superior to curcumin in its solubility in water, chemical stability, bioavailability, and anti-oxidative activity. Many in vitro and in vivo studies have revealed that THC exerts antidiabetic effects through various mechanisms, including modulation of oxidative stress, xenobiotic detoxification, inflammation, proliferation, metastasis, programmed cell death, and immunity. The activities of glycolytic enzymes such as hexokinase and glucose-6-phosphate dehydrogenase were found to be decreased whereas the activities of gluconeogenic enzymes (glucose-6-phosphatase and fructose-1,6-bisphosphatase) and polyol pathway enzyme-sorbitol dehydrogenase were significantly increased in diabetic control rats. In addition, the oligosaccharide moieties of glycoproteins (hexose, hexosamine, fucose and sialic acid) were also significantly increased in plasma and tissues of diabetic control rats. THC and curcumin administration to diabetic rats significantly reversed the above changes when compared to diabetic control rats. In diabetic controls, hepatic and skeletal muscle glycogen content was decreased significantly as compared to non-diabetic controls. Treatment with THC and curcumin increased the hepatic and skeletal muscle glycogen significantly. The antidiabetic and antioxidant effects of THC are more potent than those of curcumin at the same dose. The antihyperglycemic action of THC might be mediated via an enhancement of insulin action, as it is evidenced by the increased levels of insulin in diabetic rats treated with THC, which may be responsible for the reversal of changes in carbohydrate enzymes and glycoprotein components. The THC administration showed more effective than curcumin.

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.

Inhibition of Gluconeogenesis by Boldine in the Perfused Liver: Therapeutical Implication for Glycemic Control.

The alkaloid boldine occurs in the Chilean boldo tree (Peumus boldus). It acts as a free radical scavenger and controls glycemia in diabetic rats. Various mechanisms have been proposed for this effect, including inhibited glucose absorption, stimulated insulin secretion, and increased expression of genes involved in glycemic control. Direct effects on glucose synthesis and degradation were not yet measured. To fill this gap, the present study is aimed at ensuring several metabolic pathways linked to glucose metabolism (e.g., gluconeogenesis) in the isolated perfused rat liver. In order to address mechanistic issues, energy transduction in isolated mitochondria and activities of gluconeogenic key enzymes in tissue preparations were also measured. Boldine diminished mitochondrial ROS generation, with no effect on energy transduction in isolated mitochondria. It inhibited, however, at least three enzymes of the gluconeogenic pathway, namely, phosphoenolpyruvate carboxykinase, fructose-bisphosphatase-1, and glucose 6-phosphatase, starting at concentrations below 50 μM. Consistently, in the perfused liver, boldine decreased lactate-, alanine-, and fructose-driven gluconeogenesis with IC50 values of 71.9, 85.2, and 83.6 μM, respectively. Conversely, the compound also increased glycolysis from glycogen-derived glucosyl units. The hepatic ATP content was not affected by boldine. It is proposed that the direct inhibition of hepatic gluconeogenesis by boldine, combined with the increase of glycolysis, could be an important event behind the diminished hyperglycemia observed in boldine-treated diabetic rats.

Alkaloid extract of Lannea egregia leaf inhibits carbohydrate digestive enzymes and modulates gluconeogenic and nucleotide-hydrolyzing enzyme activities in oxidative isolated psoas muscles injury

Alkaloid extract of Lannea egregia leaf inhibits carbohydrate digestive enzymes and modulates gluconeogenic and nucleotide-hydrolyzing enzyme activities in oxidative isolated psoas muscles injury

Resveratrol biotransformation and actions on the liver metabolism of healthy and arthritic rats

Aimsto investigate the effects of resveratrol on glycogen catabolism and gluconeogenesis in perfused livers of healthy and arthritic rats. The actions of resveratrol-3-O-glucuronide (R3G) and the biotransformation of resveratrol into R3G was further evaluated in the livers. Main methodsarthritis was induced with Freund's adjuvant. Resveratrol at concentrations of 10, 25, 50, 100 and 200 μM and 200 μM R3G were introduced in perfused livers. Resveratrol and metabolites were measured in the outflowing perfusate. Respiration of isolated mitochondria and activity of gluconeogenic enzymes were also evaluated in the livers. Key findingsresveratrol inhibited glycogen catabolism when infused at concentrations above 50 μM and gluconeogenesis even at 10 μM in both healthy and arthritic rat livers, but more sensitive in these latter. Resveratrol above 100 μM inhibited ADP-stimulated respiration and the activities of NADH- and succinate-oxidases in mitochondria, which were partially responsible for gluconeogenesis inhibition. Pyruvate carboxylase activity was inhibited by 25 μM resveratrol and should inhibit gluconeogenesis already at low concentrations. Resveratrol was significantly metabolized to R3G in healthy rat livers, however, R3G formation was lower in arthritic rat livers. The latter must be in part a consequence of a lower glucose disposal for glucuronidation. When compared to resveratrol, R3G inhibited gluconeogenesis in a lower extension and glycogen catabolism in a higher extension. Significancethe effects of resveratrol and R3G tended to be transitory and existed only when the resveratrol is present in the organ, however, they should be considered because significant serum concentrations of both are found after oral ingestion of resveratrol.

Irisin improves adiposity and exercise tolerance in a rat model of postmenopausal obesity through enhancing adipo-myocyte thermogenesis

The prevalence of obesity and its associated metabolic disorders, along with their healthcare costs, is rising exponentially. Irisin, an adipomyokine, may serve as a critical cross-organ messenger, linking skeletal muscle with adipose tissue and the liver to integrate the energy homeostasis under diet-induced obesity. We aimed to explore the putative role of irisin in the protection against obesity in a postmenopausal rat model by modulating energy expenditure (EE). Bilateral ovariectomy (OVX) was performed. After 3 weeks of recovery, the OVX rats were classified according to their dietary protocol into rats maintained on normal diets (ND) (OVX) or high-fat diet (HFD) groups. The HFD-fed animals were equally divided into OVX/HFD, or irisin-treated OVX/HFD groups. Sham rats, maintained on ND, were selected as the control group. We evaluated anthropometric, EE, and molecular biomarkers of browning and thermogenesis in inguinal white adipose tissue and skeletal muscle, and the activity of the proteins related to mitochondrial long chain fatty acid transport, oxidation, and glycolysis. HFD of OVX further deteriorated the disturbed glucose homeostasis, lipid profile, and the reduced irisin, thermogenic parameters in adipose tissue and skeletal muscle, and EE. Irisin treatment improved the lipid profile and insulin resistance. That was associated with reduced hepatic gluconeogenic enzyme activities and restored hepatic glycogen content. Irisin reduced ectopic lipid infiltration. Irisin augmented EE by activating non-shivering thermogenesis in muscle and adipose tissues and decreasing metabolic efficiency. Our experimental evidence suggests irisin’s use as a potential thermogenic agent, therapeutically targeting obesity in postmenopausal patients.Graphical abstractIrisin modulates the non-shivering thermogenesis in skeletal muscle and adipose tissue in postmenopausal model

The Role of Palmitoleic Acid in Regulating Hepatic Gluconeogenesis through SIRT3 in Obese Mice

Hepatic gluconeogenesis is a crucial process to maintain glucose level during starvation. However, unabated glucose production in diabetic patients is a major contributor to hyperglycemia. Palmitoleic acid is a monounsaturated fatty acid (16:1n7) that is available from dietary sources. Palmitoleic acid exhibits health beneficial effects on diabetes, insulin resistance, inflammation, and metabolic syndrome. However, the mechanism by which palmitoleate reduces blood glucose is still unclear. SIRT3 is a key metabolism-regulating NAD+-dependent protein deacetylase. It is known that fasting elevates the expression of SIRT3 in the liver and it regulates many aspects of liver’s response to nutrient deprivation, such as fatty acid oxidation and ketone body formation. However, it is unknown whether SIRT3 also regulates gluconeogenesis. Our study revealed that palmitoleic acid reduced hepatic gluconeogenesis and the expression of SIRT3 under high-fat diet conditions. Overexpression of SIRT3 in the liver and hepatocytes enhanced gluconeogenesis. Further study revealed that SIRT3 played a role in enhancing the activities of gluconeogenic enzymes, such as PEPCK, PC, and MDH2. Therefore, our study indicated that under a high-fat diet, palmitoleic acid decreased gluconeogenesis by reducing enzymatic activities of PEPCK, PC, and MDH2 by down-regulating the expression of SIRT3.

Biometric Indices, Physio-Metabolic Responses and Carcass Quality in Rohu (Labeo rohita) during Feed Deprivation.

Simple SummaryFeed deprivation is common among animals, including the aquatic animals. Because of this phenomenon, organisms may even suffer from severe metabolic disorders. In the present study, a rapid depletion in carbohydrate and lipid stores was noted in rohu fish (Labeo rohita), together with an elevation in gluconeogenic and antioxidant enzyme activities and inhibition of lipogenic enzyme activity under a diet-deprived condition. The biometric indices, physio-metabolic responses, and carcass quality in the rohu suggest that feed deprivation beyond 30 days caused an overall reduction in their metabolic activities. This is an interesting study, especially for aquatic habitats and aquatic life which may suffer from natural disasters, climate change impacts, or human activities altering their physio-metabolic functions.Understanding changes in biometric indices and metabolism in fish exposed to feed deprivation may be useful in aquaculture. The present study elucidates the effect of feed deprivation on physio-biochemical responses, such as changes in biometric indices, nutrient mobilization patterns, and enzyme activities in rohu (Labeo rohita). Experimental fish (av. wt. 3.41 ± 0.07 g) were deprived of feed and sampled at intervals of 0, 15, 30, 45, and 60 days to measure weight, length, body composition, and the activities of enzymes involved in digestion, metabolism, and antioxidation. A decrease in body weight, condition factor (CF), hepatosomatic index (HSI), and gastrosomatic index (GSI) was observed during the initial stage of feed deprivation (15 to 30 days) but remained unchanged thereafter. The total carbohydrate and lipid content also decreased rapidly up until 30 days, then stabilized. However, the reduction in tissue protein content (% wet weight) continued gradually with the duration of feed deprivation from 12.85 ± 0.36 at 0 days to 10.04 ± 0.67 at 15 days, 8.79 ± 0.59 at 30 days, 6.95 ± 0.69 at 45 days, and 6.16 ± 0.8 at 60 days, which was lower, compared to the other two body constituents. Amylase, protease. and lipase activities significantly reduced up until 30 days, but then stabilized. Although G6PDH enzyme activity decreased, gluconeogenic (LDH, AST, and ALT) and antioxidative (SOD and catalase) enzyme activities increased during initial feed deprivation (up to 30 days). A tissue-specific difference in amino acid metabolism with a major role of ALT in liver and AST in muscle was observed. This study revealed that rohu fingerlings adapted well to feed deprivation up until 30 days, beyond which there was an overall deterioration in the metabolic functions.

Alanine alters the carbohydrate metabolism of rainbow trout: glucose flux and cell signaling.

In rainbow trout, dietary carbohydrates are poorly metabolized compared with other macronutrients. One prevalent hypothesis suggests that high dietary amino acid levels could contribute to the poor utilization of carbohydrates in trout. In mammals, alanine is considered an important gluconeogenic precursor, but has recently been found to stimulate AMP-activated protein kinase (AMPK) to reduce glucose levels. In trout, the effect of alanine on glucose flux is unknown. The goal of this study was to determine the effects of 4 h exogenous alanine infusion on glucose metabolism in rainbow trout. Glucose flux, and the rate of glucose appearance (Ra) and disposal (Rd) were measured in vivo. Key glycolytic and gluconeogenic enzyme expression and activity, and cell signaling molecules relevant to glucose metabolism were assessed in the liver and muscle. The results show that alanine inhibits glucose Ra (from 13.2±2.5 to 7.3±1.6 μmolkg-1 min-1) and Rd (from 13.2±2.5 to 7.4±1.5 μmolkg-1 min-1) and the slight mismatch between Ra and Rd caused a reduction in glycemia, similar to the effects of insulin in trout. The reduction in glucose Rd can be partially explained by a reduction in glut4b expression in red muscle. In contrast to mammals, trout alanine-dependent glucose-lowering effects did not involve hepatic AMPK activation, suggesting a different mechanistic basis. Interestingly, protein kinase B (AKT) activation increased only in muscle, similar to effects observed in insulin-infused trout. We speculate that alanine-dependent effects were probably mediated through stimulation of insulin secretion, which could indirectly promote alanine oxidation to provide the needed energy.

Contribution of fasting and postprandial glucose-lowering mechanisms to the acute hypoglycemic effect of traditionally used Eryngium cymosum F.Delaroche

Ethnopharmacological relevanceEryngium cymosum F. Delaroche was detected as a traditional remedy against type 2 diabetes consumed by patients of Tlanchinol in the state of Hidalgo, Mexico. Aim of the study. Assessing the hypoglycemic effect and safety of the traditional extract of E. cymosum and relating it to key glucose-lowering mechanisms both in fasting and postprandial state. Materials and methodsThe aqueous extract of E. cymosum was subjected to HPLC analysis to identify its main components. Hyperglycaemic STZ-NA Wistar rats were administered with the extract to evaluate its effect on blood glucose levels and a possible dose-dependence. Afterward, it was evaluated in both pyruvate and maltose tolerance tests in STZ-NA rats to characterize its effect on gluconeogenesis and carbohydrate breakdown, two of the main mechanisms responsible for fasting and postprandial hyperglycaemia in type 2 diabetes patients. In addition, the inhibitory capacity of the extract was evaluated on key enzymes involved in gluconeogenesis and a-glucosidases. Moreover, insulin concentrations were measured in normoglycemic rats in both conditions to establish a link between the hypoglycaemic effect of the extract with insulin release and functioning. ResultsCaffeic acid (1), chlorogenic acid (2), and rosmarinic acid (3) were identified as the main constituents of the aqueous extract of E. cymosum, which exerted a hypoglycaemic effect in hyperglycaemic STZ-NA rats. It has a significant antihyperglycemic effect in the pyruvate tolerance test, and it was able to reduce the postprandial hyperglycaemia in maltose tolerance tests significantly. Moreover, it effectively reduced the activity of both gluconeogenic enzymes reaching almost 100% of inhibition, while it presented a modest 32% inhibition of aglucosidases. On the other hand, the extract decreased insulin levels after its oral administration in healthy rats in both nutritional states, without affecting normoglycemia in normal curves and reducing the postprandial peak in glucose load curves. ConclusionsThe traditional consumed form of aerial parts of E. cymosum is safe and regulated glucose levels both in fasting and in postprandial state.

Leaf extracts of Glyphaea brevis attenuate high blood glucose and lipids in diabetic rats induced with streptozotocin

Background: Diabetes is a non-communicable disease causing impairment of carbohydrate and lipid metabolisms. Previous ethnobotanical surveys showed that leaves of Glyphaea brevis (G. brevis) are used conventionally in the treatment of diabetes with limited scientific evidence. Objective: We aimed to determine the effects of aqueous (AE) and hydroethanolic extracts (HEE) of G. brevis leaves in attenuating diabetes-linked hyperglycemia and hyperlipidemia in an animal model. Materials and Methods: Thirty streptozotocin-induced diabetic male rats were divided into six groups receiving the following daily treatments for 4 weeks orally: control (distilled water), reference (tolbutamide 80 mg/kg) and 4 tests (AE 250 mg/kg, AE 500 mg/kg, HEE 250 mg/kg, and HEE 500 mg/kg). The effects of each treatment on postprandial hyperglycemia were assessed using oral glucose tolerance and oral starch tolerance tests. Blood was collected to assess the effects of treatments on fasting plasma glucose, glycated hemoglobin (HbA1c), and lipid profile. Liver glycogen and gluconeogenic enzyme activity were also measured. Results: AE- and HEE-treated rats had 36%–64% lower fasting blood glucose levels, 34%–73% lower postprandial glycemia, and 15%–75% lower HbA1c than rats from control group (P

Costus afer modulates the activities of glycolytic and gluconeogenic enzymes in streptozotocin induced diabetic rats

Background: Glycolytic enzymes (Hexokinase and α-glucosidase) and gluconeogenic enzymes (Glucose-6-phosphatase and fructose-1,6-disphosphatase) control carbohydrate metabolism and hence glucose hemostasis, an important approach for the control of diabetes. The present study was carried out to evaluate the effect of hydro-ethanolic leaf extract of Costus afer on these enzymes.
 Materials and methods: Streptozotocin-induced diabetic rats were administered hydro-ethanolic leaf extract of Costus afer (250, 500 and 1000 mg/kg body weight) orally for 8 weeks. Effect of the extract was monitored on the decrease in glycemia and the modulatory activities on glycolytic and gluconeogenic enzymes activities. The pancreas was also immunohistochemically examined for regeneration of the Islet cells.
 Results: Hydro-ethanolic extract significantly decreased the blood glucose concentration, serum glucose-6-phosphatase and fructose-1,6-disphosphatase activities which were associated with an increase in the activity of hexokinase in the liver and kidney. immunohistochemical examination of the pancreas, revealed the preservation and regeneration of the islet of the Langerhans cells. 
 Conclusion: Thus, C. afer acts against hyperglycemia modulating the activities of glucose-6-phosphatase, fructose-1,6-bisphosphatase and hexokinase, and regeneration of β-cells.

Umbelliferone stimulates glucose uptake; modulates gluconeogenic and nucleotide-hydrolyzing enzymes activities, and dysregulated lipid metabolic pathways in isolated psoas muscle

• Umbelliferone stimulated muscle glucose uptake. • Umbelliferone stalled oxidative stress in psoas muscles. • Umbelliferone modulated the enzymic activities in psoas muscles. • Umbelliferone modulated glucose uptake in psoas muscles. The ability of umbelliferone to stimulate muscle glucose uptake and its effect on glucose and fatty acid metabolism as well as nucleotide hydrolysis and cholinergic activity were investigated in isolated psoas muscles. Incubation of umbelliferone with muscles led to increase glucose uptake, elevated GSH, SOD and catalase activities, as well as depleted malondialdehyde and nitric oxide levels. It also led to decrease ATPase activity with concomitant increase in ENTPDase and 5′nucleotidiase activities. The activities of chymotrypsin and acetylcholinesterase, glycogen phosphorylase, glucose-6-phosphatase, fructose-1,6-phosphatase and lipase were also suppressed. Umbelliferone reversed the altered lipid metabolites, and deactivated pathways induced by incubation with glucose. These were however reversed in umbelliferone-incubated muscles with concomitant deactivation of steroid biosynthesis. Taken together, these results further support the antidiabetic properties of umbelliferone. Its antidiabetic mechanism involves stimulation of muscle glucose uptake, with concomitant modulation of lipid metabolism and nucleotide hydrolysis, while stalling gluconeogenesis and oxidative stress.

Phosphoenolpyruvate carboxykinase in urine exosomes reflect impairment in renal gluconeogenesis in early insulin resistance and diabetes.

Impaired insulin-induced suppression of renal gluconeogenesis could be a risk for hyperglycemia. Diabetes is associated with elevated renal gluconeogenesis; however, its regulation in early insulin resistance is unclear in humans. A noninvasive marker of renal gluconeogenesis would be helpful. Here, we show that human urine exosomes (uE) contain three gluconeogenic enzymes: phosphoenolpyruvate carboxykinase (PEPCK), fructose 1,6-bisphosphatase, and glucose 6-phosphatase. Their protein levels were positively associated with whole body insulin sensitivity. PEPCK protein in uE exhibited a meal-induced suppression. However, subjects with lower insulin sensitivity had blunted meal-induced suppression. Also, uE from subjects with prediabetes and diabetic rats had higher PEPCK relative to nondiabetic controls. Moreover, uE-PEPCK was higher in drug-naïve subjects with diabetes relative to drug-treated subjects with diabetes. To determine whether increased renal gluconeogenesis is associated with hyperglycemia or PEPCK expression in uE, acidosis was induced in rats by 0.28 M NH4Cl with 0.5% sucrose in drinking water. Control rats were maintained on 0.5% sucrose. At the seventh day posttreatment, gluconeogenic enzyme activity in the kidneys, but not in the liver, was higher in acidotic rats. These rats had elevated PEPCK in their uE and a significant rise in blood glucose relative to controls. The induction of gluconeogenesis in human proximal tubule cells increased PEPCK expression in both human proximal tubules and human proximal tubule-secreted exosomes in the media. Overall, gluconeogenic enzymes are detectable in human uE. Elevated PEPCK and its blunted meal-induced suppression in human urine exosomes are associated with diabetes and early insulin resistance.

Bacoside-A diminishes liver functional enzymes and improves carbohydrate metabolic key enzymes in streptozotocin a rat model of T2DM

Abstract Type 2 diabetes mellitus (T2DM) is a global alarming predominant metabolic disease than other diabetes and incurs substantial burden to public and healthcare systems. Our study is designed to endeavor the valuable effect of bacoside-A on hepatic key enzymes of carbohydrate metabolism in streptozotocin (STZ) induced diabetic rats. A single intraperitoneal (i.p) injection of STZ (40 mg/kg body weight [BW]) is administered to adult male Wistar rats for the attainment of diabetes. Diabetic rats were orally treated with bacoside-A of various doses (5, 10 and 20 mg/kg BW) for 45 days. Bacoside-A (10 mg/kg BW) showed more pronounced effect than the other doses and thus resulted in a significant reduction in the level of plasma glucose, glycosylated hemoglobin (HbA1c) and an rise in insulin and hemoglobin levels. Administration of bacoside-A showed significant elevated in the activity of glycolytic enzyme (hexokinase) and hepatic shunt enzyme (glucose-6-phophate dehydrogenase) whereas significant decline in the activity of gluconeogenic enzymes (glucose-6-phosphatase and fructose-1,6-bisphosphatase) in diabetic rats. Further, bacoside-A administration to diabetic rats enhanced body weight, liver glycogen content and modulating its metabolizing enzymes (glycogen synthase and glycogen phosphorylase) demonstrated the antihyperglycemic potential of bacoside-A in diabetic rats. Furthermore, hepatic marker enzymes such as aspartate transaminases (AST) and alanine transaminases (ALT) activities significantly decreased in diabetic rats and tend to normalcy in bacoside-A treated diabetic rats. Glibenclamide, a reference drug. In conclusion, our data explicated the bacoside-A, a promising antidiabetic agent which infers in modulating hepatic key enzymes by preventing defect in carbohydrate metabolism against T2DM.

Production of bioactive compounds with bactericidal and antioxidant potential by endophytic fungus Alternaria alternata AE1 isolated from Azadirachta indica A. Juss.

For combating multidrug-resistant microorganisms, exploration of natural compounds from plant endophytes increases the chance of finding novel compounds. An efficient bioactive metabolites producing endophytic fungal strain AE1 was isolated from leaves of Azadirachta indica A. Juss. The metabolites were found to be thermostable, non-proteinacious and produced prominent zones of inhibition against numbers of Gram positive and Gram negative bacteria. Based on 28S rDNA (D1/D2) sequence homology the isolate AE1 was identified as Alternaria alternata. Malt extract broth was found effective for the maximum production of bioactive metabolites by the isolate and was subjected for solvent extraction. The Ethyl acetate (EA) fraction of AE1 showed MIC values of 300–400 μg/ml against Gram positive and Gram negative bacteria tested. The cidal mode of action of EA fraction was detected by treating bacterial cultures at mid log phase. Scanning electron microscopic study supported morphological disintegration of bacterial cells. Release of nucleic acid, protein and potassium ions (K+) also suggested lysis of bacterial cells or leakage of cell membrane upon treatment. In addition, reduction of the activity of EMP pathway, TCA cycle and gluconeogenic enzymes in all bacteria suggested the interference of antibacterial principles with central carbohydrate metabolic pathways. Thin layer chromatographic separation followed by GC-MS analysis of EA fraction suggested numbers of antimicrobial compound production by AE1. In addition, DPPH free radical as well as superoxide radical scavenging assay also suggested strong antioxidant potential of AE1 with an IC50 value of 38.0±1.7 μg/ml and 11.38±1.2 μg/ml respectively. On the basis of above facts it can be concluded that the strain AE1 will be a good source of bioactive compounds having medicinal importance.