Helenton C. Barrena

Time sequence of the intensification of the liver glucose production induced by high-fat diet in mice

It is well established that the development of insulin resistance shows a temporal sequence in different organs and tissues. Moreover, considering that the main aspect of insulin resistance in liver is a process of glucose overproduction from gluconeogenesis, we investigated if this metabolic change also shows temporal sequence. For this purpose, a well‐established experimental model of insulin resistance induced by high‐fat diet (HFD) was used. The mice received HFD (HFD group) or standard diet (COG group) for 1, 7, 14 or 56 days. The HFD group showed increased (P < 0.05 versus COG) epididymal, retroperitoneal and inguinal fat weight from days 1 to 56. In agreement with these results, the HFD group also showed higher body weight (P < 0.05 versus COG) from days 7 to 56. Moreover, the changes induced by HFD on liver gluconeogenesis were progressive because the increment (P < 0.05 versus COG) in glucose production from l‐lactate, glycerol, l‐alanine and l‐glutamine occurred 7, 14, 56 and 56 days after the introduction of the HFD schedule, respectively. Furthermore, glycaemia and cholesterolemia increased (P < 0.05 versus COG) 14 days after starting the HFD schedule. Taken together, the results suggest that the intensification of liver gluconeogenesis induced by an HFD is not a synchronous ‘all‐or‐nothing process’ but is specific for each gluconeogenic substrate and is integrated in a temporal manner with the progressive augmentation of fasting glycaemia. Copyright

Ketogenesis evaluation in perfused liver of diabetic rats submitted to short-term insulin-induced hypoglycemia

Ketogenesis, inferred by the production of acetoacetate plus ß‐hydroxybutyrate, in isolated perfused livers from 24‐h fasted diabetic rats submitted to short‐term insulin‐induced hypoglycemia (IIH) was investigated. For this purpose, alloxan‐diabetic rats that received intraperitoneal regular insulin (IIH group) or saline (COG group) injection were compared. An additional group of diabetic rats which received oral glucose (gavage) (100 mg kg−1) 15 min after insulin administration (IIH + glucose group) was included. The studies were performed 30 min after insulin (1.0 U kg−1) or saline injection. The ketogenesis before octanoate infusion was diminished (p < 0.05) in livers from rats which received insulin (COG vs. IIH group) or insulin plus glucose (COG vs. IIH + glucose group). However, the liver ketogenic capacity during the infusion of octanoate (0.3 mM) was maintained (COG vs. IIH group and COG vs. IIH + glucose group). In addition, the blood concentration of ketone bodies was not influenced by the administration of insulin or insulin plus glucose. Taken together, the results showed that inspite the fact that insulin and glucose inhibits ketogenesis, livers from diabetic rats submitted to short‐term IIH which received insulin or insulin plus glucose showed maintained capacity to produce acetoacetate and ß‐hydroxybutyrate from octanoate. Copyright

Paradoxical increase in liver ketogenesis during long-term insulin-induced hypoglycemia in diabetic rats.

It is well established that insulin inhibits liver ketogenesis. However, during insulin-induced hypoglycemia (IIH) the release of counterregulatory hormones could overcome the insulin effect on ketogenesis. To clarify this question the ketogenic activity in livers from alloxan-diabetic rats submitted to long-term IIH was investigated. Moreover, liver glycogenolysis, gluconeogensis, ureagenesis and the production of l-lactate were measured, and its correlation with blood levels of ketone bodies (KB), l-lactate, glucose, urea and ammonia was investigated. For this purpose, overnight fasted alloxan-diabetic rats (DBT group) were compared with control non-diabetic rats (NDBT group). Long-term IIH was obtained with an intraperitoneal injection of Detemir insulin (1 U/kg), and KB, glucose, l-lactate, ammonia and urea were evaluated at 0, 2, 4, 6, 8 or 10 h after insulin injection. Because IIH was well established two hours after insulin injection this time was used for liver perfusion experiments. The administration of Detemir insulin decreased (P < 0.05) blood KB and glucose levels, but there was an increase in the blood l-lactate levels and a rebound increase in blood KB during the glucose recovery phase of IIH. In agreement with these results, the capacity to produce KB from octanoate was increased in the livers of DBT rats. Moreover, the elevated blood l-lactate levels in DBT rats could be attributed to the higher (P < 0.05) glycogenolysis when part of glucose from glycogenolysis enters glycolysis, producing l-lactate. In contrast, except glycerol, gluconeogenesis was negligible in the livers of DBT rats. Therefore, during long-term IIH the higher liver ketogenic capacity of DBT rats increased the risk of hyperketonemia. In addition, in spite of the fact that the insulin injection decreased blood KB, there was a risk of worsening lactic acidosis.

Insulin sensitivity and liver glucose production in the rat are influenced by lifetime food restriction

In both humans and rats, food restriction leads to increased insulin sensitivity and predisposition to hypoglycemia. We hypothesized that metabolic responses to hypoglycemic episodes could be altered in food-restricted rats. To test our hypothesis, plasma glucose levels and liver glucose production during insulin-induced hypoglycemia were assessed. Rats either had free access to food (FF group) or were food restricted from birth (FR group). As adults, they were subjected to insulin-induced hypoglycemia after an overnight fast. Plasma glucose was measured before (time 0) the intraperitoneal injection of insulin (1 U/kg) and at regular intervals for 300 minutes. Some FF and FR rats received oral glucose (100 mg/kg) 15 minutes after insulin injection, and the same time intervals were investigated. The FR rats showed a larger decrease and slower recovery of plasma glucose than the FF group, and this was not influenced by oral glucose. Liver glucose production from glycogenolysis and gluconeogenesis (ie, before and during the infusion of L-alanine) was higher and lower, respectively, in the FR rats than in the FF rats, either with or without oral glucose before liver perfusion. Preference for glycogenolysis could be a metabolic adaptation for the maintenance of plasma glucose levels during fasting despite lower food availability in the FR rats. However, long-term FR increased the severity of hypoglycemia and impaired plasma glucose recovery. In addition, hypoglycemia could not be prevented by glucose administration. Therefore, food restriction in individuals with intensive insulin therapy should be more rigorously examined.

Investigation of glycemia recovery with oral administration of glycerol, pyruvate, and L-lactate during long-term, insulin-induced hypoglycemia ☆

AIM The acute effect of oral administration of isolated or combined glycerol, pyruvate, and L-lactate on glycemia recovery (GR) during long-term, insulin-induced hypoglycemia (IIH) was compared. METHODS Glycemia of 24 h-fasted rats that received intraperitoneal injection (1.0 U/kg) of regular insulin (IIH group) or saline (COG group) and, 15, 150, or 165 min later, oral saline (control IIH), glycerol (100 mg/kg), pyruvate (100 mg/kg), L-lactate (100 mg/kg), or combined glycerol+pyruvate+L-lactate (each 33.3 or 100 mg/kg) was compared. In addition, for comparative purposes, a group that received glucose (100 mg/kg) was included. Glycemia was measured 180 min after insulin or saline injection. To investigate the participation of the hepatic availability of gluconeogenic substrates to GR, livers from IIH and COG rats that received physiological or supraphysiological concentrations of isolated or combined glycerol, pyruvate, and L-lactate were compared. Liver experiments were done 180 min after insulin or saline injection. RESULTS Oral glycerol, pyruvate, and L-lactate (isolated or combined) or glucose promoted GR. Moreover, the best GR was obtained with combined glycerol+pyruvate+L-lactate (100 mg/kg). In agreement, livers that received supraphysiological concentrations of glycerol, pyruvate, and L-lactate (isolated or combined) showed higher glucose release than livers that received physiological concentrations of these substances (isolated or combined). CONCLUSION The best GR obtained with combined administration of glycerol, pyruvate, and L-lactate (100 mg/kg) during long-term IIH was a consequence of the higher liver availability of these substances associated with a maintained liver ability to produce glucose from gluconeogenic substrates.

Effect of linseed oil and macadamia oil on metabolic changes induced by high‐fat diet in mice

The effects of linseed oil (LO) and macadamia oil (MO) on the metabolic changes induced by a high‐fat diet (HFD) rich in saturated fatty acid were investigated. For the purpose of this study, the vegetable oil present in the HFD, i.e. soybean oil (SO) was replaced with LO (HFD‐LO) or MO (HFD‐MO). For comparative purposes, a group was included, which received a normal fat diet (NFD). Male Swiss mice (6‐week old) were used. After 14 days under the dietary conditions, the mice were fasted for 18 h, and experiments were then performed. The HFD‐SO, HFD‐LO and HFD‐MO groups showed higher glycaemia (p < 0.05 versus NFD). However, no significant effect was observed on glycaemia, liver gluconeogenesis and liver ketogenesis when SO was replaced by either LO or MO. The body weight and the sum of epididymal, mesenteric, retroperitoneal and inguinal fat weights were higher (p < 0.05) in the HFD‐SO and HFD‐MO groups as compared with the NFD group. However, there was no significant difference in these parameters between the NFD and HFD‐LO groups. Thus, the protective role of LO on lipid accumulation induced by an HFD rich in saturated fatty acid is potentially mediated by the high content of ɷ‐3 polyunsaturated fatty acid in LO. Copyright