Meal-induced insulin sensitization in conscious and anaesthetized rat models comparing liquid mixed meal with glucose and sucrose.

We have recently shown that meal-induced insulin sensitization (MIS) occurs after feeding and decreases progressively to insignificance after 24 h of fasting and is caused by action of a hepatic insulin sensitizing substance (HISS). In order to carry out quantitative studies of MIS, some standardized meal intake is required. Our objective was to establish animal models to be tested in both the conscious and anaesthetized state using intragastric injection of liquid meals in order to quantify MIS. Insulin sensitivity was assessed before and 90 min after the meal using the rapid insulin sensitivity test (RIST) which is a transient euglycaemic clamp. Rats tested in the conscious state were instrumented under anaesthesia 6-9 d prior to testing with catheters in the carotid artery, jugular vein and stomach. Meals, injected into the stomach, consisted of a liquid mixed meal, sucrose, glucose or water. The glucose sequestration in response to insulin increased by 90 % and 61 % following the liquid mixed meal (10 ml/kg) in conscious and anaesthetized rats, respectively. Glucose, sucrose and water did not effectively activate MIS. MIS was completely reversed in the conscious model by atropine and completely prevented from developing in the anaesthetized model that had previously undergone hepatic denervation. Gastric administration of a liquid mixed meal but not glucose or sucrose is capable of activating MIS for purposes of mechanistic studies and quantification of the MIS process. The feeding signal is mediated by the hepatic parasympathetic nerves.

Increased incidence of hepatic insulin-sensitizing substance (HISS)-dependent insulin resistance in female rats prenatally exposed to ethanol.

Insulin causes the release of the hepatic insulin-sensitizing substance (HISS) from the liver. Hepatic parasympathetic nerves play a permissive role in the release of HISS. HISS-dependent insulin resistance (HDIR) occurs in the absence of HISS. Fetal ethanol exposure has been shown to cause dose-dependent HDIR in adult male rat offspring. Since female offspring are more severely affected by in utero ethanol toxicity, we hypothesized that fetal alcohol exposure causes higher incidence and more severe HDIR in adult female offspring. Adult female rat offspring prenatally exposed to different concentrations of ethanol (0%, 15%, and 20%) were tested for insulin sensitivity using the rapid insulin sensitivity test (RIST). The RIST index was significantly reduced in the 15% (134.1 +/- 16.1 mg/kg) and the 20% (98.7 +/- 9.7 mg/kg) group compared with the 0% (220.9 +/- 27.6 mg/kg) group. Administration of atropine produced significant additional HDIR in the 15% group (82.9 +/- 14.5 mg/kg) but not the 20% group (83.8 +/- 20.5 mg/kg) indicating complete HDIR had been produced in this group, contrary to the adult male offspring in a previous study. The results are consistent with the hypothesis that adult-female offspring are more severely affected by in utero ethanol exposure compared with adult-male offspring.

Alcohol suppresses meal-induced insulin sensitization.

BACKGROUND: In the fed state, the glucose disposal action of insulin can be attributed in approximately equal part to the direct action of insulin and to a hepatic insulin sensitizing substance (HISS) that acts selectively on skeletal muscle. HISS action is absent in the 24-hour fasted state. The objective of this study was to determine whether alcohol administered with a meal affected meal-induced insulin sensitization (MIS). METHODS: Rats were fasted for 24 hours and anesthetized, and insulin sensitivity was evaluated using the rapid insulin sensitivity test (RIST). A liquid mixed meal was injected into the stomach along with diluted alcohol equivalent to 1, 0.25, and 0.125 mL/kg of ethanol. After 90 minutes, a second RIST was carried out, atropine administered, and a third RIST done. A control fed group received no ethanol and a RIST was determined at 90 minutes; then 1 mL/kg ethanol or water was administered and the RIST was tested 1 hour later. RESULTS: Ethanol co-administered with a meal produced a dose-dependent suppression of MIS. MIS was blocked back to fasting levels by atropine. MIS developed to a maximal level by 90 minutes and was maintained for at least 1 hour more. Ethanol (1 mL/kg) administered after MIS was fully developed resulted in complete suppression of insulin sensitivity back to a fasting level. CONCLUSIONS: MIS can be demonstrated in response to gastric administration of a liquid mixed meal in anesthetized rats. Alcohol produces a dose-related suppression of MIS and can completely reverse MIS once it has been developed. The concentrations of alcohol reached were well within levels attained by social drinkers. MIS was confirmed to act through HISS action, and neither MIS nor alcohol affected the HISS-independent component of insulin action.

Pharmaceutical reversal of insulin resistance.

Insulin action is approximately doubled following a meal. The mechanism of postprandial insulin sensitization is dependent on hepatic parasympathetic nerves regulated by the prandial status. The nerves provide a permissive signal to the liver that allows insulin to cause the release of a putative hepatic insulin sensitizing substance (HISS) that selectively stimulates glucose uptake into skeletal muscle but not liver or adipose tissue. The parasympathetic signal has several steps identified in the regulatory pathway; acetylcholine acts on muscarinic receptors leading to activation of nitric oxide synthase and generation of HISS. The meal-induced insulin (MIS) sensitization requires hepatic GSH, which decreases with fasting and several disease states. Interfering with the MIS process results in severe insulin resistance with the response to insulin being reduced by approximately 50% to levels seen in the fasted state. A wide range of conditions have been shown to be associated with insulin resistance attributed to lack of the MIS process including insulin resistance; in chronic liver disease produced by chemical damage or bile duct ligation, hepatic denervation, sucrose fed rats, aging, spontaneously hypertensive rats, fetal alcohol exposed adult offspring, spontaneously insulin resistant rats, animals with pharmacological blockade of hepatic muscarinic receptors, NO synthase, cyclooxygenase, hepatic cGMP, and hepatic GSH levels. Pharmaceutical reversal of insulin resistance has been shown in several models using a variety of approaches including mimicking or potentiating the parasympathetic signal using cholinergic agonists, NO donors, cholinesterase antagonists, phosphodiesterase antagonists, and replenishment of hepatic GSH levels. These compounds are being evaluated for therapeutic application by our international academic/industry collaborative team. The MIS process has now been demonstrated in mice, rats, guinea pigs, cats, dogs, and humans, and has been demonstrated by independent laboratories.

A preliminary investigation of the effects of maternal ethanol intake during gestation and lactation on brain adenosine A(1) receptor expression in rat offspring.

Ethanol exposure during fetal development can result in behavioral and neurological deficits, including reduced cognitive functions, retarded growth, and craniofacial abnormalities. Adenosine is an endogenous neuromodulator that fine-tunes the release and/or synaptic activities of several neurotransmitters, including glutamate, dopamine, and serotonin. Our aim was to determine whether ethanol exposure during early development affects adenosine receptors, particularly the A1 receptor subtype, in adult rats. Female rats were given water or 15% (vol/vol) ethanol in water prior to mating and throughout gestation and lactation. Sixty-day-old male rat offspring from these dams were randomly selected and assayed for adenosine A1 receptor expression in four brain areas: cortex, cerebellum, hippocampus, and striatum. Our results indicate that ethanol intake by dams decreased body and brain weights of offspring and reduced both A1 receptor mRNA and protein density in cortex and cerebellum. These preliminary findings indicate that ethanol intake by dams during pregnancy and lactation can affect adenosine A1 receptor signalling in the offspring. A pair-fed controlled study is warranted to explore these findings further.