Insulin resistance induced by sucrose feeding in rats is due to an impairment of the hepatic parasympathetic nerves.

AIMS/HYPOTHESIS: A considerable proportion of whole-body insulin-stimulated glucose uptake is dependent upon the hepatic insulin-sensitising substance (HISS) in a pathway mediated by the hepatic parasympathetic nerves (HPNs). We tested the hypothesis that a high-sucrose diet leads to the impairment of the HPN-dependent component of insulin action. METHODS: We quantified insulin sensitivity using the rapid insulin sensitivity test, a modified euglycaemic clamp. Quantification of the HPN-dependent component was achieved by administration of a muscarinic receptor antagonist (atropine, 3 mg/kg). RESULTS: Insulin sensitivity was higher in standard-fed than in sucrose-fed Wistar rats (305.6+/-34.1 vs 193.9+/-13.7 mg glucose/kg body weight; p<0.005) and Sprague-Dawley rats (196.4+/-5.9 vs 95.5+/-16.3 mg glucose/kg body weight; p<0.01). The HPN-independent component was similar in the two diet groups. Insulin resistance was entirely due to an impairment of the HPN-dependent component in both Wistar rats (164.3+/-28.1 [standard-fed] vs 26.5+/-7.5 [sucrose-fed] mg glucose/kg body weight; p<0.0001) and Sprague-Dawley rats (111.7+/-9.5 vs 35.3+/-21.4 mg glucose/kg body weight; p<0.01). Furthermore, HPN-dependent insulin resistance in Sprague-Dawley rats was already evident after 2 weeks of a high-sucrose diet (28.5+/-7.6 [2 weeks], 35.3+/-21.4 [6 weeks], 17.9+/-5.4 [9 weeks] mg glucose/kg body weight) and was independent of the nature of sucrose supplementation (12.3+/-4.7 [solid] and 17.9+/-5.4 [liquid] mg glucose/kg body weight). CONCLUSIONS/INTERPRETATION: Our results support the hypothesis that insulin resistance caused by sucrose feeding is due to an impairment of the HPN-dependent component of insulin action, leading to a dysfunction of the HISS pathway.

Intrahepatic adenosine triggers a hepatorenal reflex to regulate renal sodium and water excretion.

The mechanism for water and sodium retention in liver cirrhosis is related to the disturbance in hepatic portal circulation. We hypothesize that the increases in intraportal adenosine, which occur when the portal blood flow decreases, may trigger the hepatorenal reflex to inhibit renal water and sodium excretion. In anesthetized rats, intravenous vs. intraportal adenosine-induced effect on renal water and sodium excretion was compared in normal animals and animals with hepatic or renal denervation, and in the presence of an adenosine receptor antagonist. Compared to saline infusion, intraportal adenosine (0.02 mg kg(-1) min(-1) for 1 h) infusion decreased urine flow by 51.3% (11.7 +/- 2.3 vs. 5.7 +/- 0.5 microl min(-1)) for the first 30 min and by 49% (22.8 +/- 5.4 vs. 11.6 +/- 1.5 microl min(-1)) for the second 30-min duration. Urinary sodium excretion was also decreased. Intraportal administration of an adenosine receptor antagonist (8-phenyltheophylline (8-PT), 3 mg kg(-1) bolus injection followed by 0.05 mg kg(-1) min(-1) continuous infusion), as well as liver or kidney denervation, abolished adenosine-induced inhibition. In contrast, intravenous adenosine infusion had no influence on either urine flow or sodium excretion. The data indicated that selectively increased intraportal adenosine inhibited renal water and sodium excretion. The water and sodium retention commonly seen in the hepatorenal syndrome may be related to intraportal adenosine accumulation due to the decrease in intraportal portal flow.

Shear stress-induced nitric oxide release triggers the liver regeneration cascade.

The trigger of the liver regeneration cascade is currently unknown and has been the subject of debate. We hypothesize that, following 2/3 partial hepatectomy (PHX), an increase in the blood flow-to-liver mass ratio results in shear stress-induced nitric oxide (NO) release, which triggers the liver regeneration cascade. Portal venous pressure (PVP), reflecting shear stress in the liver, increased to the same extent following PHX and selective portal vein branch ligation (PVL), a hemodynamic model of PHX, suggesting similar amounts of shear stress in both models. Two indices of the initiation of the liver regeneration cascade were used: proliferative factor (PF) activity in blood 4 h after PHX or PVL and hepatic c-fos mRNA expression 15 min. after PHX or PVL. PF activity and c-fos mRNA expression were increased to similar extents after PHX and PVL, suggesting a similar stimulus in both models. PF activity and c-fos mRNA expression were inhibited by administration of the nitric oxide synthase antagonist, l-NAME, and the NO donor, SIN-1, reversed the inhibition in both models. These results provide support for the hypothesis that a hemodynamic change results in increased shear stress in the liver causing generation of NO, which then triggers the liver regeneration cascade.

Hepatic parasympathetic (HISS) control of insulin sensitivity determined by feeding and fasting.

In response to insulin, a hormone [hepatic insulin sensitizing substance (HISS)] is released from the liver to stimulate glucose uptake in skeletal muscle but not liver or gut. The aim was to characterize dynamic control of HISS action in response to insulin and regulation of release by hepatic parasympathetic nerves. Insulin action was assessed by the rapid insulin sensitivity test, where the index is the glucose required (mg/kg) to maintain euglycemia after a bolus of insulin. Blocking HISS release by interruption of the hepatic parasympathetic nerves by surgical denervation, atropine, or blockade of hepatic nitric oxide synthase produced similar degrees of insulin resistance and revealed a similar dynamic pattern of hormone action that began 3--4 min after, and continued for 9--10 min beyond, insulin action (50 mU/kg). HISS action accounted for 56.5 +/- 3.5% of insulin action at insulin doses from 5 to 100 mU/kg (fed). We also tested the hypothesis that HISS release is controlled by the feed/fast status. Feeding resulted in maximal HISS action, which decreased progressively with the duration of fasting.

Glucose disposal by insulin, but not IGF-1, is dependent on the hepatic parasympathetic nerves.

Insulin-like growth factor-1 (IGF-1) has many insulin-like activities, including stimulation of glucose uptake in skeletal muscle. However, those with diabetes or chronic liver disease are insulin resistant but show a normal hypoglycemic response to IGF-1. We have previously shown that insulin sensitivity depends on a hepatic parasympathetic reflex release of a hormone from the liver. The hypothesis was tested that insulin action, but not IGF-1 action, is dependent on the hepatic parasympathetic reflex. Glucose disposal in response to three doses of IGF-1 (25, 100, 200 microg/kg) was determined in rats. IGF-1 at 200 microg/kg had similar effect on glucose disposal as did 50 mU/kg of insulin. Interruption of the hepatic parasympathetic reflex either by surgical ablation of the anterior nerve plexus or by atropine (1.0 mg/kg) resulted in insulin, but not IGF-1, resistance. Sixteen hours of fasting resulted in insulin, but not IGF-1, resistance. In conclusion, insulin, but not IGF-1, triggers the hepatic parasympathetic dependent release of a putative hepatic insulin sensitizing substance (HISS) that stimulates glucose uptake in skeletal muscle.

Nitric oxide inhibits norepinephrine-induced hepatic vascular responses but potentiates hepatic glucose output.

We previously reported that sympathetic nerve-induced vasoconstriction in the intestine resulted in shear stress induced release of nitric oxide (NO) that led to presynaptic inhibition of transmitter release. In contrast, studies in the liver suggested a postsynaptic inhibition of vascular responses, thus leading to the hypothesis tested here that maintained catecholamine release in the liver would result in maintained metabolic catecholamine action in the face of inhibition of vascular responses. In rats, norepinephrine (NE) induced elevations in arterial glucose content were inhibited by NO synthase antagonism (N(omega)-nitro-L-arginine methyl ester (L-NAME), 10 mg/kg, intraportal) but potentiated by NO donor administration (3-morpholinosydnonimine (SIN-1), 0.2 mg/kg, intraportal). The potentiated effect of SIN-1 was abolished by indomethacin (7.5 mg/kg, intraportal). To confirm the hepatic site of metabolic effect, cats were used so that blood flow and hepatic glucose balance could be determined. SIN-1 potentiated NE-induced glucose output from the liver from 5.0 +/- 0.4 to 7.2 +/- 0.6 mg x min(-1) x kg(-1). The potentiation was blocked by methylene blue, a guanylate cyclase inhibitor. Contrary to the glucose response, L-NAME potentiated but SIN-1 attenuated NE-induced portal vasoconstriction. Thus NO is shown to produce differential modulation of vascular and metabolic effects of NE. Vasoconstriction of the hepatic vasculature is inhibited by NO, whereas the glycogenolytic response to NE is potentiated, responses that are probably mediated by prostaglandin.

Nitric oxide mediates hepatic arterial vascular escape from norepinephrine-induced constriction.

The involvement of nitric oxide (NO) in the vascular escape from norepinephrine (NE)-induced vasoconstriction was investigated in the hepatic arterial vasculature of anesthetized cats. The hepatic artery was perfused by free blood flow or pump-controlled constant-flow, and NE (0.15 and 0.3 microg x kg(-1) x min(-1), respectively) was infused through the portal vein. In the free-flow perfusion model, the NE-induced hepatic vasoconstriction recovered from the maximum point of the constriction, resulting in 36.6 +/- 5. 9% vascular escape. Blockade of NO formation with N(omega)-nitro-L-arginine methyl ester (L-NAME, 2.5 mg/kg ipv) potentiated NE-induced maximum vasoconstriction, and the potentiation was reversed by L-arginine (75 mg/kg ipv). Furthermore, NE-induced vasoconstriction became more stable after L-NAME, resulting in an inhibition of vascular escape (7.5 +/- 3.3%), and the inhibition was reversed by L-arginine (23.0 +/- 6.4%). Similar potentiation of NE-induced vasoconstriction and inhibition of hepatic vascular escape by L-NAME (40.4 +/- 4.3% control vs. 10.2 +/- 3.7% post-L-NAME escape) and the reversal by L-arginine were also observed in the constant-flow perfusion model. The data suggest that NO is the major endogenous mediator involved in the hepatic vascular escape from NE-induced vasoconstriction.

The HISS story overview: a novel hepatic neurohumoral regulation of peripheral insulin sensitivity in health and diabetes.

Data are reviewed that are consistent with the following working hypothesis that proposes a novel mechanism regulating insulin sensitivity, which when nonfunctional, leads to severe insulin resistance. Postprandial elevation in insulin levels activates a hepatic parasympathetic reflex release of a putative hepatic insulin-sensitizing substance (HISS), which activates glucose uptake at skeletal muscle. Insulin causes HISS release in fed but not fasted animals. The reflex is mediated by acetylcholine and involves release of nitric oxide in the liver. Interruption of the release of HISS is achieved by surgical denervation of the anterior hepatic nerve plexus, muscarinic receptor blockade, or nitric oxide synthase antagonism and leads to immediate severe insulin resistance. The nitric oxide donor, SIN-1, reverses L-NAME-induced insulin resistance. Denervation-induced insulin resistance is reversed by intraportal but not intravenous administration of acetylcholine or SIN-1. Liver disease is often associated with insulin resistance; the bile duct ligation model of liver disease results in parasympathetic neuropathy and insulin resistance that is reversed by intraportal acetylcholine. Possible relevance of this HISS-dependent control of insulin action to insulin resistance in diabetes, liver disease, and obesity is discussed.

Blood flow-dependent prostaglandin f(2alpha) regulates intestinal glucose uptake from the blood.

Intestinal glucose uptake (GUi) from blood increased when blood flow (BF) was increased. The increase in BF could elevate shear stress. Therefore, we hypothesize that shear stress-induced release of autacoids mediates the increase in GU(i). A surgically separated segment of small intestine was perfused in situ with the use of an arterial circuit in anesthetized cats. Arterial and portal blood samples were taken simultaneously for assessment of GU(i). Adenosine was used to elevate intestinal BF. The GU(i) increased by 45.0 +/- 18.3 from 25.3 +/- 3.8 micromol. min(-1). 100 g tissue(-1) when the BF increased about four times. It was not a direct effect of adenosine because GU(i) was not altered if the flow was held constant. This increase was blocked by a cyclooxygenase inhibitor, indomethacin, but not by nitric oxide synthase blocker N(G)-nitro-L-arginine methyl ester. Furthermore, prostaglandin F(2alpha) (PGF(2alpha)) but not PGE(2) or PGI(2) reversed the blockade of the increase in GU(i) after indomethacin during elevated blood flow, whereas they had no influence on basal uptake. The results suggest that shear stress-induced release of PGF(2alpha) mediated the increase in GU(i) when blood flow was elevated.

Blockade of hepatic nitric oxide synthase causes insulin resistance.

The hypothesis was tested that insulin sensitivity, previously shown to depend on a functional hepatic parasympathetic reflex, was mediated by hepatic production of nitric oxide (NO). Insulin sensitivity was measured using the rapid insulin sensitivity test. N-nitro-L-arginine methyl ester (L-NAME, 2.5 and 5.0 mg/kg iv) and N-monomethyl-L-arginine (L-NMMA, 0.73 mg/kg), nitric oxide synthase (NOS) antagonists, caused insulin resistance in rats. Intraportal administration of L-NAME at a dose of 1.0 mg/kg significantly reduced the response to insulin (54.9 +/- 5.2%); however, administration of the same dose of L-NAME intravenously did not cause a significant decrease in insulin response. Intraportal, but not intravenous, administration of 3-morpholinosydnonimine (SIN-1, 5. 0 mg/kg), a NO donor, partially reversed the insulin resistance caused by L-NMMA. Intraportal administration of SIN-1 (10.0 mg/kg) completely restored insulin sensitivity after L-NMMA or surgical denervation of the liver. Insulin resistance produced by denervation was not further increased by NOS blockade. These results suggest that blockade of NOS causes peripheral insulin resistance secondary to blockade of the hepatic parasympathetic reflex release of hepatic insulin-sensitizing substance in response to insulin.

Blockade of nitric oxide synthase potentiates the suppression of vasodilators by norepinephrine in the hepatic artery.

We have previously shown that nitric oxide (NO) and adenosine suppress vasoconstriction induced by norepinephrine infusion and sympathetic nerve stimulation in the hepatic artery and superior mesenteric artery. NO is involved in the control of basal vascular tone in the superior mesenteric artery but not the hepatic artery. The vasodilation induced by adenosine is inhibited by NO in the superior mesenteric artery but not in the hepatic artery. Based on these known interactions of catecholamines, adenosine, and NO, the objective of this study was to test the hypothesis that NO modulates the interaction between vasoconstrictors and vasodilators in the hepatic artery. We examined the ability of norepinephrine to suppress adenosine-mediated vasodilation and the role of NO in this interaction. Hepatic arterial blood flow and pressure were monitored in pentobarbital-anesthetized cats. The maximum hepatic arterial vasoconstrictor response to norepinephrine infusion was potentiated by blockade of NO production using Nomega-nitro-L-arginine methyl ester (L-NAME), and the potentiation was reversed by L-arginine. The maximum dilator response to adenosine was only slightly suppressed (14.0+/-5.8%, P < 0.05) by norepinephrine infusion; however, after the NO blockade, the suppression by norepinephrine of the vasodilation induced by adenosine was substantially potentiated (45.2+/-9.1%, P < 0.05). Similar results were obtained for isoproterenol-induced vasodilation. We conclude that the interaction between these vasodilators and norepinephrine was modulated by NO which inhibited the vasoconstriction and the suppression of vasodilators caused by norepinephrine and that in the absence of NO production, norepinephrine-induced constriction and the ability to antagonize dilation is substantially potentiated.

Shear stress-induced nitric oxide antagonizes adenosine effects on intestinal metabolism.

The influence of nitric oxide (NO) on adenosine-induced metabolic effects was studied in the intestine. Blood flow supplied an in situ- isolated segment of small intestine in anesthetized cats via the superior mesenteric artery (SMA) and was controlled by a vascular circuit. The SMA and portal samples were taken for analysis of oxygen and lactate. Adenosine (0.4 mg. kg-1. min-1, intra-SMA) reduced oxygen consumption by 25.1 +/- 2.9 from 73.1 +/- 10.8 micromol. min-1. 100 g-1 and increased lactate production by 13.3 +/- 3.0 from 12.8 +/- 4.6 micromol. min-1. 100 g tissue-1 during constant-flow (CF, decreased shear stress) but not during constant-pressure (CP, increased shear stress) perfusion. Blockade of NO synthase using Nomega-nitro-L-arginine methyl ester did not affect the metabolic effects of adenosine during CF but eliminated the differences seen between CP and CF perfusion. A NO donor, 3-morpholinosydnonimine, attenuated the metabolic effects of adenosine during CF perfusion. The results suggested that shear-induced NO antagonized metabolic effects of adenosine but that the inhibition of vascular effects by NO was not shear dependent since it occurred in both CP and CF perfusion.

Should clinical cardiologists report total peripheral resistance or total peripheral conductance?

Regional and total vascular tone have traditionally been assessed by using calculated resistance as the index. However, in most situations, regional flow or cardiac output changes over a much wider range than does blood pressure. With flow in the denominator (resistance), the index is nonlinearly related to the changing parameter, thus rendering even simple arithmetic means inaccurate. Vascular conductance uses flow in the numerator and results in an index that is linearly related to flow, thus enabling a variety of relationships to be demonstrated that are concealed or distorted when studied by the use of vascular resistance.