Role of sympathetic tone in the loss of syringin-induced plasma glucose lowering action in conscious Wistar rats

Syringin is an active principle purified from the rhizome and root parts of Eleutherococcus senticosus (Araliaceae). The present study is designed to clarify the role of sympathetic activation in the insulinotropic effect of syringin. Plasma glucose lowering effect accompanying with the increase of plasma insulin and C-peptide were obtained in pentobarbital anesthetized Wistar rats 60min after an intravenous (i.v.) injection of syringin (100 microg/kg). However, neither the plasma glucose lowering action, nor the raised plasma levels of insulin and C-peptide can be obtained in conscious rats received same syringin treatment. Otherwise, the insulin-releasing and plasma glucose lowering actions of syringin (100 microg/kg, i.v.) were appeared in conscious rats under chemical sympathectomy using an intraperitoneal injection of guanethidine. In addition, plasma glucose lowering action of syringin (100 microg/kg, i.v.) was observed in conscious rats with alpha1-adrenoceptor blockade by prazosin. The stimulatory actions of syringin on the secretion of plasma insulin and C-peptide were also obtained in prazosin-treated conscious rats. The obtained results suggest that insulinotropic effect of syringin on the plasma glucose regulation is impaired in conscious rats with a regular sympathetic tone; decrease of sympathetic tone as observed in anesthetized animal might be helpful in the therapeutic benefit of syringin.     

Simulation of the autonomic neural control of insulin secretion.

A seventh order, nonlinear, highly isomorphic, dynamic systems model of insulin secretion was used to test specific mechanistic hypotheses about the sites of action of sympathetic and parasympathetic autonomic neural input on glucose-induced insulin secretion. The application of neural input was modeled by changing only those specific model parameters that correspond to the hypotheses under study as a function of the input magnitude. The results of these simulations suggest that sympathetic and parasympathetic inputs modify glucose-induced insulin secretion by decreasing the maximal rate of calcium uptake and increasing the affinity of calcium uptake to glucose, respectively.     

A gene knockout approach in mice to identify glucose sensors controlling glucose homeostasis

A key aspect of glucose homeostasis is the constant monitoring of blood glucose concentrations by specific glucose sensing units. These sensors, via stimulation of hormone secretion and activation of the autonomic nervous system (ANS), regulate tissue glucose uptake, utilization or production. The best described glucose detection system is that of the pancreatic beta-cells which controls insulin secretion. Secretion of other hormones, in particular glucagon, and activation of the ANS, are regulated by glucose through sensing mechanisms which are much less well characterized. Here I review some of the studies we have performed over the recent years on a mouse model of impaired glucose sensing generated by inactivation of the gene for the glucose transporter GLUT2. This transporter catalyzes glucose uptake by pancreatic beta-cells, the first step in the signaling cascade leading to glucose-stimulated insulin secretion. Inactivation of its gene leads to a loss of glucose sensing and impaired insulin secretion. Transgenic reexpression of the transporter in GLUT2/beta-cells restores their normal secretory function and rescues the mice from early death. As GLUT2 is also expressed in other tissues, these mice were then studied for the presence of other physiological defects due to absence of this transporter. These studies led to the identification of extra-pancreatic, GLUT2-dependent, glucose sensors controlling glucagon secretion and glucose utilization by peripheral tissues, in part through a control of the autonomic nervous system.     

Increase of acetylcholine release by Panax ginseng root enhances insulin secretion in Wistar rats

The present study was designed to ascertain the effect of Panax ginseng root on plasma glucose and investigate the possible mechanisms for the effect. Ninety minutes after the oral administration of P. ginseng root to fasting Wistar rats, plasma glucose decreased in a dose-dependent manner. Simultaneous with the reduction in plasma glucose, an increase in the plasma level of insulin and C-peptide was also observed. Moreover, disruption of the available synaptic acetylcholine (ACh), using the inhibitor for choline uptake (hemicholinium-3), or the inhibitor for vesicular choline transport (vesamicol), abolished the metabolic actions of P. ginseng root. Conversely, physostigmine, at a concentration sufficient to inhibit acetylcholinesterase, enhanced the metabolic effect of P. ginseng root. It is possible that P. ginseng root mediates the release of ACh from nerve terminals to enhance insulin secretion. Blockade of the actions of P. ginseng root by 4-diphenylacetoxy-N-methylpiperdine methiodide (4-DAMP) suggested that the site of action is the muscarinic M(3) receptor. Taken together, the results suggest that P. ginseng root has the ability to increase the release of ACh from nerve terminals in rats so as to stimulate muscarinic M(3) receptors activity located in the pancreatic cells for the secretion of insulin, which in turn lower plasma glucose.     

Intracerebroventricular infusion of a triglyceride emulsion leads to both altered insulin secretion and hepatic glucose production in rats

We investigated here whether non-esterified fatty acids (NEFA) influence insulin secretion and action through a direct effect on central nervous system sites involved in the control of glucose homeostasis. Normal Wistar rats received a 48-h intracerebroventricular infusion of either a 10% triglyceride (Intralipid, IL)/heparin emulsion (IL/h) or saline/heparin solution (control). At 48 h, insulin secretion as measured by an intravenous glucose tolerance test, was more elevated in IL/h than in control rats. Pancreatic noradrenaline turnover was decreased by 57% in IL/h rats, suggesting low pancreatic sympathetic output that could account partly for the elevated insulin secretion. The time course of glycaemia was similar in both groups, suggesting insulin resistance. Euglycaemic-hyperinsulinaemic clamps were imposed to assess peripheral and hepatic insulin sensitivity. At each insulin concentration glucose utilization was increased to a similar extent in both groups, whereas hepatic glucose production decreased much less in IL/h than in control rats. Hepatic insulin insensitivity could be related partly to activation of the hypothalamic-pituitary-adrenocortical axis, since plasma corticosterone concentration was significantly increased in IL/h rats compared with controls. Our data indicate that lipids may alter both insulin secretion and hepatic sensitivity to insulin through their effect on central nervous system.     

Reduced sympathetic responsiveness as well as plasma and tissue noradrenaline concentration in growth hormone transgenic mice

AIMS: Acromegaly [overproduction of growth hormone (GH)] and GH deficiency have both been associated with alterations in autonomic nervous system function. The aim of this study was to investigate autonomic nervous system influence on heart rate (HR) in transgenic mice overexpressing bovine GH (bGH). METHODS: HR and HR variability (HRV) were measured in conscious young (8-13 weeks) and old (5-6 months) female bGH and control mice using telemetry. HR control was studied using antagonists and an agonist of adrenergic and muscarinic receptors. Noradrenaline was measured in plasma, heart and kidney using high performance liquid chromatography. RESULTS: Average 24 h resting HR did not differ between bGH and control mice. After saline injection and after muscarinic blockade with methylscopolamine HR increase was blunted (in old) or absent (in young) bGH mice compared with control mice (P < 0.05). Phenylephrine caused a baroreflex mediated decrease in HR from around 550 to 300-350 beats min(-1), not different between bGH and control mice. Time- and frequency-domain measures of HRV were reduced in old bGH compared with control mice (P < 0.05). Noradrenaline concentrations were reduced by 25-49% in plasma and tissue of bGH compared with control mice (P < 0.05). CONCLUSION: The current study suggests reduced autonomic modulation of HR in bGH transgenic mice. Thus, GH appears to have marked effects on autonomic tone, reducing sympathetic nervous system function possibly via reduced noradrenaline stores.     

Whole body response of the peripheral circulation following hemorrhage in the rat.

Changes in mean circulatory filling pressure (MCFP) after hemorrhage reflect the whole-body response of the peripheral circulation to restore the driving force for venous return. In this study, changes in MCFP were measured for 15 min following a rapid 8 ml/kg hemorrhage. Three groups of rats were studied: 1) conscious, untreated; 2) conscious, ganglion blocked; and 3) pentobarbital anesthetized. In all three groups, hemorrhage decreased MCFP approximately 2.6 mmHg immediately after hemorrhage. In the conscious untreated rat, MCFP recovered 1.3 mmHg in 15 min; 83% of this recovery was complete within 2 min, and over 50% was complete by 30 s posthemorrhage. With ganglionic blockade, recovery was slowed to about 70% of that in the conscious, untreated animal during the first 5 min after hemorrhage. MCFP recovery was substantially depressed by pentobarbital, averaging only 42% of that in the untreated animal 5 min after hemorrhage. The results demonstrate that peripheral changes can quickly restore nearly 50% of the MCFP decrease occurring immediately after mild hemorrhage and that about one-third of this response is mediated by the sympathetic nervous system. Pentobarbital anesthesia greatly inhibits recovery, although its repressive mechanism is not known.     

Neural control of insulin secretion: interaction of norepinephrine and acetylcholine

We have quantitatively characterized the interaction of several concentrations of norepinephrine and acetylcholine applied simultaneously on insulin secretion in response to an intermediate glucose concentration (10 mM) from statically incubated pancreatic islets isolated from normal rats. When either norepinephrine or acetylcholine were applied alone, insulin secretion was respectively inhibited or enhanced in a dose-dependent manner. When combinations of norepinephrine and acetylcholine were applied simultaneously to the islet, insulin secretion was enhanced, inhibited, or not affected. The net effects of the interaction of norepinephrine and acetylcholine on insulin secretion were not only a function of the concentration of each neurotransmitter but also the relative concentrations of both neurotransmitters. Analysis of these interaction data suggests that 1) insulin secretion is the result of a complex interaction between sympathetic and parasympathetic neural and metabolic inputs; 2) norepinephrine and acetylcholine modulate insulin secretion primarily by direct action on the beta-cell membrane; 3) insulin secretion is more sensitive to perturbations in norepinephrine concentration than acetylcholine concentrations under these experimental conditions; and 4) the autonomic nervous system has the potential to play a major role in the control of insulin secretion in response to glucose.     

The modulation of visceral functions by somatic afferent activity

We began by briefly reviewing the historical background of neurophysiological studies of the somato-autonomic reflexes and then discussed recent studies on somatic-visceral reflexes in combination with autonomic efferent nerve activity and effector organ responses. Most of the studies that have advanced our knowledge in this area have been carried out on anesthetized animals, thus eliminating emotional factors. We would like to emphasize again that the functions of many, or perhaps all visceral organs can be modulated by somato-sympathetic or somato-parasympathetic reflex activity induced by a appropriate somatic afferent stimulation in anesthetized animals. As mentioned previously, some autonomic nervous outflow, e.g. the adrenal sympathetic nerve activity, is involved in the control of hormonal secretion. John F. Fulton wrote in his famous textbook "Physiology of the Nervous System" (1949) that the posterior pituitary neurosecretion system (i.e. for oxytocin and vasopressin) could be considered a part of the parasympathetic nervous system. In the study of body homeostasis and environmental adaptation it would seem very important to further analyze the contribution of somatic afferent input to the autonomic nervous and hormonal regulation of visceral organ activity. Also, some immunological functions have been found to be influenced by autonomic nerves or hormones (e.g. adrenal cortical hormone and catecholamines). Finally, we must take into account, as we have briefly discussed, that visceral functions can be modulated by somatic afferent input via various degrees of integration of autonomic nerves, hormones, and immunological processes. We trust that such research will be expanded to higher species of mammals, and that ultimately this knowledge of somato-visceral reflexes obtained in the physiological laboratory will become clinically useful in influencing visceral functions.     

A definition of internal constancy and homeostasis in the context of non-equilibrium thermodynamics

The constancy of the internal environment, internal homeostasis, and its stability are necessary conditions for the survival of a biological system within its environment. These have never been clearly defined. For this purpose nonequilibrium thermodynamics is taken as a reference, and the essential principles of equilibrium, reversibility, stationary steady state and stability (Lyapounov, asymptotic, local and global), are briefly illustrated. On this basis, internal homeostasis describes a stationary state of nonequilibrium, the actual state of rest, X(t), resulting from the relation X(t) = Xs + x(t), between a time-independent steady state of reference (Xs), and time-dependent fluctuations of the state variables, x(t). In humans, two resting spontaneous homeostatic states are: (1) the conscious state of quiet wakefulness, during which time-dependent variables display bounded oscillations around the mean time-independent steady state level, this conscious state being thus stable in the sense of Lyapounov, and (2) the unconscious stable state of non-rapid eye movement sleep, in which the time-dependent variables would approach the lowest spontaneously attainable time-independent state asymptotically, sleep becoming a globally stable and attractive state. Exercise may be described as a non-resting, unstable active state far away from equilibrium and hibernation is a resting, time-independent steady state very near equilibrium. The range between sleep and exercise is neurohumorally regulated. For spontaneously stable states to occur, slowing of the metabolic rate, withdrawal of the sympathetic drive and reinforcement of the vagal tone to the heart and circulation are required, thus confirming that the parasympathetic division of the autonomic nervous system is the main controller of homeostasis.     

Efferent sympathetic nervous control of rectal motility in the cat

The sympathetic nervous control of rectal motility was studied in anesthetized cats. Division of the sympathetic nerves, i.e. the hypogastric nerves and the lumbar colonic nerves and alpha-adrenergic blockade reduced rectal tone indicating that these nerves are tonically active. Efferent electrical stimulation of the nerves at high intensities caused an immediate and sustained contraction which was inhibited after phentolamine but unaffected by hexamethonium suggesting a direct alpha-adrenergic effect on the rectal smooth muscle. However when prevailing rectal tone was high beta-adrenergic inhibitory responses unaffected by hexamethonium were observed. In addition the hypogastric nerves seem to convey cholinergic excitatory fibres to the rectum. The results imply that the sympathetic nerves are integrated in the nervous regulation of rectal motility in a fashion similar to the nervous control of the internal anal sphincter.     

Metabolic adaptation of mice in a cool environment

Homeothermic animals, including humans, live by adapting to changes in ambient temperature. Numerous studies have demonstrated cold exposure (at approximately 5°C) improves glucose tolerance despite reducing insulin secretion and increasing energy expenditure. To determine the effects of a small reduction in ambient temperature on energy metabolism, we compared two groups of mice; one exposed to a cool environment (20°C) and the other maintained in a near-thermoneutral environment (25°C) for 10 days. Both glucose-induced insulin secretion and glucose response were significantly impaired in mice exposed to a cool environment. In the cool temperature-exposed mice, skin temperatures were reduced, and plasma norepinephrine levels were increased, suggesting that impairment of insulin secretion was facilitated by induction of sympathetic nervous activity due to skin cooling. In addition, expression of GLUT4 mRNA was increased significantly in inguinal subcutaneous adipose tissue (IWAT) but not in epididymal or brown adipose tissue or skeletal muscle in these mice. Moreover, expression of Dok1, a molecule linked to activation of insulin receptors in adipocyte hypertrophy, and Cd36, a molecule related to NEFA uptake, were also increased at mRNA and/or protein levels only in IWAT of the cool temperature-exposed mice. Fatty acid synthesis was also facilitated, and fat weights were increased only in IWAT from mice kept at 20°C. These results suggest that a small reduction in ambient temperature can affect glucose homeostasis through regulation of insulin secretion and preferentially enhances fat storage in IWAT. These adaptations can be interpreted as preparation for a further reduction in ambient temperature.     

Heart Rate Variability in Conscious and Anesthetized Rats under the Action of Angiotensin Converting Enzyme Inhibitors

We studied the effects of three various angiotensin converting enzyme inhibitors (enalapril, lisinopril and quinapril) on heart rhythm variability in anesthetized and immobilized rats. In all cases (except for quinapril in experiments on anesthetized animals), the preparations reduced the total rhythm variability and, according to spectrum analysis, increased activity of the parasympathetic autonomic nervous system to different degrees and decreased sympathetic tone. Quinapril and lisinopril produced the most pronounced influence on heart rhythm in anesthetized rats; enalapril was less potent in this respect. In immobilized animals, quinapril and enalapril showed the greatest activity and lisinopril the lowest. The more pronounced effect of quinapril both under anesthesia and during immobilization appears to be linked to the highest affi nity of quinaprilat to circulatory and tissue compartments of the renin-angiotensinaldosterone system.     

beta-adrenergic mechanisms modulate central nervous system effects of prolactin on milk ejection

It is known that prolactin (PRL) is produced within the brain and numerous central actions of the hormone have been reported. In anesthetized lactating rats, central administration of PRL, i.e., intracerebroventricular (icv) or intrathecally (it), facilitated milk ejection (ME) by depressing the sympathetically mediated facilitatory tone of the mammary ductal system. However, it is not known whether or not the same effects and similar mechanisms take place in conscious rats after PRL administration. In the present study, the effects of centrally administered PRL, i.e., icv or it, on ME was determined in both conscious and anesthetized rats. In conscious rats, the rate of ME was determined by applying a 15-min period of suckling by the litter, following a 6-h period of isolation. In anesthetized rats, intramammary pressure (IMP) responses of the mammary glands to exogenous oxytocin (OT) were recorded. The results showed that, whereas in anesthetized rats, increased responsiveness of the mammary glands to OT were observed after PRL administration, an intense inhibition of ME occurred in conscious rats. Because, in conscious and anesthetized rats, these effects were prevented by prior administration of the beta-adrenergic blocker propranolol (PROP) to the mothers, this suggests that the PRL effects on ME are modulated through sympathomimetic and sympatholytic actions in conscious and anesthetized rats, respectively. Thus, as shown by ductal tone measurements, in conscious, but not in anesthetized rats, the effect of PRL was associated with increased ductal constriction within the mammary glands; an effect that was mimicked by icv administration of the beta-adrenergic agonist isoproterenol (ISOP) and that was prevented by PROP. Further, the sympatholytic action of icv-PRL in anesthetized rats prevented the effect on ductal tone of both icv-PRL in conscious rats and of ISOP in anesthetized rats. Taken together, these results clearly suggest that the central effects of PRL on ME are modulated by adrenergic mechanisms.     

Tolerance is overcome in beef insulin-transgenic mice by activation of low-affinity autoreactive T cells

To gain insight into the factors controlling the maintenance or loss of T cell self tolerance we produced beef insulin (BI)-transgenic BALB/c mice. Transgenic mice express BI under control of the human insulin promoter and secrete physiological amounts of beef insulin. Although these mice are tolerant to BI, as evidenced by the lack of insulin-specific IgG antibody production following intraperitoneal immunization, tolerance is not complete. Footpad immunization results in a weak antigen-specific T cell proliferative response, indicating the presence of self- reactive BI-specific T cells in the periphery. These T cells are functional in vivo, providing support for IgG1, IgG2a, and IgG2b BI-specific antibody production, but require higher concentrations of antigen than nontransgenic T cells (both in vivo and following recall responses in vitro) to become activated. In vitro, BI-specific T cell proliferation in BI-transgenic mice can be largely restored by addition of interleukin-2, indicating that a significant component of T cell tolerance is mediated by anergy. To characterize the autoreactive T cells that become activated when tolerance is broken, BI-specific T cell hybridomas were generated from transgenic mice and compared to a panel of hybridomas previously derived from nontransgenic BALB/c mice. The majority of BI-transgenic hybridomas recognized the immunodominant A1–14 beef insulin peptide but with lower avidity than BALB/c hybridomas. Consistent with this, none of the dominant T cell receptor rearrangements found in the BALB/c BI-specific T cell receptor repertoire were found in the transgenic hybridomas. These results indicate that, despite evidence for clonal inactivation of many BI-specific T cells in BI-transgenic mice, loss of tolerance results from activation of low-affinity antigen-specific T cells that appear to have escaped this process.     

Divergent neuroendocrine responses to localized and systemic inflammation

The sympathetic nervous system (SNS) is part of an integrative network that functions to restore homeostasis following injury and infection. The SNS can provide negative feedback control over inflammation through the secretion of catecholamines from postganglionic sympathetic neurons and adrenal chromaffin cells (ACCs). Central autonomic structures receive information regarding the inflammatory status of the body and reflexively modulate SNS activity. However, inflammation and infection can also directly regulate SNS function by peripheral actions on postganglionic cells. The present review discusses how inflammation activates autonomic reflex pathways and compares the effect of localized and systemic inflammation on ACCs and postganglionic sympathetic neurons. Systemic inflammation significantly enhanced catecholamine secretion through an increase in Ca²⁺ release from the endoplasmic reticulum. In contrast, acute and chronic GI inflammation reduced voltage-gated Ca²⁺ current. Thus it appears that the mechanisms underlying the effects of peripheral and systemic inflammation neuroendocrine function converge on the modulation of intracellular Ca²⁺ signaling.     

Reflex insulin secretion due to central hyperglycemia promoted by intracarotid glucose infusion in dogs

The reflex insulin secretion was investigated by glucose infusions (10 ml of 3% solutions) into the carotid artery or the jugular vein in anesthetized and unanesthetized, untrained dogs. Neither intrajugular nor intracarotideal glucose infusions changed the peripheral glycemic level in any of the experiments. Reflex insulin secretion did occur when glucose was injected into the carotid artery under pentobarbital anesthesia (30 mg/kg IV), but not in unanesthetized animals. Saline infusion into the carotid artery or glucose into the jugular vein in both groups did not elicit a significant increase in insulin secretion.     

Sympathetic nerve activity in metabolic control – some basic concepts

A role for the sympathetic nervous system in hypertension has been looked for in relation to the ‘metabolic syndrome’ with associations between body weight, insulin sensitivity and hypertension. By use of microneurography human sympathetic responses to hypoglycaemia, normoglycaemic hyperinsulinaemia and food intake have been studied. A strong but differentiated influence of insulin‐induced hypoglycaemia comprises increase in muscle sympathetic nerve activity (MSNA) and the sudomotor part of skin sympathetic nerve activity (SSNA), whereas vasoconstrictor SSNA is inhibited. Responses to infusion of 2‐deoxy‐d ‐glucose are identical, suggesting central nervous system glucopenia and not insulin to be the causative factor. Insulin infusion during normoglycaemia evokes a moderate increase in MSNA; SSNA and blood pressure does not change. After glucose ingestion MSNA displays a sustained increase, which is only partly elicited by insulin. A significant albeit weaker increase occurs after pure protein or fat meals, and after glucose ingestion in C‐peptide‐negative diabetic patients, with no insulin secretion. In healthy elderly people the MSNA response to food intake is weak, because of a high outflow already at rest; this is suggested to explain postprandial hypotension in the elderly, a paradoxical mechanism behind clinical autonomic failure. A pathophysiological role of MSNA in the metabolic syndrome with hypertension has been speculated. An association between obesity and elevated level of MSNA at rest is established; observed relationships to chronic insulin levels and hypertension are less unanimous. The adipous tissue regulating hormone leptin has become one focus of interest in ongoing attempts to elucidate a possible role of the human sympathetic nervous system in the ‘metabolic syndrome’ and hypertension.     

Autonomic control of pacemaker activity in the atrioventricular junction of the dog

A stable atrioventricular (AV) junctional rhythm was induced in open-chest, anesthetized dogs by injecting pentobarbital into the sinus node artery. A factorial experimental design was used to quantify the changes in AV junctional rate as a function of the frequency of cardiac sympathetic and parasympathetic stimulation. The AV junctional pacemaker cells were more responsive to autonomic neural stimulation, but the vagal-sympathetic interactions were less pronounced than had previously been observed for the SA nodal pacemaker cells. In a group of seven animals, sympathetic stimulation at a frequency of 1.4 Hz increased the AV junctional rate by 102% from a control rate of 54 beats/min. In the same animals, vagal stimulation at a frequency of 8.4 Hz reduced the AV junctional rate by 56%. In three other animals, the AV junction was even more responsive; equivalent chronotropic effects were achieved with stimulation frequencies that were only about one-third of those cited above. There was a moderate, but significant, autonomic interaction: in the group of seven animals, the positive chronotropic effect of sympathetic stimulation at 1.4 Hz was 72% greater at the low level (0 Hz) than at the high level (8.4 Hz) of vagal activity.     

Direct tonic inhibition of insulin secretion by central nervous system

An experimental animal model was developed to investigate the influence of the central nervous system (CNS) on insulin secretion via direct innervation to the pancreas. This model eliminates any indirect CNS effect on the pancreas by hormonal factors. It consists of a vascularly isolated, in situ perfused rat pancreas preparation with the cephalic portion of the animal functional, including innervation to the pancreas. Using this rat model, we have demonstrated that the nonstimulated rat brain exerts a tonic inhibition of glucose-stimulated insulin secretion in nonfasted rats. This inhibition can only occur directly via neurons from brain to pancreas, probably via the sympathetic nervous system. The brain does not alter the biphasic pattern of insulin secretion, but it suppresses the overall insulin secretory process throughout both phases of insulin secretion by a relatively constant degree ranging from 38 to 47%. The physiological purpose of this tonic suppression of insulin secretion is open to speculation, but it is well known that tonic sympathetic stimulation of the cardiovascular system represents an important physiological control mechanism. An analogous control may exist with respect to insulin secretion.