Lack of ghrelin secretion in response to fasting in cholecystokinin-A (-1), -B (-2) receptor-deficient mice

Cholecystokinin receptors (CCK-Rs) have been classified into two subtypes: CCK-AR (1R) and -BR (2R). We generated CCK-AR(-/-), CCK-BR(-/-), and CCK-AR(-/-)BR(-/-) mice and found that the gastric emptying of a liquid meal was increased in CCK-BR(-/-) and AR(-/-)BR(-/-) mice, compared with wild-type and CCK-AR(-/-) mice. Given that enhanced gastric emptying leads to eating, food intake after overnight fasting was examined, as was the effect of CCK-8S on food intake. Male mice 6-8 months of age were deprived of food for 16 h with free access to water, after which they were injected intraperitoneally (0.1 ml/mouse) with either vehicle or CCK-8 (0.3, 1.0, or 3.0 nmol/mouse), and their food intake was monitored for 4 h. CCK-8S inhibited food intake in wild-type and CCK-BR(-/-) mice, but not in CCK-AR(-/-) or AR(-/-)BR(-/-) mice. Unexpectedly, we observed a lower food intake in CCK-AR(-/-)BR (-/-) mice treated with vehicle than in mice of the other genotypes. To examine the mechanism of decrease in food intake in CCK-AR(-/-)BR(-/-) mice, the involvement of ghrelin was determined in wild-type and CCK-AR(-/-)BR(-/-) mice. Fasting plasma ghrelin levels were significantly lower in CCK-AR (-/-)BR(-/-) mice than in wild-type mice, and no increase in response to fasting was observed in CCK-AR(-/-)BR(-/-) mice. An administration of acyl-ghrelin produced a small increase in food intake in CCK-AR(-/-)BR(-/-) mice, but not to the levels of wild-type mice. In conclusion, CCK-AR(-/-)BR(-/-) mice showed lower food intake as well as lower response to exogenous ghrelin, and a lower plasma ghrelin level after fasting, though which receptor is more important is unknown.     

Electroacupuncture improves restraint stress-induced delay of gastric emptying via central glutaminergic pathways in conscious rats

Acupuncture has been used for treating functional gastrointestinal (GI) disorders. Animal studies demonstrated that acupuncture improves various stress-induced physiological responses. We investigated the effects of electroacupuncture (EA) at ST-36 (Zusanli; lower limb) on stress-induced delay of gastric emptying. Solid food gastric emptying in 90 min was significantly delayed by restraint stress (27.3 +/- 2.1%, n = 8), compared to that of controls (64 +/- 2.1%, n = 8). Restraint stress-induced delay of gastric emptying was significantly restored by the intracisternal (IC)-injection of GABA(A) receptor antagonist, bicuculline methiodide (46.5 +/- 3.1%; n = 6) and GABA(B) receptor antagonist, phaclofen (48 +/- 3.3%; n = 6). Delayed gastric emptying induced by restraint stress was significantly improved by EA at ST-36 (49.7 +/- 1.4%). The stimulatory effect of EA on stress-induced delay of gastric emptying was prevented by pretreatment with IC-injection of glutamate receptor antagonist, kynurenic acid (30.1 +/- 2.1%). In conclusion, restraint stress-induced delay of gastric emptying is mediated via central GABA(A) and GABA(B) receptors. EA at ST-36 stimulates glutaminergic neurons in the brainstem resulting in improvement of stress-induced delay of gastric emptying.     

Repeated stress in young and old 5-HT(2C) receptor knockout mice

Serotonin (5-HT)(2C) receptor null mutant (knockout, KO) mice develop hyperphagia and midlife obesity. Based upon previous observations indicating altered responsiveness to stressful environmental conditions in these mice, we hypothesized that this KO mouse was hyperresponsive to repeated stress. To test this, we examined the effect of two intensities of repeated stress on food intake and body weight in 5-HT(2C) receptor KO and wild-type (WT) mice. The stressors involved daily cage change (including handling) for 3 days then daily restraint for 4 days. On the final day, mice were immediately decapitated after restraint to assess levels of plasma hormones. Two ages were used: young (12 weeks) and old (32-34 weeks). Basally, young KO were prehyperphagic and weighed the same as WT. In the old mice, KO were frankly hyperphagic and heavier than WT. In response to repeated cage change alone, the genotype-specific difference in food intake in the young group was enhanced, whereas in the old group it was diminished. This stressor did not significantly affect body weight change or caloric efficiency with respect to age or genotype. Repeated restraint had little effect on the young mice. However, in the old mice, KO had decreases in relative body weight and caloric efficiency compared with WT. In the old KO mice, adrenocorticotrophic hormone (ACTH), corticosterone and insulin were increased compared with WT mice. Together, these findings indicate that 5-HT(2C) receptor KO mice are hyperresponsive to repeated stress and this effect is influenced by stressor intensity and initial metabolic state of the mouse.     

Overeating,alcohol and sucrose consumption decrease in CB1 receptor deleted mice

Administration of the cannabinoid CB1 receptor antagonist SR141716 (3-10 mg/kg i.p.) abolished neuropeptide Y-induced overeating and significantly reduced ethanol and sucrose intake in CB1 wild-type (+/+) mice. In CB1 receptor knockout (-/-) mice, neuropeptide Y totally lost its capacity to increase food consumption. Similarly, sucrose and ethanol intakes were significantly lower in CB1-/- vs. CB1+/+ mice. In CB1 deficient mice, SR141716 had no effect in these models.     

Intracerebroventricularly administered urocortin inhibits gastric emptying in mice

Urocortin, a recently identified member of the corticotropin-releasing factor (CRF) family, is implicated in the central control of appetite and energy metabolism. We previously reported that peripherally administered urocortin inhibits gastric emptying in conscious mice. In this study, we investigated the effect of intracerebroventricularly administered urocortin on gastric emptying, food intake and body weight in mice. Urocortin decreased food intake and body weight gain more potently than CRF. It significantly decreased gastric emptying of a solid meal; an effect that was inhibited by simultaneous administration of alpha-helical CRF9-41, a CRF antagonist. These results suggest that the potent anorectic properties of urocortin may be partly due to the anti-gastroprokinetic activity of the peptide.     

逆行胃电刺激对健康人水负荷、摄食量和胃排空的影响

目的观察逆行胃电刺激(RGES)对健康人胃容受性、摄食量和胃排空的影响。方法选择健康志愿者12人。沿胃大弯距幽门5 cm处放置一对黏膜电极,以胃动过速的频率9 cyc les/m in,波宽500 m s,强度5 mA的脉冲信号进行RGES。进行水负荷试验、摄食试验和核素闪烁扫描固体胃排空试验来评价RGES的效果。结果与对照阶段相比,RGES可引起摄水量减少13%(P<0.01),摄食减少16%(P<0.001),胃半排空时间延迟33%(P<0.05),120 m in食物存留率增加15%(P<0.05)。水负荷试验、摄食试验和胃排空试验应用的RGES参数没有引起显著症状。结论急性RGES可以显著减少摄水摄食,延缓胃排空,且不引起明显症状。     

Postgastric satiety in the sham-feeding rat

A sham-feeding model using rats fitted with gastric and duodenal cannulas was employed to investigate the role of postgastric mechanisms of satiety in the short-term control of food intake. When fasted rats sham fed a liquid diet [Vivonex High Nitrogen (VHN), 0.5 kcal/ml], food drained freely from gastric fistulas, and mean first-meal size and 90-min intake increased more than threefold. Varying the rate of duodenal infusion of diet during sham feeding (0.06-0.44 kcal/min) decreased first-meal size and total intake in a dose-dependent manner. First meals ended when mean loads of 2-3 kcal had been delivered. The threshold rate (0.11 kcal/min) decreased meal size and total intake by more than 50%. When fasted rats consumed VHN to satiety with closed gastric fistulas, rate of gastric emptying of diet during feeding averaged 0.32 +/- 0.02 kcal/min and the load emptied by meal termination averaged 3.8 +/- 0.2 kcal. These results indicate that rates of gastric emptying of diet and loads delivered to the small intestine following ingestion of liquid food are sufficient to elicit postgastric satiety in the absence of gastric distension.     

Effects of oral glucose intake on gastric myoelectrical activity and gastric emptying.

To investigate the effect of oral glucose intake on gastric motility, we measured gastric myoelectrical activity and gastric emptying on two test conditions: 1) glucose intake and 2) water intake in the same 10 healthy male volunteers (20 to 29 years old). Gastric motility was evaluated with cutaneous-recorded electrogastrography (EGG) for 30 min both on fasting and after glucose or water intake, while gastric emptying was measured using acetaminophen-absorption method. There were no significant changes in EGG dominant frequency after water intake, but the frequency increased significantly after glucose intake. A postprandial dip (i.e., a transient decrease in frequency immediately after the food intake) was observed in 3 subjects after water intake and in 8 subjects following glucose intake. The EGG power ratio was significantly larger after glucose than water intake, with delayed gastric emptying in the former case. These results suggest that glucose is one of the components responsible for postprandial gastric motility.     

Inhibitory effect of the anorexic compound oleoylethanolamide on gastric emptying in control and overweight mice

Gastric emptying regulates food intake. Oleoylethanolamide (OEA), an endogenous acylethanolamide chemically related to the endocannabinoid anandamide, inhibits food intake, but its effect on gastric emptying is unknown. Here, we investigated the effect and the role of OEA on gastric emptying in mice fed either a standard (STD) or a high-fat diet (HFD) for 14 weeks. Gastric emptying was reduced by OEA, but not by its saturated analog, palmitoylethanolamide. The effect of OEA was unaffected by rimonabant (cannabinoid CB₁ receptor antagonist), SR144528 (cannabinoid CB₂ receptor antagonist), 5′-iodoresiniferatoxin (transient receptor potential vanilloid type 1 antagonist), or MK886 (peroxisome proliferator-activated receptor-α) antagonist. Compared to STD mice, HFD mice showed delayed gastric emptying and higher levels of gastric OEA. HFD-induced increase in OEA levels was accompanied by increased expression of the OEA-synthesizing enzyme N-acyl-phosphatidylethanolamine-selective phospholipase D and decreased expression of the OEA-degrading enzyme fatty acid amide hydrolase. These results might suggest that elevation of gastric OEA could possibly contribute to the delayed gastric emptying observed in HFD-fed animals. HFD regulates OEA levels in the stomach through an increase of its biosynthesis and a decrease of its enzymatic degradation. The inhibitory effect of OEA on gastric emptying here observed might underlie part of the anorexic effects of this compound previously reported.

Xenin delays gastric emptying rate and activates the brainstem in mice

Xenin, a 25-amino acid gastrointestinal peptide, inhibits feeding by acting through the central nervous system. Gastrointestinal hormones reduce food intake partly by activating the brainstem and inhibiting gastric emptying. Therefore, we hypothesized that xenin delays gastric emptying through the activation of the brainstem cells. To address this hypothesis, we examined the effect of intraperitoneal (i.p.) injection of xenin on gastric emptying rate and brainstem Fos expression in mice. Gastric emptying rate was reduced by about 93% in xenin-treated mice compared to saline-treated control mice. The i.p. xenin injection significantly increased Fos-immunoreactive cells in the nucleus of the solitary tract (NTS) of the brainstem, but not area postrema (AP) and dorsal motor nucleus of the vagus (DMV). These findings support the hypothesis that xenin-induced anorexia is at least partly due to delayed gastric emptying and the activation of the NTS cells.     

Effects of cholecystokinin-octapeptide (CCK-8) on food intake and gastric emptying in man

Food intake and gastric emptying were measured simultaneously after cholecystokinin-octapeptide (CCK-8) and saline infusions in order to test the hypothesis that reduction in gastric emptying mediates the effect of CCK-8 on food intake. Each of twelve nonobese healthy men received intravenous infusions of CCK-8 and saline on separate nonconsecutive days after they had consumed 500 g of tomato soup tagged with technetium-99-DTPA. Intake of a test meal was measured 20 min after consumption of the soup while gastric emptying was simultaneously monitored by gamma emission scintigraphy of the soup. Food intake and gastric emptying of the soup were both significantly reduced by CCK-8 infusions in comparison to saline. There was a significant correlation between the amount of the test meal eaten and the amount of soup emptied during the period the test meal was being eaten, but not before the meal, only on days when CCK-8 was infused. Differences in intakes between days when saline was infused and days when CCK-8 was infused did not correlate with differences in gastric emptying of soup. These results suggest that CCK may amplify signals of satiety in proportion to the fullness of the stomach. Gastric emptying per se may not mediate the effects of CCK-8 on food intake.     

Amylinergic control of food intake

Amylin is a pancreatic B-cell hormone that plays an important role in the regulation of nutrient fluxes. As such, amylin reduces food intake in laboratory animals and man, slows gastric emptying and it reduces postprandial glucagon secretion. Amylin deficiency which occurs concomitantly to insulin deficiency in diabetes mellitus, may therefore contribute to some of the major derangements associated with this disorder (hyperphagia, excessive glucagon secretion, accelerated rate of gastric emptying). The described actions of amylin all seem to depend on a direct effect of amylin on the area postrema (AP). As to amylin's satiating effect, the physiological relevance of this action is underlined by studies involving specific amylin antagonists and amylin-deficient mice. In the AP, amylin seems to modulate the anorectic signal elicited by CCK. Subsequent to AP activation, the amylin signal is conveyed to the forebrain via distinct relay stations. Within the lateral hypothalamic area, amylin diminishes the expression of orexigenic neuropeptides such as orexin and MCH. Whether these effects contribute to amylin's short term satiating action remains to be determined. Recent studies suggest that amylin may also play a role as a long-term, lipostatic signal, especially when other feedback systems to the brain are deficient. Obese, leptin-resistant Zucker rats which are hyperinsulinemic and hyperamylinemic, were chronically infused with the amylin antagonist AC 187. AC 187 significantly elevated food intake in obese Zucker rats while having no effect in lean controls. This indicates that at least under certain conditions, chronic blockade of endogenous amylin action may lead to an increase in food intake and/or body weight. As mentioned, the site and mechanism of action for peripheral amylin to reduce food intake seems to be well established. It is less clear how centrally administered amylin reduces food intake although it is well known that 3rd ventricular administration of amylin produces a very strong and long-lasting anorectic action. Amylin receptors have been described in various hypothalamic nuclei but the endogenous ligand of these receptors remains to be investigated. The same holds true as to the physiological relevance of the anorectic effect seen after central amylin administration.     

Corticotropin-releasing factor as well as opioid and dopamine are involved in tail-pinch-induced food intake of rats

Several kinds of stress such as psychological stress, restraint, and foot shock inhibit feeding behavior through corticotropin-releasing factor (CRF). In contrast, a mild tail pinch increases food intake in rats. Although dopamine and opioid are thought to be involved in tail-pinch-induced food intake, it is unknown whether CRF participates in this phenomenon. Therefore, we attempted to clarify this issue using rats. A 30-s tail pinch increased food intake in 30 min after the tail pinch, and this increase was blocked by intraperitoneal injection of CRF receptor type 1 selective antagonist. CRF increased food intake in 30 min after intracerebroventricular injection at a dose of 2 or 10 ng, and this increase was also blocked by CRF receptor type 1 antagonist. Tail-pinch- or CRF-induced food intake was blocked by naloxone, pimozide, and spiperone. These results suggest that CRF, through CRF receptor type 1 as well as opioid and dopaminergic systems, are involved in the mechanism of tail-pinch-induced food intake. The results also suggest that brain CRF has dual effects on food intake, hyperphagia and anorexia, in a stress-dependent manner.     

Interactions between gastric emptying and satiety, with special reference to glucagon-like peptide-1.

The slowing of gastric emptying is an important mechanism for the satiating effect of gut peptide signaling. After food intake, cholecystokinin (CCK), as well as glucagon-like peptide-1 (GLP-1) and glucagon-like peptide-2 (GLP-2), are released from the gastrointestinal tract to mediate satiety. In humans, CCK and the GLP-1 have been found to cause satiety in both normal and obese subjects. This satiating effect may be caused by the peptides circulating as hormones with direct effects in the central nervous system, or indirect effects through signals mediated either via the vagus nerve or by activation of vagal afferent fibers due to slow gastric emptying. These peptides also cause gastric relaxation, considered an additional component in the satiating effect of the peptides. To conclude, after food intake, gut peptides may act in concert as neurohormonal satiety signals acting directly in the brain or indirectly via the vagus nerve, as well as through gastric sensory mechanisms to limit food intake.     

Somatostatin Receptor type 2 Mediates Bombesin-Induced Inhibition of Gastric Acid Secretion in Mice

Studies in isolated mouse stomach showed that bombesin releases somatostatin. We characterized the effects of exogenous bombesin on gastric acid secretion in mice and determined the involvement of somatostatin and somatostatin receptor type 2 (SSTR2) by using somatostatin immunoneutralization, the SSTR2 antagonist, PRL-2903, and SSTR2 knockout mice. Gastric acid secretion was monitored under basal and pentagastrin-, histamine- or bethanechol-stimulated conditions in urethane-anaesthetized mice. Bombesin (10–40 μg kg⁻¹ h⁻¹) and somatostatin-14 (20 μg kg⁻¹ h⁻¹) were infused i.v. 10 and 30 min after PRL-2903 or somatostatin antibody pretreatment, respectively. Urethane-anaesthetized wild-type mice had low basal acid secretion (0.12 ± 0.01 μmol (10 min)⁻¹) compared with SSTR2 knockout mice (1.43 ± 0.10 μmol (10 min)⁻¹). Somatostatin antibody and PRL-2903 increased basal secretion in wild-type mice but not in SSTR2 knockout animals. In wild-type mice, bombesin inhibited secretagogue-stimulated acid secretion in a dose-dependent manner, and somatostatin-14 inhibited pentagastrin-stimulated secretion. In wild-type mice pretreated with somatostatin antibody or PRL-2903 and in SSTR2 knockout mice, bombesin and somatostatin-14 i.v. infusion did not alter the increased gastric acid secretion. These results indicate that, in mice, bombesin inhibits gastric acid secretion through the release of somatostatin and the activation of SSTR2. These observations strengthen the important role of SSTR2 in mediating somatostatin inhibitory actions on gastric acid secretion.     

Sustained acceleration of colonic transit following chronic homotypic stress in oxytocin knockout mice

Acute restraint stress delays gastric emptying and accelerates colonic transit via central corticotropin releasing factor (CRF) in rats. In contrast, central oxytocin has anxiolytic effects and attenuates the hypothalamus–pituitary–adrenal (HPA) axis in response to stress. Our recent study showed that up regulated oxytocin expression attenuates hypothalamic CRF expression and restores impaired gastric motility following chronic homotypic stress in mice. We studied the effects of acute and chronic homotypic stress on colonic transit and hypothalamic CRF mRNA expression in wild type (WT) and oxytocin knockout (OXT-KO) mice. Colonic transit was measured following acute restraint stress or chronic homotypic stress (repeated restraint stress for 5 consecutive days). ⁵¹Cr was injected via a catheter into the proximal colon. Ninety minutes after restraint stress loading, the entire colon was removed. The geometric center (GC) was calculated to evaluate colonic transit. Expression of CRF mRNA in the supraoptic nucleus (SON) was measured by real time RT-PCR. Colonic transit was significantly accelerated following acute stress in WT (GC = 8.1 ± 0.8; n = 7) and OXT KO mice (GC =9.4 ± 0.3; n = 7). The accelerated colonic transit was significantly attenuated in WT mice (GC = 6.6 ± 0.5; n = 9) following chronic homotypic stress while it was still accelerated in OXT KO mice (GC = 9.3 ± 0.5; n = 8). The increase in CRF mRNA expression at the SON was much greater in OXT-KO mice, compared to WT mice following chronic homotypic stress. It is suggested that oxytocin plays a pivotal role in mediating the adaptation mechanism following chronic homotypic stress in mice.     

Central oxytocin attenuates augmented gastric postprandial motility induced by restraint stress in rats

Restraint stress delays gastric emptying via uncoordinated motility pattern in rats. Central oxytocin has anxiolytic effects and attenuates the hypothalamic-pituitary-adrenal (HPA) axis in response to stress and facilitates stress-induced delayed gastric emptying. However, the role of central oxytocin in regulating gastric motility remains unknown. Postprandial gastric motility was recorded via a strain-gauge transducer, implanted on the antrum in Sprague–Dawley (SD) rats. To investigate whether central and peripheral oxytocin are involved in gastric motility, oxytocin (10 μg) was administered intracerebroventricularly (icv) and intraperitoneally (ip). Central and peripheral oxytocin administration did not affect the postprandial gastric motility under non-stressed conditions. Restraint stress augmented gastric contractions. Central administration of oxytocin, but not peripheral administration, abolished the augmented postprandial gastric contractions induced by restraint stress. Oxytocin facilitates stress-induced delayed gastric emptying via alleviating uncoordinated gastric motility. Oxytocin might be a candidate for the treatment of stress-induced GI motility disorders.     

Comparison of the effects of a high-fat and high-carbohydrate soup delivered orally and intragastrically on gastric emptying, appetite, and eating behaviour.

To investigate the effects of fat and carbohydrate on appetite, food intake and gastric emptying with and without the influence of orosensory factors, a group of nine healthy, fasted male subjects took part in two separate paired experiments involving high-fat and high-carbohydrate radiolabelled soup preloads. In the first experiment subjects received direct intragastric isocaloric infusions of either a high-fat tomato soup or a high-carbohydrate tomato soup (400 kcal in 425 mL) over 15 min, on two occasions. In the second paired experiment subjects ingested the same high-fat and high-carbohydrate soup over 15 min. In both experiments ratings of hunger and fullness were recorded over a period of 135 min and gastric emptying was measured by scintigraphy. Food intake was evaluated from a test meal (yoghurt drink) given 2 h after the end of the soup infusion/ingestion. When soup was administered intragastrically (Experiment 1) both the high-fat and high-carbohydrate soup preloads suppressed appetite ratings from baseline, but there were no differences in ratings of hunger and fullness, food intake from the test meal, or rate of gastric emptying between the two soup preloads. When the same soups were ingested (Experiment 2), the high-fat soup suppressed hunger, induced fullness, and slowed gastric emptying more than the high-carbohydrate soup and also tended to be more effective at reducing energy intake from the test meal. The results of these studies demonstrate that orosensory stimulation plays an important role in appetite regulation, and also indicate that subtle differences in orosensory stimulation produced by particular nutrients may profoundly influence appetite and gastrointestinal responses.