Gut hormones are implicated in the regulation of energy balance. The studies in this thesis have examined the effects exercise on gut hormones (acylated ghrelin and peptide YY3-36), appetite and food intake, over extended durations. Sixty-nine young, healthy, predominantly Caucasian males were recruited to six studies. The age, height and body mass of the participants were: 22.4 ± 0.3 y, 1.80 ± 0.1 m, 76.2 ± 1.0 kg (mean ± SEM). In study one, 90 min of resistance exercise did not influence appetite or energy intake over 24 h of assessment, yet stimulated a latent preference for carbohydrate rich foods. Study two demonstrated that appetite was suppressed during 60 min of swimming but was elevated after consuming a post-exercise meal. Plasma acylated ghrelin was suppressed during swimming but was unaltered after. Energy/macronutrient intake remained unchanged. In study three, 60 min of brisk walking (45 ± 2% of max) did not influence appetite, energy/macronutrient intake or plasma concentrations of acylated ghrelin during an eight hour observation period. Study four showed that 90 min of treadmill running (69 ± 1% of max) transiently suppressed appetite and acylated ghrelin but did not influence these variables, or energy/macronutrient intake within 22.5 h after exercise. The findings of study five suggest that the suppression and subsequent rebound in plasma acylated ghrelin after exercise may be related to a delayed voluntary decision to eat after. Finally, study six showed that appetite, food intake and circulating concentrations of acylated ghrelin and peptide YY3-36 are responsive to acute deficits in energy induced by food restriction but are not sensitive to equivalent energy deficits induced by exercise. This thesis has shown that exercise transiently alters circulating levels of acylated ghrelin and peptide YY3-36 in directions expected to inhibit appetite however no changes are seen after exercise. Conversely, food restriction elicits marked compensatory changes in circulating acylated ghrelin and peptide YY3-36. This thesis also demonstrates that resistance exercise, brisk walking and running do not stimulate appetite or energy intake over defined periods, even when the energy expenditure elicited is high. Swimming appears to increase appetite in the latter hours after exercise.

The role of gut hormones in the regulation of appetite and food intake is well established. The studies presented within this thesis have examined the effects of exercise and different environmental conditions on gut hormones (acylated ghrelin and total peptide YY), appetite and food intake. Forty-two young (mean ± SEM; 22.6 ± 0.4 y), healthy and generally lean (body mass index 23.7 ± 0.3 kg m2) males were recruited into four studies. In study one, 60 minutes of high intensity (70 % of O2 max) running and cycling exercise suppressed concentrations of the appetite-stimulating hormone acylated ghrelin to a similar extent. Study two revealed that after 60 minutes running in the heat (30 °C), hunger is lower in the pre-prandial period, and energy intake lower over the 7 h trial duration compared with a similar trial conducted in temperate (20 °C) conditions. Acylated ghrelin was suppressed during running in the temperate and hot environment but this did not appear to mediate the lower energy intake observed during the hot trial. In study three, energy intake tended to be higher after 60 minutes running in a cool environment (10 °C) compared with a temperate (20 °C) environment. During and shortly after running in the cold, perceived ratings of fullness and satisfaction were lower. Acylated ghrelin concentrations appeared to be suppressed to a lesser extent during running in the cold which could mediate the elevated energy intake observed at the first meal. However, energy intake was also higher at the second meal in the cold trial when acylated ghrelin concentrations were higher in the temperate trial. Study four showed that energy intake and acylated ghrelin concentrations were lower, and total PYY tended to be lower, in normobaric hypoxia suggesting a possible role for acylated ghrelin, but not PYY, in mediating the decrease in energy intake observed in hypoxia. This thesis confirms that exercise transiently suppresses acylated ghrelin concentrations regardless of the environmental conditions (temperature and altitude) exercise is performed in. The findings support anecdotal reports that appetite and energy intake are suppressed in the heat and stimulated in the cold. These responses may be partly mediated by acylated ghrelin immediately after running but other mechanisms are likely involved thereafter. Acute hypoxic exposure suppresses acylated ghrelin concentrations; an observation which may explain the decreased energy intake in hypoxia.

L’intestin est un organe majeur de l’organisme de par ses fonctions et sa localisation, établissant une barrière active avec un environnement complexe composé du microbiote intestinal, des aliments digérés et d’éléments sécrétés par l’hôte. Outre ses fonctions digestives, absorptives et immuno-modulatrices, l’intestin est également un important organe endocrinien, sécrétant une vingtaine d’hormones régulant des fonctions physiologiques telles que la prise alimentaire, le métabolisme énergétique ou la digestion et le transit intestinal. Ces hormones sont produites par une famille de cellules épithéliales, les cellules entéroendocrines, et sécrétées en réponse à l’activation de récepteurs reconnaissant des éléments du contenu intestinal. En particulier, les cellules entéroendocrines de type L sécrètent GLP-1 et Peptide YY (PYY), impliqués respectivement dans le contrôle de la sécrétion d’insuline et dans la régulation de la prise alimentaire ainsi que le contôle du transit intestinal. Elles sont majoritairement localisées dans l’iléon et le côlon, là où le microbiote intestinal est le plus dense. Le microbiote intestinal permet notamment la fermentation des fibres en acides gras à chaîne courte (AGCC), la production de vitamines, la maturation du système immunitaire de l’hôte et joue lui-même un rôle de barrière contre les pathogènes. Un dialogue entre le microbiote intestinal et l’hôte est nécessaire dans le maintien de l’homéostasie intestinale, nécessitant la reconnaissance par l’hôte de produits bactériens. En particulier, les récepteurs Toll-Like (TLR) permettent la reconnaissance de motifs moléculaires microbiens conservés et sont impliqués dans l’immunité innée de l’hôte. Certains produits bactériens ont également un rôle physiologique tels que les AGCC qui sont une source d’énergie importante pour les colonocytes, en plus d’activer des voies de signalisation. Il a été montré que des régimes riches en fibres, et donc permettant une production accrue d’AGCC, ou plus directement l’administration d’AGCC dans le colon, induit chez l’Homme ou la souris une augmentation des concentrations plasmatiques de PYY, par des mécanismes encore peu compris. En utilisant des lignées cellulaires humaines modèles de cellules entéroendocrines, nous avons caractérisé les effets des AGCC et des motifs bactériens reconnus par les TLR sur l’expression et la sécrétion de PYY et les réponses calciques dans ces cellules. Nous avons pu démontrer que les TLR sont exprimés de manière fonctionnelle, à l’exception de TLR4 et TLR8 dans ces cellules, et que le butyrate augmente leur expression et leur activité. De plus, la stimulation des TLR augmente l’expression de Pyy d’un rapport de 2, mais a peu d’effet sur la sécrétion dans ces cellules. Les AGCC ont des effets divers sur l’expression et la sécrétion de PYY. Alors que le butyrate et le propionate augmentent très fortement l’expression de Pyy, par des rapports respectivement de 120 et 40, par un mécanisme d’inhibition des déacétylases d’histone et de lysine, l’acétate augmente l’expression de Pyy plus modestement par l’activation des récepteurs aux AGCC FFAR2 et FFAR3. L’activation de FFAR2 par les AGCC induit une forte réponse calcique oscillatoire induisant la sécrétion de PYY alors que l’activation de FFAR3 et de GPR109a par le butyrate diminue la concentration calcique cellulaire et réduit les réponses sécrétoires. Ainsi, les AGCC augmentent la production de PYY et régulent sa sécrétion, mais avec et par des effets différents. Ces travaux ont permis de montrer le rôle des cellules entéroendocrines humaines de type L dans la reconnaissance de produits bactériens par l’expression de TLR et par leurs réponses aux AGCCs modulant l’expression et de la sécrétion de PYY. De plus, ces résultats ont déterminés en partie les mécanismes impliqués dans la réponse bénéfique de l’hôte à la consommation de fibres et l’augmentation de la production d’AGCC. / The human gut exerts major functions, mainly due to its localization and by forming an active barrier between a complex environment made of the gut microbiota, digested food products and secreted elements by the host. The main functions of the gut are digestion and absorption of nutrients and it is the first pool of immune cells and a barrier against pathogens, but the gut is also a main endocrine organ secreting more than twenty different hormones. These hormones regulate a wide range physiological functions including food intake, energy metabolism or digestion. Enteroendocrine cells, a sparse family of intestinal epithelial cells, produce and secrete these hormones in response to the activation of a variety of receptors that sense luminal content. Among them, L-cells secrete GLP-1 and Peptide YY (PYY) that are implicated in the regulation of insulin secretion, food intake and intestinal motility. They are mainly found in the distal ileum and in the colon where the microbiota is the densest. Gut microbiota ferments fibers into short chain fatty acids (SCFAs), produces vitamins, participates in regulation of host immune system and is a barrier against pathogens. The cross talk between microbiota and intestinal epithelium is important to maintain the local homeostasis, and is mediated by host receptors recognizing microbial products. Among them, Toll-like receptors (TLRs) recognize conserved microbial associate molecular patterns (MAMPs) and participate to the host innate immunity. Some microbial products also have important functions for the host such has SCFAs that are an important energy substrate for colonocytes and can also activate different signaling pathways. It was shown that fiber-rich diets, increasing production of SCFAs, as well as direct administration of SCFAs in the colon in humans or mice increased PYY plasma levels through mechanisms still undeciphered. Taking advantage of human cell lines as L-cell models, we assessed the different effects of SCFAs and TLR stimulation on PYY expression and secretion and calcium signaling in these cells. We showed that TLRs are functionally expressed in these cells at the exception of TLR4 and TLR8, and that butyrate, one of the three main SCFAs produced by the microbiota increases cell sensitivity to TLR stimulation by increasing their expression. Moreover, TLR stimulation increases Pyy expression by a fold of two but has little effect on secretion. SCFAs differently regulate Pyy expression. Propionate and butyrate highly increase Pyy expression by a fold of 40 and 120 respectively, and their effects are mainly mediated by inhibition of lysine/histone deacetylases whereas acetate increases expression of Pyy by a fold of 1.8 through stimulation of FFAR2 and FFAR3. SCFAs also induce a strong FFAR2-dependent oscillatory response monitoring PYY secretion whereas butyrate via FFAR3 and GPR109a decreases cytosolic calcium concentration and consequently reduces secretory responses. Thus, SCFAs differently increase PYY production and secretion depending of their chain length. Altogether, these results highlight the role human L-cells in microbiota-host crosstalk by sensing microbial products through expression of TLRs and their responses to SCFAs modulating PYY production and secretion. Furthermore, we deciphered some of the mechanisms implicated in beneficial host response to enriched fiber diets and increased production of SCFAs.

In recent years there has been growing interest in the role of gut hormones in regulating appetite, energy balance and weight control. Prominent among these hormones is the hunger hormone ghrelin which is the only circulating hormone currently known to stimulate appetite. A variety of hormones are known to suppress appetite and notable among these is peptide YY (PYY). Both ghrelin and PYY exist in more than one form with acylated ghrelin and PYY3-36 representing the biologically active forms of these hormones i.e. the form of each hormone with the most potent effects on appetite. Many studies have investigated ghrelin responses to exercise in male participants and some studies have also examined PYY responses. Far fewer studies have examined ghrelin and PYY responses in female participants and this was the primary purpose of the studies reported here. This thesis comprises four main experimental chapters which collectively sought to clarify whether there is any evidence to support the hypothesis that appetite, gut hormone and food intake responses differ in female compared with male participants. A total of 123 participants took part in the studies reported in this thesis. The first of these studies was cross-sectional in nature and compared fasting appetite, plasma acylated ghrelin and dietary restraint questionnaire values (among other variables) in 34 males and 33 females. No significant differences were observed between sexes for any of these variables. In the second study, appetite, plasma acylated ghrelin and ad libitum food intake responses to cycling exercise were examined in 13 female participants taking the oral contraceptive pill in both the luteal and follicular phases of the menstrual cycle. Although fasting hunger and prospective food consumption values were higher in the follicular than the luteal phase there was no difference in appetite, plasma acylated ghrelin and food intake responses to exercise between menstrual cycle phases. In the third study, appetite, plasma acylated ghrelin, plasma PYY3-36 and food intake responses to energy deficits created via diet and exercise were compared in 13 young, healthy female participants who completed three separate trials (control, exercise deficit and food deficit) in a random order. The findings revealed that, as with male participants, females experience compensatory appetite, gut hormone and food intake responses to dietary induced energy deficits but not to exercise induced energy deficits (over the course of a nine hour observation period). The final study reported in this thesis compared appetite, plasma acylated ghrelin and ad libitum food intake responses to a one hour run in 10 male and 10 female participants. Suppressions of both hunger and plasma acylated ghrelin were noted during exercise but there was no significant difference in the responses of males and females during or after exercise. Collectively, the studies reported here suggest: 1) that fasting appetite and plasma acylated ghrelin concentrations do not differ between male and female participants; 2) that appetite, ghrelin and food intake responses to cycling exercise do not differ according to the phase of the menstrual cycle in females; 3) that dietary restriction is more likely to elicit compensatory feeding responses than elevated exercise levels in females and 4) that males and females do not differ in their acute appetite, ghrelin and food intake responses to an acute bout of running exercise. Hence the studies reported here do not support the hypothesis that exercise will be less effective for controlling appetite and food intake in females than in males.

Being able to effectively match energy intake to energy expenditure (EE) is an important aspect in preventing weight re-gain in the post-obese. Although it is generally agreed upon that resting EE decreases concomitantly with weight loss, there is no set standard comparing the deviations with differing weight loss protocols and additionally, controversy remains as to whether this decrease is greater than can predicted. In order to address these issues 2977 subjects were analyzed using a systematic review and the differences of both the protocol and length of various interventions in addition to sex were compared. Next, data was selected from this systematic review and 815 subjects were analyzed for weight loss-induced changes in resting EE, FM and FFM. Another subgroup of studies (n = 1450) was analyzed and compared against the Harris-Benedict prediction equation to determine whether the changes in resting EE were greater than what was expected. Finally, in order to determine which factors may be involved in regulating changes in resting EE during weight loss, a secondary analysis was performed on 28 post-menopausal women (age= 50.4 ± 2.0 yrs; BMI= 32.4 ± 5.2 kg/m²) who were submitted to a 6-month caloric restriction. Body composition (DXA), resting EE (indirect calorimetry), physical activity EE (PAEE) and total EE (TEE) (doubly-labelled water) were measured before and after the 6 month weight loss. Blood samples were collected before and after to measure leptin and peptide YY. The results indicate that there was indeed a depression in resting EE during weight loss regardless of the type of intervention utilized. Furthermore, these findings suggest that the changes could not fully be explained by changes of FM and FFM alone and that leptin may be an important contributor to the changes of resting EE during weight loss.

Being able to effectively match energy intake to energy expenditure (EE) is an important aspect in preventing weight re-gain in the post-obese. Although it is generally agreed upon that resting EE decreases concomitantly with weight loss, there is no set standard comparing the deviations with differing weight loss protocols and additionally, controversy remains as to whether this decrease is greater than can predicted. In order to address these issues 2977 subjects were analyzed using a systematic review and the differences of both the protocol and length of various interventions in addition to sex were compared. Next, data was selected from this systematic review and 815 subjects were analyzed for weight loss-induced changes in resting EE, FM and FFM. Another subgroup of studies (n = 1450) was analyzed and compared against the Harris-Benedict prediction equation to determine whether the changes in resting EE were greater than what was expected. Finally, in order to determine which factors may be involved in regulating changes in resting EE during weight loss, a secondary analysis was performed on 28 post-menopausal women (age= 50.4 ± 2.0 yrs; BMI= 32.4 ± 5.2 kg/m²) who were submitted to a 6-month caloric restriction. Body composition (DXA), resting EE (indirect calorimetry), physical activity EE (PAEE) and total EE (TEE) (doubly-labelled water) were measured before and after the 6 month weight loss. Blood samples were collected before and after to measure leptin and peptide YY. The results indicate that there was indeed a depression in resting EE during weight loss regardless of the type of intervention utilized. Furthermore, these findings suggest that the changes could not fully be explained by changes of FM and FFM alone and that leptin may be an important contributor to the changes of resting EE during weight loss.

Being able to effectively match energy intake to energy expenditure (EE) is an important aspect in preventing weight re-gain in the post-obese. Although it is generally agreed upon that resting EE decreases concomitantly with weight loss, there is no set standard comparing the deviations with differing weight loss protocols and additionally, controversy remains as to whether this decrease is greater than can predicted. In order to address these issues 2977 subjects were analyzed using a systematic review and the differences of both the protocol and length of various interventions in addition to sex were compared. Next, data was selected from this systematic review and 815 subjects were analyzed for weight loss-induced changes in resting EE, FM and FFM. Another subgroup of studies (n = 1450) was analyzed and compared against the Harris-Benedict prediction equation to determine whether the changes in resting EE were greater than what was expected. Finally, in order to determine which factors may be involved in regulating changes in resting EE during weight loss, a secondary analysis was performed on 28 post-menopausal women (age= 50.4 ± 2.0 yrs; BMI= 32.4 ± 5.2 kg/m²) who were submitted to a 6-month caloric restriction. Body composition (DXA), resting EE (indirect calorimetry), physical activity EE (PAEE) and total EE (TEE) (doubly-labelled water) were measured before and after the 6 month weight loss. Blood samples were collected before and after to measure leptin and peptide YY. The results indicate that there was indeed a depression in resting EE during weight loss regardless of the type of intervention utilized. Furthermore, these findings suggest that the changes could not fully be explained by changes of FM and FFM alone and that leptin may be an important contributor to the changes of resting EE during weight loss.

Being able to effectively match energy intake to energy expenditure (EE) is an important aspect in preventing weight re-gain in the post-obese. Although it is generally agreed upon that resting EE decreases concomitantly with weight loss, there is no set standard comparing the deviations with differing weight loss protocols and additionally, controversy remains as to whether this decrease is greater than can predicted. In order to address these issues 2977 subjects were analyzed using a systematic review and the differences of both the protocol and length of various interventions in addition to sex were compared. Next, data was selected from this systematic review and 815 subjects were analyzed for weight loss-induced changes in resting EE, FM and FFM. Another subgroup of studies (n = 1450) was analyzed and compared against the Harris-Benedict prediction equation to determine whether the changes in resting EE were greater than what was expected. Finally, in order to determine which factors may be involved in regulating changes in resting EE during weight loss, a secondary analysis was performed on 28 post-menopausal women (age= 50.4 ± 2.0 yrs; BMI= 32.4 ± 5.2 kg/m²) who were submitted to a 6-month caloric restriction. Body composition (DXA), resting EE (indirect calorimetry), physical activity EE (PAEE) and total EE (TEE) (doubly-labelled water) were measured before and after the 6 month weight loss. Blood samples were collected before and after to measure leptin and peptide YY. The results indicate that there was indeed a depression in resting EE during weight loss regardless of the type of intervention utilized. Furthermore, these findings suggest that the changes could not fully be explained by changes of FM and FFM alone and that leptin may be an important contributor to the changes of resting EE during weight loss.

Background. Roux-en-Y gastric bypass (RYGB) is an effective treatment for obesity, where gut peptides such as ghrelin, glucagon-like peptide-1 (GLP-1) and peptide YY (PYY) play an instrumental role in reduced appetite after RYGB. This systematic review and meta-analysis aimed to establish the magnitude of change of ghrelin, GLP-1, PYY and appetite sensation following RYGB. Methods. A systematic search was conducted in Medline Ovid, Embase, Scopus, and Cochrane Central Register of Controlled Trials up until March 2021. Two independent reviewers screened articles for studies that evaluated ghrelin, GLP-1, PYY or appetite sensation via visual analogue scales (VAS) before and after RYGB in adults. Risk of bias was assessed with the quality assessment tool for before-after studies with no control group from the National Heart, Lung and Blood Institute (NHLBI). A multilevel model with random effects for study and follow-up time points nested in study was fit to the data. The model included kilocalorie consumption as a covariate and time points as moderators. Results. Among the 2,559 articles identified, 47 met the inclusion criteria, among which k=19 evaluated ghrelin, k=40 GLP-1, k=22 PYY and k=8 appetite sensation via VAS. Our results indicate that fasting ghrelin levels are decreased 2 weeks post-RYGB (p = .005) but do not differ from baseline from 6 weeks to 1-year post-RYGB. Postprandial ghrelin levels at 6 months and 1-year post-RYGB were not different from pre-surgical values (p = .51). Fasting GLP-1 levels were not different from pre-surgical levels up to 2 years post-RYGB. Postprandial levels of GLP-1 increased significantly from 1 week (p < .001) to 2 years post-RYGB (p < .01) compared to before surgery. Compared to pre-RYGB levels, fasting PYY increased at 6 months (p = .034) and 1 year (p = .0299) post-surgery and postprandial levels were increased up to 1 year (p < .01). Heterogeneity was significant in most analyses. Insufficient data on appetite sensation was available to be meta-analyzed. Conclusion. Our analyses illustrate the magnitude of change of ghrelin, GLP-1 and PYY before and after RYGB surgery. Importantly, between study heterogeneity within the current literature warrants more standardized protocols and studies with longer follow-up periods for better comprehension of changes in gut peptides following RYGB surgery.

<p>This thesis examines the effect of surgical weight loss on gut and adipose tissue peptides involved in appetite regulation and energy homeostasis in morbidly obese humans. Roux-en-Y gastric bypass (RYGBP) is the gold standard operation used for effective long-term weight loss and improved health. The exact mechanisms for this outcome are under investigation.</p><p>We measured ghrelin, a recently discovered hunger hormone, insulin, adiponectin and leptin along with anthropometry measures in 66 morbidly obese patients prior to and 6 and 12 months after RYGBP. Impressive weight loss occurred postoperatively as did alterations in the peptides. Consistent correlations were found between weight, leptin, ghrelin and insulin. The main findings were low ghrelin concentrations in obesity and an increase after RYGBP.</p><p>We explored inflammatory proteins C-reactive protein (CRP), serum amyloid A and interleukin-6 before and during massive weight loss 6 and 12 months after RYGBP in morbidly obese subjects. The studied proteins declined after surgery and a correlation between CRP and homeostatic model of assessment for insulin resistance, independent of BMI, strongly linked insulin resistance and inflammation. CRP declined most in insulin-sensitive subjects.</p><p>We examined the excluded stomach mucosa and vagus nerve by measuring gastrin, pepsinogen I (PGI), pancreatic polypeptide (PP) and ghrelin levels during week 1 and year after RYGBP. Ghrelin levels rose with weight loss but declined 24-hours after surgery, like PP, indicating transient vagal nerve damage. Low levels of gastrin and PGI suggest a resting mucosa.</p><p>We evaluated gut peptides: peptide YY (PYY), glucaogon like peptide-1 (GLP-1), pro-neurotensin (pro-NT) and PP, in lean (young and middle-aged), obese and postoperative RYGBP subjects pre- and postprandially. RYGBP subjects had exaggerated levels of PYY and GLP-1 postprandially and higher basal proNT levels, implying a ‘satiety peptide tone’ that may contribute to the maintenance of weight loss.</p><p>In summary, RYGBP results in marked weight loss and alterations in gut and adipose tissue peptides involved in appetite regulation and energy homeostasis. These postoperative peptide changes may contribute to impressive weight loss observed after RYGBP.</p>