Enteroendocrine Hormonal Response After the Ingestion of Cola Beverages With Sucrose and Non-nutritive Sweeteners

Enteroendocrine Hormonal Response After the Ingestion of Cola Beverages With Sucrose and Non-nutritive Sweeteners

Enteroendocrine, Pancreatic, and Adipocyte Hormonal Response After the Ingestion of Cola Beverages With Sucrose and Non-nutritive Sweeteners in Healthy Adults. Crossover Randomized Controlled Trial

Introduction: The consumption of non-nutritive sweeteners (NNS) has been increasing in recent years, as an alternative to replace sugars and reduce the additional intake of carbohydrates, with the idea of reducing the risk of developing obesity, metabolic syndrome, and diabetes. However, recent evidence shows that their chronic intake induces endocrine alterations that may have an important contribution to the increase in body weight. Few studies have explored the acute effects of NNS beverage consumption on endocrine response, and to date, the evidence has been inconsistent regarding post-drinking effects and potential health risks. Objective: To evaluate the effect of 3 different types of cola beverages, compared with carbonated water, on glucose, insulin, glucagon, and appetite-regulating hormones during the first 120 minutes after ingestion. Methods: A triple-blind, randomized crossover controlled trial was carried out in which 20 healthy adult individuals (10 men and 10 women) were included. With a washout period of one week (7 days) and fasting for 8 hours, each participant consumed orally 355 mL of carbonated water (CAR), and the 3 different cola beverages sweetened with sucrose (SUC), aspartame/acesulfame K (ASP), and sucrose/stevia (STE), in its commercial presentation. The serum levels of glucose, insulin, glucagon, GLP-1, GIP, PYY, leptin, pancreatic polypeptide, and ghrelin were determined during the administration of each one of the drinks before the intake of the drink and later at 30, 60, 90, and 120 minutes. Statistical analysis: A descriptive analysis of the variables was performed. The global response of glucose, insulin and appetite-regulating hormones was estimated and the Area Under the Curve (AUC) was obtained using a trapezoidal model and analyzed for each outcome by one-factor ANOVA. An ANOVA for repeated measures was performed considering treatment and time as factors, and comparisons were made with the carbonated water as a control using the Bonferroni test. P values less than 0.05 were considered statistically significant. Ethical considerations: Our institution's Research, Bioethics, and Biosafety committees authorized the project. All the participants were informed about the objective, the procedures, and the possible adverse effects considered within the study, and they signed the informed consent before the start of the interventions.

Triple masking was applied, as described below. Participants: The drinks were offered in standardized disposable cups so that the participants could not visually identify the test drink. All beverages were offered at approximately 4°C and participants were asked to consume them within 10 minutes. Investigator: All participants received the mineral water (control) in the first study session. Allocation to the remaining 3 interventions was randomized for each participant, and members of the research team who administered the beverages, performed blood sampling, processed samples in the laboratory, or followed participants during the study sessions were unaware of the allocation, that is, the type of drink that was administered to each participant. Outcomes Assessor: The investigator who performed the statistical analysis did not have access to the identity of the participants or the allocation of interventions.

355ml of an orally ingested carbonated cola drink, sweetened with 142mg of aspartame and acesulfame k, combined (soft drink in its commercial presentation)

355ml of an orally ingested carbonated cola drink, sweetened with 16g of sucrose and 15.62mg of stevia (soft drink in its commercial presentation)

Before starting the study: Subjects were asked to avoid non-nutritive sweetener consumption for 48 hours prior to the study sessions. A standardized menu of 346 Kcal was prescribed for dinner and fasting for at least 8 hours prior to each study session. A 7-day washout period was left between each study session. Session 1: Weight, height, and waist circumference were measured, the BMI was calculated, and sociodemographic data of the participants and information on the habitual consumption of soft drinks were collected. In each session: The beverages were offered at approximately 4°C, in standardized disposable cups, and the participants were asked to drink them in a maximum time of 10 minutes. Heart rate, respiratory rate, and blood pressure were measured before the intervention and every 30 minutes during the observation period. The subjects who manifested adverse sensations were referred to receive medical attention and withdrawn from the session.

Glucose, insulin, glucagon, glucagon-like peptide-1 (GLP-1), peptide tyrosine tyrosine (PYY), glucose-dependent insulinotropic polypeptide (GIP), pancreatic polypeptide (PP), leptin, and ghrelin

4-5mL of venous blood was obtained from each participant and immediately transferred to the laboratory for analysis. The glucose oxidase technique was implemented to determine serum glucose levels. This technique has high intra- and inter-assay precision (CV: 2.1 and 1.73%, respectively), and high correlation with Gernon's reagent (r=0.993). For the determination of insulin, glucagon, GLP-1, PYY, GIP, PP, leptin, and ghrelin levels, implemented the Milliplex Map® immunological assay, which has an intra-assay coefficient of variation of less than 10% and an inter-assay coefficient of less than 15%, with an accuracy greater than 97%. The tests were carried out in duplicate.

Glucose, insulin, glucagon, glucagon-like peptide-1 (GLP-1), peptide tyrosine tyrosine (PYY), glucose-dependent insulinotropic polypeptide (GIP), pancreatic polypeptide (PP), leptin, and ghrelin

4-5mL of venous blood was obtained from each participant and immediately transferred to the laboratory for analysis. The glucose oxidase technique was implemented to determine serum glucose levels. This technique has high intra- and inter-assay precision (CV: 2.1 and 1.73%, respectively), and high correlation with Gernon's reagent (r=0.993). For the determination of insulin, glucagon, GLP-1, PYY, GIP, PP, leptin, and ghrelin levels, implemented the Milliplex Map® immunological assay, which has an intra-assay coefficient of variation of less than 10% and an inter-assay coefficient of less than 15%, with an accuracy greater than 97%. The tests were carried out in duplicate.

Glucose, insulin, glucagon, glucagon-like peptide-1 (GLP-1), peptide tyrosine tyrosine (PYY), glucose-dependent insulinotropic polypeptide (GIP), pancreatic polypeptide (PP), leptin, and ghrelin

4-5mL of venous blood was obtained from each participant and immediately transferred to the laboratory for analysis. The glucose oxidase technique was implemented to determine serum glucose levels. This technique has high intra- and inter-assay precision (CV: 2.1 and 1.73%, respectively), and high correlation with Gernon's reagent (r=0.993). For the determination of insulin, glucagon, GLP-1, PYY, GIP, PP, leptin, and ghrelin levels, implemented the Milliplex Map® immunological assay, which has an intra-assay coefficient of variation of less than 10% and an inter-assay coefficient of less than 15%, with an accuracy greater than 97%. The tests were carried out in duplicate.

Glucose, insulin, glucagon, glucagon-like peptide-1 (GLP-1), peptide tyrosine tyrosine (PYY), glucose-dependent insulinotropic polypeptide (GIP), pancreatic polypeptide (PP), leptin, and ghrelin

4-5mL of venous blood was obtained from each participant and immediately transferred to the laboratory for analysis. The glucose oxidase technique was implemented to determine serum glucose levels. This technique has high intra- and inter-assay precision (CV: 2.1 and 1.73%, respectively), and high correlation with Gernon's reagent (r=0.993). For the determination of insulin, glucagon, GLP-1, PYY, GIP, PP, leptin, and ghrelin levels, implemented the Milliplex Map® immunological assay, which has an intra-assay coefficient of variation of less than 10% and an inter-assay coefficient of less than 15%, with an accuracy greater than 97%. The tests were carried out in duplicate.

Glucose, insulin, glucagon, glucagon-like peptide-1 (GLP-1), peptide tyrosine tyrosine (PYY), glucose-dependent insulinotropic polypeptide (GIP), pancreatic polypeptide (PP), leptin, and ghrelin

4-5mL of venous blood was obtained from each participant and immediately transferred to the laboratory for analysis. The glucose oxidase technique was implemented to determine serum glucose levels. This technique has high intra- and inter-assay precision (CV: 2.1 and 1.73%, respectively), and high correlation with Gernon's reagent (r=0.993). For the determination of insulin, glucagon, GLP-1, PYY, GIP, PP, leptin, and ghrelin levels, implemented the Milliplex Map® immunological assay, which has an intra-assay coefficient of variation of less than 10% and an inter-assay coefficient of less than 15%, with an accuracy greater than 97%. The tests were carried out in duplicate.

Inclusion Criteria: Voluntary participation expressed through the signing of informed consent Exclusion Criteria: Pregnancy (date of last menstruation >28 days) History of diabetes mellitus or glucose intolerance, endocrinopathies, or pancreatic diseases Treatment with medications or supplements that modify glucose and insulin (eg antihypertensives, corticosteroids, hypoglycemic agents, hormonal agents, etc.) Gastrointestinal diseases or conditions that alter gastric emptying and intestinal transit Hypersensitivity to the compounds that will be used in the study