To Compare the Effects of Non-nutritive Sweeteners Intake in Subjects With T2DM

Protocol The experimental design was developed 3 times. Each intervention was performed separately for a minimum period of 7 days out and a maximum of 14 days, depending on the availability of the experimental subjects. Cross treatment design (crossover) each subject serves as its own control The first intervention corresponded to the control group, in order to maintain the ignorance of the type of sweetener they were receiving, between day 7 or maximum 14 days post-intervention the order of the test of the NNS was reversed. Therefore, during the second or third intervention they were given to drink water with sucralose or stevia respectively. For each intervention a check list was followed.

Subjects were asked to maintain their normal diet between study days and abstain from strenuous exercise and alcohol intake for 24 hours before each evaluation. The subjects attended with an 8-hour fast, without having ingested metformin or other medication, to the sampling room of the Department of Nutrition (Faculty of Medicine, University of Chile). The study was only begun if they had a capillary glycemia <140%, measured through a glucometer.

Then, they were asked to answer the analogue visual scale (VAS) questionnaire and an intravenous catheter was installed in the antecubital vein to take the blood samples at "time -10". Immediately, they were given to drink 60 ml of water or 60 ml of water with 48 mg of sucralose or an equivalent volume with 96 mg of stevia (steviol glycosides), which they had to consume in less than 2 minutes, and then answer to a question about the "sensation" that came with drinking this preparation, to make an estimate of your predisposition to the taste of the sweetener: sweet, bitter, metallic or other flavor.

After 10 minutes, at "time 0" immediately after the second blood sample, the individuals consumed a mixed test meal consisting of a cup of tea with three teaspoons of sugar, plus 126 grams of a homemade bread. , without fat, using the bread maker, brand; program 3, traditional bread.

The following blood samples were taken at times 30, 60, 90, 120, 150 and 180 minutes after "time 0". In total, 42 ml of blood was extracted, in each time the glycemia and insulin were determined (4 ml was taken at a time). Furthermore, only at time -10, 0, 30, 90 and 180 minutes, the value of GLP-1 and ghrelin was determined (at this time, 2 ml extra was extracted at a time).

In addition, VAS questionnaires were applied every 30 minutes, from "time -10" to 180 minutes for appetite determinations.

After completing the extraction of blood samples at 180 minutes and removing the intravenous catheter, the subjects were offered a meal for 30 minutes, which consisted of a variety of foods of high acceptability, previously weighed and fractionated.

Statistic analysis The normal distribution of the variables was determined by the Shapiro-Wilk test. The natural logarithm was applied to the parameters without normal distribution. The variables were expressed as mean ± standard deviation or median plus interquartile interval (Q1-Q3). The statistical significance between the differences in plasma glucose concentrations, area under the curve (AUC) of glycaemia, energy intake and macronutrients by treatment (preload water, sucralose and stevia), was evaluated through Anova of repeated samples . To analyze the differences between plasma concentrations and AUC of insulin and GLP-1 per treatment (preload water, sucralose and stevia), Friedman's test was used, followed by Wilcoxon and adjusted by Bonferroni.

To identify differences between the perception of sweet, bitter, and metallic taste by treatment (preload water, sucralose and stevia) Friedman test was used, followed by Wilcoxon and adjusted by Bonferroni.

To identify the relationships between glycemic responses and intake of food at will (energy intake and macronutrients by treatment, preload water, sucralose and stevia), the Pearson correlation factor was used, in contrast to insulinemic response, incretin GLP-1 and the visual analogue scale (VAS) and food intake at will per treatment Spearman's correlation factor was used.

The statistical analysis was performed with the SPSS 20.0 computer program (SPSS Inc., Chicago Illinois). A p <0.05 was considered statistically significant for all the analyzes.