Dppiv inhibitor cocktail

Rats were weighed and randomly assigned to either chow (n=30) or HFD (n=61) treatment groups (t = 0 weeks) and maintained on these diets for 5 weeks. At t = 5 weeks, rats were weighed, and fat mass measured via an EchoMRI™ Body Composition Analyzer (Echo Medical Systems). HFD rats were randomly assigned to either receive OFS in their drinking water (n=31) or not (HFD; n=30). Rats were maintained on these diets for the remaining 3 weeks (Fig. 1a). At t = 8 weeks, overnight fasting body weight, fat mass, and blood were collected two days before terminal study. Adiposity was calculated as a proportion of fat mass to total body weight. Rats were randomly assigned for terminal postprandial tissue collections at timepoints 0, 2, 4, 6, and 8h post refeeding (n=4–6 per group, per timepoint). For the refeeding experiment, rats were overnight fasted, and the following morning given an isocaloric meal of their respective diet, then sacrificed according to their timepoint post refeeding. Rats were deeply anesthetized via isoflurane inhalation. Tissue collections included liver, visceral adipose, as well as ileum and colon mucosal scrapings and luminal intestinal contents from the ileum, cecum, and colon, that were quickly snap frozen and stored at −80°C. Plasma of portal vein and vena cava blood was collected with a DPPIV inhibitor cocktail (Millipore Sigma) and stored at −20°C.

At the conclusion of the study, rats were deeply anesthetized and portal blood, small intestinal contents and ileal tissue, colon contents, and liver collected. Approximately 1 mL of portal vein blood was placed in a 1.7 mL Eppendorf tube containing a DPPIV inhibitor cocktail (Millipore Sigma, Burlington, MA). Whole blood was centrifuged, plasma collected, aliquoted, and stored at −20 degrees C for future analysis. The small intestine and colon were located, isolated, and contents snap frozen in liquid nitrogen and stored at −80 degrees C for future analysis. Liver, small intestine, and colon tissue samples were collected, snap frozen, and stored at −80 degrees C for future analyses.