RESUMEN
BACKGROUND: Nutrients affect small intestinal protein mass and metabolism, but studies on the effect of nutrients on small intestinal protein degradation are very limited due to a lack of a proper method. The objectives of this study were to establish a method to directly estimate protein degradation in isolated enterocytes from rats and to test the effect of energy substrates on protein degradation. METHODS: Male Sprague-Dawley rats (150-200 g, n>or=8 per treatment) were used. Cell viability, tyrosine release as an indicator of protein degradation, and the effect of osmolarity, 50 mmol/L glucose, 20 mmol/L beta-hydroxybutyrate, 4.7 mmol/L butyrate, and 30 mmol/L glutamine on protein degradation were measured. RESULTS: The average viability of enterocytes at time 30 minutes was 85.8% (range, 81%-94%). Tyrosine release was linear over the course of experiments, indicating constant protein degradation (R2=0.9943; p<.05). Osmolarity, glucose, and glutamine had no effect on protein degradation in isolated enterocytes. Beta-hydroxybutyrate significantly decreased it (-16%; p<.05), whereas butyrate slightly increased it (+5%; p<.05). CONCLUSIONS: A high viability and constant protein degradation indicate a successful establishment of a method to estimate protein degradation in isolated small intestinal enterocytes from rats. The large effect of beta-hydroxybutyrate suggests a potential positive role for ketone bodies to limit the loss of small intestinal protein mass by decreasing protein degradation.
