Su1885 Insulin Resistance Is a Key Factor for Metabolic and Inflammatory Biomarkers in Children Obesity

Abstract

protein integrity. Hypothesis: Epithelial cell Hsp70 overexpression maintains tight junction protein levels in spite of high fat diet. Materials & Methods: Five week old male mice, [non-transgenic (NTG; n=19) and Hsp70 transgenic littermates (TG; n=14; villin promoter driven epithelial cell Hsp70 expression)] were randomly divided into low fat (LF; 10 kcal% fat) or high fat (HF; 60 kcal% fat) diet groups (NTG/LF, NTG/HF, TG/LF, TG/HF). All mice continued on the protocol for 12 weeks. At week 13, colonic scrapings were collected and homogenized for determining Hsp70 levels (ELISA, R&D Systems) and tight junction protein expression (Western blot). Western blots probed for occludin (Life Technologies) and constitutive heat shock protein 70 (HSC70, Santa Cruz Biotech) were run using pooled samples of each group (n= 6-10 per group). Densitometry readings were completed using a ChemiDoc MP Imager (Bio-Rad) and data normalized using HSC70 as a loading control with NTG/LF as the reference sample. Hsp70 data was analyzed by 2-way ANOVA and presented as a mean ± SEM. Results: Measurement of colonic Hsp70 levels showed a significant (p < 0.05, Fig. 1), 100 fold, increase in the TG group compared to its NTG littermates. Western blot for occludin using pooled samples representative of each group suggested TG mice expressed an increased amount of this protein compared to their NTG littermates regardless of diet (Fig. 2). Discussion: Hsp70 TG mice demonstrated increased levels of occludin, suggesting that it may enhance gut barrier function by increasing tight junction proteins independent of diet. Further examination of Hsp70's affect on colonic tight junction proteins may provide a possible target for the prevention and treatment of obesity