Development of a Pediatric Model of Nafld in Neonatal Iberian Pigs

Author

Gabriella Veronica Hernandez, California Polytechnic State University, San Luis ObispoFollow
Victoria Alice Smith, California Polytechnic State University, San Luis ObispoFollow
Morgan Coffin, California Polytechnic State University, San Luis ObispoFollow
Daniel Columbus, Prairie Swine Center, Inc., Saskatoon, SKFollow
Matthew Burd, California Polytechnic State University, San Luis ObispoFollow
Kimberly Sprayberry, California Polytechnic State University, San Luis ObispoFollow
Mark Edwards, California Polytechnic State University, San Luis ObispoFollow
Daniel Peterson, California Polytechnic State University, San Luis ObispoFollow
Darin Bennett, California Polytechnic State University, San Luis ObispoFollow
Robert Fanter, California Polytechnic State University, San Luis ObispoFollow
Christopher Kitts, California Polytechnic State University, San Luis ObispoFollow
Michael La Frano, California Polytechnic State University, San Luis ObispoFollow
Margaret Rice, California Polytechnic State University, San Luis ObispoFollow
Douglas Burrin, Baylor College of MedicineFollow
Magdalena Maj, California Polytechnic State University, San Luis ObispoFollow
Rodrigo Manjarin, California Polytechnic State University, San Luis ObispoFollow

DOI: https://doi.org/10.15368/theses.2019.107
Available at: https://digitalcommons.calpoly.edu/theses/2097

Date of Award

6-2019

Degree Name

MS in Agriculture - Animal Science

Department/Program

Animal Science

College

College of Agriculture, Food, and Environmental Sciences

Advisor

Rodrigo Manjarin

Advisor Department

<--Please Select Department-->

Advisor College

College of Agriculture, Food, and Environmental Sciences

Abstract

The prevalence of non-alcoholic fatty liver disease (NAFLD) in children has increased over the past decades, creating a need for animal models that recapitulate the features of the pediatric disease. Iberian pigs have a leptin-resistant phenotype characterized by hyperleptinemia, hyperphagia, and extreme adipogenesis. We hypothesized that neonatal Iberian pigs fed a high fat high-fructose (HFF) diet will develop a pattern of liver injury resembling pediatric NAFLD. In addition, we sought to determine if a mixture of probiotics would prevent the disease. Animals were fed 1 of 4 diets containing (g/kg body weight × d) 0 g fructose, 11 g fat and 199 kcal (CON-N; n=8), 22 g fructose, 16 g fat and 300 kcal (HFF2-N; n=8), CON + probiotic (CON-P; n=6), or HFF2 + probiotic (HFF2-P; n=6) every 6 h for 70 d. The probiotic mixture (6.2 × 10⁴ cfu/mL) contained Pediococcus acidilactici, Pediococcus pentosaceus, Lactobacillus plantarum and Bacillus amyloliquefaciens. Body weight was recorded every 3 d. Serum markers of liver injury and dyslipidemia were measured on d 40 and 65 at 2 h post feeding. Fasting leptin, insulin, glucose and homeostatic model assessment (HOMA) values were assessed on d 70. Liver and skeletal muscle (longissimus dorsi) were collected on d 70 for histology, triacylglyceride (TAG) quantification, relative gene expression, and Western blot analysis. Metabolomic analysis was performed on liver tissue and plasma. Body weight was not significantly greater in HFF fed pigs compared to CON. Leptin, alanine and aspartate aminotransferases, alkaline phosphatase, lactate dehydrogenase and total bilirubin were increased (P ≤ 0.001), and high and low density lipoproteins decreased (P ≤ 0.05) in HFF2-N and HFF2-P. Livers in HFF2-P and HFF2-N had higher relative weight and TAG (P ≤ 0.001), micro and macrovesicular steatosis, ballooning degeneration, Mallory-denk bodies, inflammation and necrosis, increased gene expression of TNFα, TGFβ, IL1α and PPARγ (P ≤ 0.001), and decreased ChREBP (P ≤ 0.001). A probiotic affect was seen as pigs fed CON-P and HFF2-P had higher insulin and HOMA values were increased (P ≤ 0.05). Western blot analysis showed dysregulation of autophagy in liver of pigs fed CON-P and HFF2-P, and in skeletal muscle of pigs fed CON-N and HFF2-N. Metabolomic analysis demonstrated dysregulation of one-carbon metabolism, the tricarboxylic acid cycle (TCA), the urea cycle, and amino acid metabolism of pigs fed HFF2 diets compared to CON diets. In conclusion, Iberian pigs fed a HFF diet recapitulate many pediatric NAFLD-associated features, in the absence of obesity and independently of probiotic supplementation, suggesting a potentially suitable model for pediatric NAFLD research. Furthermore, probiotic supplementation did not ameliorate the onset of NAFLD when fed in conjunction with a HFF diet.