Histological analysis
Paraformaldehyde-fixed periepididymal fat and liver were
paraffin-embedded, sectioned, and stained with Mayer’s hematoxylin-eosin
(Sigma-Aldrich Co., St. Lousi, MO, USA). The NAFLD activity score and
fibrosis staging system was applied to liver sections for scoring of
steatosis, lobular inflammation, hepatocyte ballooning, and fibrosis as
outlined by Kleiner et al (Kleiner et al. , 2005). Individual
adipocyte size was calculated with ImageJ software (Schneider, Rasband
and Eliceiri, 2012). All histological assessments were performed by a
pathologist blind to treatment.