Histological Assessment of Liver Pathology

The mice liver tissues were fixed in Mildform (Fujifilm Wako Chemicals, Osaka, Japan), processed for paraffin embedding, and sectioned into 3 µm-thick sections. Each section was deparaffinized, rehydrated, and processed for hematoxylin–eosin and Sirius red-Fast Green staining. Sections were incubated in 0.04% Fast Green (Fujifilm Wako Chemicals), saturated in picric acid for 5 min, washed with distilled water, and incubated in 0.1% Sirius Red solution (Fujifilm Wako Chemicals) for 10 min. The histological features after each staining were determined using a Leica DMi8 microscope and LAS X version 3.3 (Leica Microsystems, Inc., Wetzlar, Germany). The steatosis, inflammation, and fibrosis degrees were assessed by an experienced pathologist blinded to the experimental procedures according to previously reported criteria^{33 (link),34 (link)}.