Attenuation by Time-Restricted Feeding of High-Fat and High-Fructose Diet-Induced NASH in Mice Is Related to Per2 and Ferroptosis

Figure 7

Ferroptosis occurs in HFHFD-induced NASH and promotes the progression of NASH. (a) Fluorescence staining showing lipid peroxidation (LPO) levels in the livers of the FA+Veh and FA+Lip-1 groups. The nuclei were labeled with DAPI (left) and LPO (middle) was labeled by C11-Bodipy (original magnification, 400x; scale bar, 50 μm). (b) Liver MDA concentrations were measured with a commercial kit. (c) Acsl4 and Gpx4 protein levels were measured with western blotting, with GAPDH used as a control. (d) Liver GSH concentrations were measured by a commercial kit. (e) ALT and AST levels were determined by commercial kits to evaluate the levels of liver injury. (f, g) Serum and liver TG, TC, and FFA levels are depicted to demonstrate the lipid concentrations between the two groups. (h) Pathologic findings in liver sections. Liver paraffin sections were stained with H&E (original magnification, 200x; scale bar, 100 μm), oil red O (original magnification, 400x; scale bar, 50 μm), and Sirius red (original magnification, 100x; scale bar, 200 μm) to determine the levels of steatosis, inflammation, and fibrosis in the FA+Veh and FA+Lip-1 groups. Data are presented as . , , and ; ns: not significant (also refer to Figure S3).

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