Figure 6.

HNE induces lysosomal disintegrity by activating μ-calpain in hepatocytes of Japanese macaque monkeys.A, Macroscopic findings of livers in the control (Cont) and in monkeys treated with HNE (HNE). Black arrows show regional discoloration. B, Hematoxylin and eosin staining and HNE immunostaining of liver tissue from the control group (Cont) and HNE-treated group (HNE). HNE immunoreactivity was observed in hepatocytes. C, The expression of liver HNE protein adducts in the control (Cont) and in monkeys treated with HNE (HNE) was evaluated by a Western blotting analysis. In the Western blotting analysis, HNE is depicted as HNE protein adducts of various molecular weights. P, protein marker. D, Bands of panel C are quantified and shown as relative fold ratios. E, Alterations in ALT levels before the HNE treatment and increases after the HNE treatment are shown. F, Comparison of immunofluorescence staining in the control group (Cont) and HNE-treated group (HNE). Blue, DAPI; green, CTSB; red, LAMP2; yellow, merge. The yellow square shows a magnified image of the liver in the control group, whereas the red square shows a magnified image of the liver in the HNE-treated group. G, The area of particles stained with LAMP2 is shown in 20× image for each monkey. H, The number of stained areas that were 10 μm² or larger in panel G is shown. I, Electron microscopy images of livers in the control (Cont) and HNE-treated groups (HNE). Black squares show magnified images of membrane-bound lysosomes in the control (Cont) and disruption of the lysosomal limiting membrane in the HNE-treated group (HNE). L, lysosome. J, A Western blotting analysis showing the expression of activated μ-calpain in the livers of the control (Cont) and HNE-treated groups (HNE). P, protein marker. K, Bands of panel J are quantified and shown as relative fold ratios.