Angiopoietin-like-3 knockout protects against glomerulosclerosis in murine adriamycin-induced nephropathy by attenuating podocyte loss

Figures

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  Fig. 1. Angptl3 knockout attenuates proteinuria and hypoproteinemia, and improves renal function and survival. a Time courses of urine albumin: creatinine, ratio, serum albumin level, blood urea nitrogen level, serum creatinine level and survival percentage b of Angptl3+/+ and Angptl3−/− mice with or without ADR injection (each n = 6)

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  Fig. 2. Angptl3 knockout protects renal structure. Representative PAS-stained kidney sections harvested at different time points from Angptl3+/+ and Angptl3−/− mice treated with or without ADR injection (400 ×; scale bars = 15 μm; arrowheads in the image indicate podocyte vacuolization; arrows indicate balloon adhesions; * indicates glomerulosclerosis formation)

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  Fig. 3. Angptl3 knockout protects the ultrastructure of podocytes. Representative kidney ultrastructure observed with a TEM (a) and the quantification of foot process width (b) and number (c) of Angptl3+/+ and Angptl3−/− mice treated with or without ADR injection (10,000 ×; scale bars = 1 μm; red frames indicate podocyte foot process effacement; arrows indicate podocyte microvillus transformation; arrowheads indicate foot process detachment). Data are shown as the mean ± SEM; * P < 0.05 and ** P < 0.01

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  Fig. 4. Angptl3 knockout reduces cell loss in renal glomeruli of mice with glomerulosclerosis. a Detached cells in Bowman’s capsule of Angptl3+/+ and Angptl3−/− mice treated with ADR injection for 12 weeks. The detached cells in Bowman’s capsule were analyzed under a light microscope and TEM. (PAS staining image with 400 × magnification, scale bars = 5 μm, 30 glomeruli from each mouse, 6 mice per group; transmission electron microscopy image with × 25,000 magnification, scale bars = 1 μm, 1 glomeruli from each mouse, 6 mice per group). b Apoptotic cells in glomeruli of Angptl3+/+ and Angptl3−/− mice treated with ADR injection for 12 weeks (630 ×, scale bars = 5 μm, 30 glomeruli from each mouse, 6 mice per group). Data are shown as the mean ± SEM; * P < 0.05 and ** P < 0.01

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  Fig. 5. Angptl3 knockout alleviates the loss of podocytes from glomeruli to the urine in the ADR nephropathy mouse model. a Podocyte density in glomeruli of Angptl3+/+ and Angptl3−/− mice treated with or without ADR injection for 12 weeks detected by immunofluorescence staining for the podocyte-specific markers nephrin and podocin and confocal microscopy (630 ×, scale bars = 5 μm, n = 9). The mRNA levels of the podocyte-specific markers nephrin (b) and podocin (c) in the urine of Angptl3+/+ and Angptl3−/− mice treated with ADR injection from week 1 to week 12. Data are shown as the mean ± SEM; n = 6 per group; * P < 0.05

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  Fig. 6. Angptl3 knockout attenuates the ADR-induced primary podocyte loss including detachment and apoptosis, in vitro. Primary podocytes of Angptl3+/+ and Angptl3−/− mice treated with or without ADR attached to the bottom of culture dishes (100 ×, scale bars = 30 μm, n = 6), and the rates of cell attachment in the four groups were calculated (a). Podocyte apoptosis was analyzed by flow cytometry analysis (b) and a TUNEL assay (200 ×, scale bars = 15 μm, n = 6) (c). Data are shown as the mean ± SEM; n = 6 per group; **P < 0.01 and ***P < 0.001; ADR (−): podocytes with PBS treatment; ADR (+): podocytes with ADR treatment. ADR (+): podocytes with ADR treatment