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. 2022 Oct 9;12(16):7158–7179. doi: 10.7150/thno.78376

Figure 7.

Figure 7

PA-S14 ameliorates renal fibrosis and tubular cell senescence in adriamycin nephropathy. (A) Diagram shows the experimental design. Red arrows indicate the time points for injection of adriamycin (ADR). Green line indicates PA-S14 treatment (0.5 mg/kg or 1.0 mg/kg body weight). (B) Urinary albumin levels in different groups at 3 weeks after ADR. Urinary albumin was expressed as milligrams per milligram creatinine. **P < 0.01 versus control mice; ##P < 0.01 versus ADR mice; ††P < 0.01 versus ADR mice (n = 5). (C) Quantitative analysis of tubular injury in different groups as indicated. Kidney sections were subjected to PAS staining. At least 10 randomly selected fields were evaluated under 400× magnification and results were averaged for each animal. ***P < 0.001 versus control mice; ###P < 0.001 versus ADR mice; †††P < 0.001 versus ADR mice (n = 5). (D) Graphical representations of the degree of kidney fibrotic lesions in different groups after quantitative determination of Sirius red staining intensity. **P < 0.01 versus control mice; ##P < 0.01 versus ADR mice; ††P < 0.01 versus ADR mice (n = 5). (E) Representative micrographs show tubular injury and Collagen deposition in different groups as indicated. Paraffin sections were subjected to Periodic acid-Schiff (PAS) staining and Sirius red staining, respectively. Representative micrographs showing renal expression of Fibronectin and α‐SMA in different groups. Arrows indicate positive staining. Scale bar, 50 µm. (F-I) Representative western blot (F) and quantitative data showing renal expression of Fibronectin (G), Collagen I (H), and α‐SMA (I) in different groups. Numbers (1-2) indicate each individual animal in a given group. **P < 0.01 versus control mice; #P < 0.05 versus ADR mice; ††P < 0.01 versus ADR mice (n = 5). (J-N) Representative western blot and quantitative data show renal expression of active-β-catenin, p16INK4A, Klotho and KIM-1 in different groups as indicated. Numbers (1-3) indicate each individual animal in given group. **P < 0.01, ***P < 0.001versus control mice; ##P < 0.01, ###P < 0.001 versus ADR mice (n = 5). (O) Representative staining micrographs show renal expression of active β-catenin, Klotho, KIM-1, and SA‐β‐gal activity. Frozen kidney sections were stained with an antibody against active β-catenin and assessed for SA‐β‐gal activity. Paraffin kidney sections were immunostained with the antibody against Klotho or KIM-1. Arrows indicate positive staining. Scale bar, 50 µm.