Fig. 3: Rapamycin enhances mitophagy and protects against cisplatin-induced kidney injury in mice.

C57BL/6 mice (male, 8–10 weeks old) were divided into three groups for following treatments, respectively: (1) saline control; (2) cisplatin treatment for 72 h; and (3) cisplatin + rapamycin treatment for 72 h. Cisplatin was injected at 30 mg/kg. For rapamycin, 1 mg/kg rapamycin was injected 1 h prior to and 1 day after cisplatin injection. a, b Blood samples were collected for measurements of BUN and serum creatinine. c Representative histology shown by H&E staining. Tubular injury was indicated by tubular dilation/flattening, loss of brush border, sloughing of cells into tubular lumen, formation of tubular casts, tubular degeneration, and vacuolization. Scale bar, 50 μM. d Pathological score of tubular damage in cisplatin and cisplatin + rapamycin groups. Tissue injury was scored by the percentage of damaged renal tubules (0, no damage; 1, <25%; 2, 25–50%; 3, 50–75%; and 4, >75%). e Representative images of TUNEL staining. Scale bar, 50 μM. f Quantification of TUNEL-positive cells in cisplatin and cisplatin + rapamycin groups. g, i, j Renal cortex tissues were analyzed for immunoblot analysis of TOM20, TIM23, LC3II/I, p62, HSP60, PINK1, Parkin, and GAPDH. h Densitometry analysis of proteins signals on immunoblots. The protein signals were divided by GAPDH signal of the same samples to determine the ratios. Data in a, b and d, f, h, j are expressed as mean ± SD. n = 3, ^*P < 0.001, ^**P < 0.01, ^***P < 0.05 vs. the control group; ^#P < 0.05 vs. the cisplatin group