FIGURE 2.

Role of autophagy in the protective effect of sirtuin (SIRT)3 against sepsis-induced acute kidney injury. Wild-type (WT) and SIRT3^−/− mice were injected intraperitoneally (i.p.) with either 3-methyladenine (30 mg/kg, CLP + 3-MA) or C in equivalent volumes at 12.00 and 1 h before cecal ligation and puncture (CLP). Sham-operated animals served as negative controls (Sham). Kidney tissues and blood samples were collected at 24 h after CLP using the following analysis: (A) Blood urea nitrogen (BUN) and (B) serum creatinine (SCr) were analyzed using commercial kits, (C) western blotting of LC3-I/II, Beclin-1 (BECN1) and cleaved caspase-3 in the whole kidney, (D–F) relative protein levels were determined after normalization to β-actin. (G) The collected kidneys of mice were stained with hematoxylin and eosin. Scale bars: 50 μm. (H) Quantitative evaluation of morphological tubular damage. (I,J) Apoptosis of renal tubular cells of mice was measured by the terminal UTP nick end labeling (TUNEL) assay and quantified. Scale bars: 50 μm. All data were expressed as the mean ± SEM (n = 8 mice/group). ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001 versus Sham in WT or SIRT3^−/− mice. ^#P < 0.05, ^##P < 0.01.