FIGURE 4.

Effect of autophagy inhibition on the protective effect of sirtuin (SIRT)3 against renal function and apoptosis mediated by the AMP-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) pathway. The C57BL/6 mice were injected intravenously (i.v.) via the tail vein with 100 μl pGC-FU vector (vector) or vector expressing SIRT3 (pSIRT3) lentiviral plasmids at 2 weeks and 24 h before cecal ligation and puncture (CLP) or sham surgery. The pSIRT3 injected mice were treated i.p. with 3-methyladenine (3-MA; 30 mg/kg, pSIRT3+3-MA) or C in equivalent volumes at 12.00 and 1 h before CLP. Kidney tissues and blood samples were collected at 24 h after CLP. (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), cleaved caspase-3, phospho (p)-AMPK, and p-mTOR in the whole kidney. (D–H) Relative protein levels were determined after normalization to β-actin, AMPK, or mTOR. All data were expressed as the mean ± SEM (n = 6 mice/group). ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001 versus Sham in each group. ^#P < 0.05, ^##P < 0.01.