Sirt3ko mice

All animal studies were approved by the University of Pittsburgh Institutional Animal Care and Use Committee (IACUC) and conducted in accordance with the guidelines and regulations set forth in the Animal Welfare Act (AWA) and PHS Policy on Humane Care and Use of Laboratory Animals. Sirt3KO mice on a 129S1/SvImJ strain background were obtained from Jackson Laboratories (Bar Harbor, ME). Sirt5KO mice, also from Jackson Laboratories but on a mixed background, were backcrossed onto the 129S1/SvImJ strain. 129S1/SvImJ mice were used as wild-type controls. All experimental animals were males ages 10–14 weeks and were fasted for 16 hr prior to tissue collection (N = 5).

129 wild-type (WT) male mice of 8 weeks old were purchased from Department of Laboratory Animal Science of Peking University (Beijing, China) and SIRT3-knockout (SIRT3-KO) mice were purchased from the Jackson Laboratories (USA). SIRT3-overexpression (SIRT3-LV) mice were obtained by injection to the 129 WT mice SIRT3-overexpression-lentivirus (JIKAI GENE, Shanghai, China) via caudal vein. HKL were intraperitoneally injected in 129 WT mice for 42 days (25 mg/Kg, once a day) as the HKL treatment group. All experimental mice anesthetized with isoflurane (1%) were implanted subcutaneously corresponding osmotic pumps beforehand filled with AngII or saline for 42 days. The osmotic pumps infused with appropriate doses of AngII in sterile saline (2000 ng/kg per min) or saline according to the directions were placed in sterile 0.9% saline at 37°C for 60 hours to prime. Together, the experimental animals were randomly assigned to eight groups: WT+saline group, WT+AngII group, SIRT3-KO+saline group, SIRT3-KO+AngII group, SIRT3-LV+saline group, SIRT3-LV+AngII group, WT+HKL+saline group, WT+HKL+AngII group. Mice were sacrificed at the end of the experiment and the kidneys were used to assay for renal fibrosis. In addition, Blood pressure were measured by Tail-Cuff way (BP2010A, softron, Japan) before modeling and death.

The animal experimental protocol complied with the Animal Management Rules of the Chinese Ministry of Health (Document No. 55, 2001) and was approved by the Animal Care and Use Committee of Shandong University. The SIRT3-KO mice were purchased from the Jackson Laboratories (USA). 129 Wild-type mice at 6–7 weeks of age were purchased from the Department of Laboratory Animal Science of Peking University and served as controls (Beijing, China). All animals were maintained at the key Laboratory of Cardiovascular Remodeling and Function Research at Qilu Hospital of Shandong University.