Mice were injected intraperitoneally with vehicle (0.9% sterile saline) or AVE0991 (a nonpeptide Ang-(1–7) analogue, 10 mg/kg, MedChemExpress LLC, Monmouth Junction, NJ, USA) once a day for 30 consecutive days (see Supplementary Figure S1). The dose and route of AVE0991 administration were chosen based on previous studies.9 (link),19 (link) According to our previous findings, AVE0991 could steadily cross the blood–brain barrier via intraperitoneal injection route.9 (link),19 (link) During the whole experiment, we monitored the general health of mice and did not observe obvious adverse effects or significant changes in their body weight or food intakes. Additionally, systolic blood pressure (SBP) was measured at the beginning and the end of the treatment period using a tail-cuff method as described.19 (link) AVE0991 injection (10 mg/kg/day) did not significantly affect SBP of APP/PS1 mice at the end of the treatment period (106.27±19.09 mmHg vs 102.44±11.67 mmHg, P>0.05).
For cell experiment, AVE0991 was diluted in DMEM/F12 (Gibco, Armonk, NY, USA) to achieve a final concentration at 10 μM. Oligomeric Aβ1–42 (Abcam, Inc., MA, USA) was prepared as described,18 (link),20 (link) and diluted with DMEM/F12 to a final concentration of 5 μM. The dose of AVE0991 and Aβ1-42 in cell experiments was chosen based on previous studies.9 (link),18 (link)