TPPU and EC5026 treatment

The sEH inhibitors TPPU was synthesized as described previously (31), and EC5026 was synthesized as described (international patent publication no. WO 2015/148954 A1). TPPU or EC5026 was dissolved either in dimethyl sulfoxide for in vitro and ex vivo treatment (TPPU only) or in 1% polyethylene glycol, molecular weight 400 (PEG-400, Thermo Fisher Scientific) for in vivo treatment. The vehicle mice received oral gavage treatment of 1% PEG-400. Mice were perfused with saline before collecting brain for biochemical analysis. In the acute TPPU or EC5026 treatment regimen, 10- to 12-week-old C57BL/6 mice received first dose of TPPU (3 mg/kg) via oral gavage 24 hours before cotreatment of LPS (3 mg/kg, i.p.) and TPPU or EC5026 (second dose). Eighteen hours after cotreatment, mice were euthanized for analysis.

For long-term TPPU treatment, 5xFAD transgenic mice received TPPU (3 mg/kg) in drinking water for either 2.5 or 4.5 months starting at 2 months of age. The vehicle mice received 1% PEG-400 in drinking water. Every 2 weeks, mice were supplied with a new water bottle containing fresh TPPU or vehicle solution.