Zileuton

Thirty rats were divided into five groups (n = 6/group): Sham, SPC + SBI, Vehicle + VPC + SBI, Manoalide + VPC + SBI, and Zileuton + VPC + SBI. The PLA2 inhibitor, Manoalide (Cayman Chemical, Ann Arbor, MI) was dissolved in ethanol to obtain a stock solution with concentration of 4.8 mM. Manoalide was added to Naja sputatrix venom 1:20 and incubated at 42 °C for 40 mins to inactivate PLA2^{48 (link)–51 (link)}. The PLA2 inactivated venom was injected subcutaneously in the nape of the neck for 3 consecutive days. The selective 5-LOX inhibitor, Zileuton (Santa Cruz Biotechnology, Santa Cruz, CA) was dissolved in ethanol to obtain 20 mg/mL stock solution. Zileuton (0.5 mg/kg) was injected subcutaneously, in a different spot that was included within the same 2 × 2 cm rectangular area in the nape of the neck, 30 mins before each VPC injections^{54 (link)–57 (link)}. Brain samples were collected 24 h after SBI for western blot analysis.

Murine-recombinant leptin, rabbit anti-mouse CXCL1 antibody (catalog # 250-11), and control rabbit IgG (catalog # 500-P00) were purchased from Peprotech, Inc. (Rocky Hill, NJ, USA). Rapamycin was obtained from Sigma-Aldrich, Inc. (Saint Louis, MO, USA). Zileuton was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). U-75302 was purchased from Cayman Chemical (Ann Arbor, MI, USA). Osmium tetroxide was provided from Ted Pella, Inc. (Redding, CA, USA).