Gm6001

Anti-IL-1β, anti-IL-6, anti-TNFα and anti-CINC-1 antibodies were supplied by R&D Systems Inc. (USA). Indomethacin, AACOCF₃ and PACOCF₃ were purchased from Biomol Research Laboratories (USA). GM6001 was supplied by USBiological (USA); whereas L-NMMA, HOE 140, Lys-(Des-Arg9,Leu8)-bradykinin, promethazine, methysergide, BQ-123, BQ-788 and fucoidan were purchased from Sigma-Aldrich Co. (USA). Celecoxib was supplied by Searle and Co (Puerto Rico). Zileuton was purchased from Abbott Laboratories (Zyflo®, USA). Carrageenin was purchased from Marine Colloids.

The broad spectrum MMP-inhibitor GM6001 (USBiological, Swampscott, Massachusetts) was used and prepared as previously described with slight modifications (Gursoy-Ozdemir et al., 2004 (link); Wang and Tsirka, 2005 (link); Chen et al., 2009 (link)). Specifically, mice were injected intraperitoneally with either 50 mg/kg GM6001 diluted in 3% DMSO and 2% cyclodextrin in saline to a total volume of 200 µl, or with 200 µl vehicle (3% DMSO and 2% cyclodextrin in saline). This type of systemic application of GM6001 was used before and shown to successfully inhibit MMPs activity in the brain tissue (Gursoy-Ozdemir et al., 2004 (link); Wang and Tsirka, 2005 (link); Chen et al., 2009 (link)). For the healthy brain group, the injections were applied daily for 7 days, starting 1 h after MD/no MD. For the stroke group, treatment started 1 h after stroke induction, with an additional injection 24 h after stroke in one group of animals.