Recombinant human lif

All animals were treated prophylactically with ketoprofen (10 mg/kg s.c.) and atropine (0.25 mg/kg s.c.) with two additional doses of ketoprofen at 24 and 48 h post-MCAO. Recombinant human LIF (ProSpec, Ness Ziona, Israel) (125 μg/kg) or PBS (pH 7.4) was administered intravenously at 6, 24, and 48 h post-MCAO as previously described [34 (link), 35 (link)]. Animals were randomly assigned to treatment groups, and all lab personnel administering drugs were blinded to treatments.

Induction of FCI was achieved using the permanent middle cerebral artery occlusion (MCAO) model as previously described [44 (link)]. Briefly, an incision was made near the sternum and blunt dissection was used to expose the common carotid artery. After ligating and cutting the external carotid artery, a 40-mm monofilament was introduced and fed through the internal carotid artery. The monofilament was advanced to the base of the middle cerebral artery. The monofilament was secured and the wound was closed using polypropylene sutures. Laser Doppler (Moore Lab Instruments, Farmington, CT) was used to confirm the reduction in cerebral blood flow. Animals experiencing less than a 60 % reduction in CBF were excluded from analysis. For the sham MCAO procedure, the common carotid artery was exposed without subsequent occlusion of the middle cerebral artery. Animals were administered recombinant human LIF (ProSpec, Ness Ziona, Israel) (125 μg/kg) intravenously at 6, 24, and 48 h post-MCAO. Phosphate-buffered saline (PBS) pH 7.4 was used as a vehicle control for the LIF. Rats were euthanized 24, 48, and 72 h post-MCAO for immunohistochemistry or biochemical analysis. The number of subjects in each treatment group was as follows 24 h PBS: n =3; 24 h LIF: n = 5; 48 h PBS: n = 4; 48 h LIF: n = 4; 72 h PBS: n = 12; and 72 h LIF: n = 10.