Necroptosis was recently discovered as one form of programmed cell death (PCD) and could be specifically inhibited by
necrostatin-1. The aim of this study was to examine the effect of
necrostatin-1 on
brain injury and investigate the role of
necrostatin-1 on the other two types PCD (apoptosis and autophagic cell death) in a mouse
intracerebral hemorrhage (ICH) model. Male ICR mice received an infusion of type IV
collagenase to induce ICH or saline as control into the left striatum. In the presence of vehicle, 3-MA, zVAD, and
necrostatin-1 were pretreated with a single intracerebroventricular (i.c.v.) injection in the ipsilateral ventricle 15 min before ICH, respectively. Compared with vehicle groups,
necrostatin-1 treatment significantly reduced injury volume and
propidium iodide-positive cells at 24 and 72 h after ICH. Immunoblotting analysis showed that
necrostatin-1 treatment suppressed autophagic-associated
proteins (LC3-II, Beclin-1) and maintained p62 at normal level at 24 and 72 h after ICH. In addition,
necrostatin-1 treatment enhanced the
protein level of Bcl-2 and decreased the
protein level of cleaved
caspase-3 and the
Beclin-1/Bcl-2 ratio at 24 and 72 h after ICH. Moreover, both 3-MA and
necrostatin-1 treatment could suppress cleaved
caspase-3 and LC3-II production, whereas zVAD treatment could inhibit
caspase-3 cleavage but increased LC3-II
protein levels at 72 h after ICH. Taken together, the data demonstrated for the first time that the specific inhibitor
necrostatin-1 suppressed apoptosis and autophagy to exert these
neuroprotective effects after ICH and that there existed a cross-talk among necroptosis, apoptosis, and autophagy after ICH.