Neuroprotective Potential of Fasudil Mesylate in Brain Ischemia-Reperfusion Injury of Rats

Abstract

We previously reported that inhibition of Rho-kinase (ROCK) by hydroxyl fasudil improves cognitive deficit and neuronal damage in rats with chronic cerebral ischemia (Huang et al., Cell Mol Neurobiol 28:757-768, 2008). In this study, fasudil mesylate (FM) was investigated for its neuroprotective potential in rats with ischemia following middle cerebral artery occlusion (MCAO) and reperfusion. The effect of fasudil mesylate was also studied in rat brain cortical and hippocampal slices treated with oxygen-glucose deprivation (OGD) injury. Gross anatomy showed that cerebral infarct size, measured with 2,3,5-triphenyltetrazolium chloride (TTC) staining, was significantly smaller in the FM-treated than in the non-FM-treated ischemic rats. In the brain regions vulnerable to ischemia of ischemic rats, fasudil mesylate was also found to significantly restore the enzyme protein expression level of endothelial nitric oxide synthase (eNOS), which was decreased in ischemia. However, it remarkably reduced the protein synthesis of inducible nitric oxide synthase (iNOS) that was induced by ischemia and reperfusion. In rat brain slices treated with OGD injury, fasudil mesylate increased the neuronal cell viability by 40% for cortex and by 61% for hippocampus, respectively. Finally, in the presence of OGD and fasudil mesylate, superoxide dismutase (SOD) activity was increased by 50% for cortex and by 58% for hippocampus, compared to OGD only group. In conclusion, our in vivo study showed that fasudil mesylate not only decreased neurological deficit but also reduced cerebral infarct size, possibly and at least partially by augmenting eNOS protein expression and inhibiting iNOS protein expression after ischemia-reperfusion.

Neuroprotective and Antioxidative Effect of Cactus Polysaccharides In Vivo and In Vitro

“…The middle cerebral artery was occluded as described before (Longa et al 1989;Li et al 2009). Reperfusion was induced after 2 h MCAO by filament withdrawal.…”

“…Cerebral infarct size was assessed with TTC staining method (O'Donnell et al 2004;Li et al 2009). After 2 h MCAO and 24 h reperfusion, all animals were anesthetized and the brains quickly isolated and sectioned into consecutive 2-mm-thick coronal slices using a Vibratome (Campden Instruments, USA).…”

“…All brain slices of each experimental group were analyzed for the infarct size using the Image-Pro plus 5.0 analysis software. Percentage infarct size was calculated as described (Swanson et al 1990;Li et al 2009…”

“…After rabbit anti-rat iNOS polyclonal antibody (Santa Cruz, USA) (dilution 1:100) was applied to the samples and incubated for 24 h at 4°C, the sections were incubated with biotinylated goat anti-rabbit IgG (1:100) for 1 h at 37°C, followed by the steps according to SABC (HRP) kit protocol. The sections were subsequently incubated with diaminobenzidine (DAB) 0.5 g/l and observed under light microscope (Li et al 2009). …”

Fasudil, a Rho-associated protein kinase inhibitor, attenuates retinal ischemia and reperfusion injury in rats

“…Recently, evidence has accumulated suggesting that abnormal activation of ROcK plays an important role in various pathological conditions, including cerebral and coronary vasospasm, hypertension, vascular inflammation and remodeling, and arteriosclerosis (8). Studies with fasudil and the structurally unrelated inhibitor Y-27632 have indicated beneficial effects of selective ROcK inhibition in experimental models of ischemic diseases, including cerebral vasospasm, and end-organ (renal, liver, colon, brain, intestine) I/R injury (9)(10)(11)(12)(13)(14). Regarding retinal I/R injury, Hirata et al have recently reported that administration of Y-27632 attenuates neuronal cell death and dramatically inhibits leukocyte infiltration and endothelial disarrangement after transient retinal ischemia in rats (15).…”