Hydrochloride Fasudil Inhibits Inflammation And Attenuates Brain Injury In Rats After Intracerebral Hemorrhage

Objective To investigate the effect of hydrochloride fasudil(HF)on inflammation and brain injury in rats after intracerebral hemorrhage(ICH).Methods 126 Male Wistar rats were randomly divided into normal group(Normal),sham operated-group(Sham),intracerebral hemorrhage group(ICH)and hydrochloride fasudil treatment group(ICH/HF).The rat model was made by injecting non-anticoagulant autologous artery blood into the right caudate nucleus.Samples were collected on 1,3,7 and 14 day.Bederson was used to score neurological deficit in rats;Levels of Rho-associated protein kinase 2(ROCK2)mRNA and protein around the site of the hematoma were measured by quantitative real-time polymerase chain reaction(QRT-PCR)and enzyme-linked immunosorbent assay(ELISA),respectively.Interleukin(IL)-6 and tumor necrosis factor(TNF)-a in serum were detected by ELISA;The changes of brain water content were measured with dry/wet weight method;Hematoxylin eosin(HE)and nissl staining were performed to observe the inflammatory cells infiltration and the neuronal morphological changes around the hematoma.HE staining was performed to observe the morphological changes and measure the lumen area and the wall thickness of the basilar artery by using Motic Digital Medical Image software.Results The neurological deficit scores on 1,3 and 7 day in the ICH group and ICH/HF group were significantly increased as compared with those in the sham-operated group;those on 1 and 3 day in the ICH/HF group were significantly decreased as compared with that in the ICH group(P<0.05).Following ICH,levels of-ROCK2 in mRNA and protein were significantly increased from day 1 to day 7,with a peak at day 3.ROCK2 levels were significantly lower in rats treated with HF than in rats treated with saline(P<0.05).When comparing rats in the ICH group with control and sham-operated animals,IL-6 levels were significantly higher on days 1,3 and 7 and TNF-a levels and brain water content were significantly higher on days 1,3,7 and 14.Administration of HF significantly reduced levels of IL-6 and TNF-a and reduced brain water content on days 1,3 and 7 but not on day 14(P<0.05).In the early stage of ICH(1 and 3 day),mass neutrophils infiltration developed in the perihematomal areas.However,neutrophils infiltration in ICH/HF group was significantly decreased compared with ICH group on 1 day(P<0.05).After ICH,nissl staining showed neuron death and loss in the perihematomal areas at all time points;but HF treatment can significantly attenuate neuron loss(P<0.05).On day 1 and 3 in the ICH group and on day 7 in the ICH/HF group,the ratios of lumen area and blood vessel area were significantly lower than the sham-operated and normal groups(P<0.05);On day 1,3 and 7 in the ICH group and on day 7 in the ICH/HF group,the ratios of wall thickness and diameter length were significantly higher than the sham-operated and normal groups(P<0.05);After FH treatment,On day 1 and 3,the ratios of lumen area and blood vessel area significantly increased,and the ratios of wall thickness and diameter length significantly decreased compared with the ICH group on day 1 and 3(P<0.05).Conclusions HF exerts neuroprotective effects in a rat model of ICH by down-regulating expression of ROCK2,reducing neutrophil infiltration and production of inflammatory cytokines,decreasing brain edema,attenuating loss of neurons and improving neurological outcome;ICH may induce cerebral vasospasm,and HF may attenuate cerebral vasospasm following ICH.