Study On The Mechanism Of Rho Kinase Inhibitors On Cerebral Protection In Rats With Cerebral Hemorrhage

Objective:To investigate the effect and mechanism of Rho kinase inhibitor on cerebral protection of intracerebral hemorrhage rats.Methods:One hundred and twenty healthy adult male Sprague-Dawlay(SD)rats was randomly divided into 3 groups:sham operation group,intracerebral hemorrhage group(ICH)and intervention group.The intervention group was divided into two subgroups:low-dose fasudil hydrochloride intervention group(LD)and high-dose fasudil hydrochloride intervention group(HD).In the ICH group and the intervention group,an animal model of ICH was established by stereotactic injection of non-anticoagulated autologous blood.In the sham operation group,only a microinjection needle was used to penetrate the right caudate nucleus.The sham operation group and the ICH group were intraperitoneally injected with 2 ml normal saline 30 minutes after the completion of the model,and then administered once every 24 hours.In the intervention group,Fasudil hydrochloride 10 mg/kg(LD)and 30 mg/kg(HD)were injected intraperitoneally 30 min after the completion of the model,and then administered once every 24 hours.The brains were decapitated at the corresponding time points,HE staining was used to observe the formation of hematoma and the infiltration of inflammatory cells around the hematoma.The water content of brain tissue was measured by dry-wet weight method.The levels of IL-6 and TNF-? in brain tissue were detected by ELISA.The expression of ROCK2,MYPT-1,P-MYPT-1,Bcl-2,Bax,Caspase-3 and AQP-4 protein in the perihematoma tissue was detected by Western blot.Result:1.The expression of ROCK2 protein around the hematoma at 12h,1d,and 3d after cerebral hemorrhage in rats was significantly higher than that in the sham group(P<0.05).The water content,relative content of IL-6 and TNF-? in brain tissue at 12h,Id,3d,5d,and 7d after cerebral hemorrhage were significantly higher than those in sham-operated group(P<0.05).The expression of Bcl-2,Bax,and Caspase-3 protein around the hematoma at 12h,1d,3d,5d,and 7d after intracerebral hemorrhage was significantly higher than that in the sham group(P<0.05).2.The expression of ROCK2 around the hematoma in the intervention group was significantly decreased compared with the ICH group at 12h,Id,and 3d,and the difference was statistically significant(P<0.05).The expression of ROCK2 around the hematoma in the HD 12 h after intervention was significantly lower than that in the LD at 12 h after intervention(P<0.05)There was no significant difference in the expression of ROCK2 around the hematoma Id and 3d after intervention in the intervention group.3.The level of MYPT-1 phosphorylation after ICH was significantly higher than that in the sham group and gradually decreased with time.The phosphorylation level of MYPT-1 in the intervention group was significantly lower than that in the cerebral hemorrhage group.And the effect of high-dose fasudil hydrochloride intervention group(HD)is more obvious.4.The expression of apoptosis-related protein Bcl-2 in the intervention group was significantly higher than that in the ICH group,and the expression of Bax protein was significantly reduced.AQP-4 protein expression in the intervention group was significantly reduced compared with the ICH group(12h).Conclusion:Fasudil hydrochloride can reduce the brain edema after ICH by inhibiting the expression and function of ROCK2 in the brain tissue,and inhibit the secretion of inflammatory factors and apoptosis to exert neuroprotection.