The Effect And Mechanisms Of Escin On Neurological Function And Blood Brain Barrier Impairment In Intracerebral Hemorrhagic Mice

Objective: To evaluate the effect and mechanisms of sodium aescinate(Escin)on neurological function and blood brain barrier(BBB)impairment in intracerebral hemorrhagic(ICH)mice.Methods: 1.Male CD-1 mice were randomly divided into Control group,ICH group.The ICH model was prepared by intrastriatal injection of bacterial collagenase type VII with a stereotaxic instrument.Then the mice were administered with escin(0.45 mg/kg)by the tail vein.The concentration of escin in serum and brain tissue were detected at 0.5 h,2 h,6 h and 12 h after escin treatment.2.Male CD-1 mice were randomly divided into Control group,ICH group,Escin(0.45 mg/kg)group,Escin(0.9 mg/kg)group,Escin(1.8 mg/kg)group.The ICH model was prepared as described above.At 24 h and 72 h of ICH,Garcia test was used to evaluate the neurological function of the mice.Brain water content was observed via a wet/dry weight method.Evans blue extravasation in cerebrums were also assayed.The serum IL-1? level was detected with ELISA kit.3.Male CD-1 mice were randomly divided into Control group,ICH group,Escin(0.9 mg/kg)group,Escin(0.9 mg/kg)+IL-1?(10 ng/m L)group.The ICH model was prepared as previous method.At 24 h of ICH,Garcia test was used to evaluate the neurological function of the mice.Brain water content was evaluated via a wet/dry weight method.Evans blue extravasation in cerebrums were also assayed.The serum IL-1? level was detected by ELISA kit.And the expression of Rho A,ROCK1,I?B?,nuclear NF-?B,cytosolic NF-?B,Occludin and Claudin-5 in mouse brain tissue were investigated with Western blot.Results: 1.Escin was detected in the serum of Control group and ICH group after 0.5 h of escin administration.At 12 h after escin administration,there is still escin in serum.Escin was not detected in brain tissues at any time points in both Control group and ICH group.2.At 24 h of ICH,mice in the ICH group showed worse performances in Garcia test(P < 0.01),however,the Garcia test scores in Escin groups were significantly increased(P < 0.05 or P < 0.01).The brain water contents of the right basal ganglia in the ICH group were augmented(P < 0.01),and they were significantly recuced by escin(P < 0.01).Evans blue extravasation in the right hemisphere was enhanced in ICH mice(P < 0.01),and it was reduced by escin treatment(P < 0.05 or P < 0.01).Compared with the Control group,the levels of IL-1? in the ICH group was significantly increased(P < 0.01).Compared with the ICH group,the levels of IL-1? in Escin groups were reduced(P < 0.05 or P < 0.01).At 72 h of ICH,mice in the ICH group showed worse performances in Garcia test(P < 0.01).However,the Garcia test scores in Escin groups were significantly increased(P < 0.01).The brain water contents of the right basal ganglia in the ICH group were augmented(P < 0.05),and they were significantly decreased by escin(P < 0.05).Evans blue extravasation in the right hemisphere was enhanced in ICH mice(P < 0.01),and it was reduced in the Escin groups compared with the ICH group(P < 0.01).Compared with the Control group,the levels of IL-1? in the ICH group were significantly increased(P < 0.01).Compared with the ICH group,the levels of IL-1? in Escin group were lower than that of the ICH group(P < 0.05).3.At 24 h of ICH,mice in the ICH group showed worse performances in Garcia test(P < 0.01).However,the Garcia test scores in Escin groups were significantly increased(P < 0.01).IL-1? recombinant protein reversed the effect of escin on Garcia test scores(P < 0.01).The brain water contents of the right basal ganglia in the ICH group were augmented(P < 0.01),they were reduced by escin(P < 0.01).The brain water contents were reduced in the Escin+IL-1? groups compared with the ICH group(P < 0.05).Evans blue extravasation in the right hemisphere was enhanced in ICH mice(P < 0.01),and it was reduced in the Escin groups(P < 0.01).Compared with the Escin group,mice in the Escin+IL-1? group showed lesser extravasation(P < 0.05).Compared with the Control group.the levels of IL-1? in the ICH group were significantly augmented(P < 0.01),the levels of IL-1? in Escin group were lower than that of the ICH group(P < 0.01).IL-1? recombinant protein reversed the effect of escin(P < 0.05)on the levels of IL-1?.The levels of Rho A and ROCK1 in ICH mice were increased(P < 0.05 or P < 0.01).Compared with the ICH group,Rho A and ROCK1 expressions were significantly decreased by escin(P < 0.05).The levels of Rho A and ROCK1 in the Escin + IL-1? group were significantly increased when compared with Escin group(P < 0.05).Compared with the Control group.the expression of I?B? in the ICH group was significantly reduced(P < 0.01).I?B? expression in Escin group was increased compared with the ICH group(P < 0.05).Co-administration IL-1? with escin decreased the expression of I?B?(P < 0.05).The expression of nuclear NF-?B in the ICH group were augmented(P < 0.01),and they were significantly attenuated by escin(P < 0.01).It was increased in the Escin + IL-1? group compared with the Escin group(P < 0.05).Mice in the ICH group showed a significant decrease in the expression of cytosolic NF-?B(P < 0.05).However,the cytosolic NF-?B levels in Escin groups were significantly increased(P < 0.05).The IL-1? recombinant protein reversed the effect of escin(P < 0.01)on the cytosolic NF-?B level.Compared with the Control group.the expressions of Occludin and Claudin-5 in the ICH group were significantly reduced(P < 0.05 or P < 0.01).Occludin and Claudin-5 expressions in Escin group were increased compared with that of the ICH group(P < 0.05).Co-administration IL-1? with escin significantly decreased the expressions of Occludin and Claudin-5(P < 0.05).Conclusion: Escin can not penetrate the blood brain barrier.Escin improves neurological outcome and the blood brain barrier function in ICH mice.The mechanism of action of escin was related to inhibiting peripheral system inflammation,reducing IL-1? level in blood circulation,regulating IL-1?/Rho A/NF-?B signaling pathway,at least,in part.Innovation: Making it clear that escin can not penetrate the blood brain barrier.Escin improves neurological outcome and the blood brain barrier function in ICH mice.The mechanism of action of escin was associated with regulating IL-1?/Rho A/NF-?B signaling pathways.