麝香黄芪复方滴丸对缺血再灌注损伤大鼠脑组织紧密连接蛋白表达影响

Effect of Compound Shexiang Huangqi Dropping Pill(麝香黄芪复方滴丸)on Expressions of Tight Junction Proteins in Brain Tissue of Rat with Ischemia Reperfusion Injury

目的旨在通过麝香黄芪复方滴丸对脑缺血大鼠模型紧密连接蛋白表达影响,探讨麝香黄芪复方滴丸对缺血再灌注大鼠血脑屏障修复的作用机制。方法将440只大鼠随机分成假手术组、模型组、麝香黄芪复方滴丸组、依达拉奉组,每组110只。利用改良线栓法制备大鼠大脑中动脉栓塞缺血再灌注模型,在造模前3 d,假手术组、模型组按照15 mL/kg灌胃生理盐水,麝香黄芪复方滴丸组及依达拉奉组分别按15 mL/kg给药,1次/d。造模后继续按此标准给药至取材日期。采用Western Blot测定缺血脑组织中Occludin、Claudin-5、JAM-A及ZO-1蛋白表达。结果(1)与假手术组相比,模型组缺血脑组织Occludin蛋白、Claudin-5蛋白、JAM-A及ZO-1蛋白表达水平明显下降,差异有统计学意义(P<0.05)。(2)与模型组相比,麝香黄芪复方滴丸组和依达拉奉组缺血脑组织Occludin蛋白、Claudin-5蛋白、JAM-A及ZO-1蛋白表达水平均有上升,差异有统计学意义(P<0.05)。(3)在造模后的1 d,依达拉奉组缺血脑组织Claudin-5蛋白表达水平上升较麝香黄芪复方滴丸组明显,在造模后的第3、5天,麝香黄芪复方滴丸组缺血脑组织Claudin-5蛋白表达水平上升均高于依达拉奉组,但差异无统计学意义(P>0.05);在各治疗组中Occludin蛋白、JAM-A及JAM-A蛋白均有升高,麝香黄芪复方滴丸组升高明显,但差异无统计学意义(P>0.05)。结论麝香黄芪复方滴丸对血脑屏障的修复具有协同作用,可能是通过上调缺血脑组织中Occludin、Claudin-5、JAM-A、ZO-1的蛋白表达来维持血脑屏障紧密连接的完整性,实现脑保护、降低脑水肿的作用。

Objective To investigate the mechanism of repairing blood-brain barrier in cerebral ischemia reperfusion injury rats by the effect of Compound Shexiang Huangqi Dropping Pill(麝香黄芪复方滴丸)on the expressions of tight junction proteins in cerebral ischemia rat model.Methods A total of 440 rats were randomly divided into sham operation group,model group,Compound Shexiang Huangqi Dropping Pill group and edaravone group,with 110 rats in each group.Three days before modeling,the sham operation group and the model group were given normal saline by intragastric administration with the dose of 15 mL/kg,and the Compound Shexiang Huangqi Dropping Pill group and the edaravone group were given the corresponding drugs with the dose of 15 mL/kg,once a day,respectively.After modeling,the drug was administered according to this standard until the day of extraction.The protein expressions of Occludin,Claudin-5,JAM-A and ZO-1 in ischemic brain tissue were determined by Western Blot.Results(1)Compared with those of the sham operation group,the expression levels of Occludin,Claudin-5,JAM-A and ZO-1 in ischemic brain tissue in the model group were significantly decreased with significant difference(P<0.05).(2)Compared with those of the model group,the expression levels of Occludin,Claudin-5,JAM-A and ZO-1 in ischemic brain tissue in the Compound Shexiang Huangqi Dropping Pill group and edaravone group were increased and the differences were significant(P<0.05).(3)On the first day after modeling,the protein expression level of Claudin-5 in ischemic brain tissue in the edaravone group was significantly higher than that in the edaravone group.On the third and fifth day after modeling,the protein expression level of Claudin-5 in the Compound Shexiang Huangqi Dropping Pill group was higher than that in the edaravone group,but the difference was not significant(P>0.05).The expressions of Occludin,JAM-A and ZO-1 were increased in all treatment groups,and the increase was significant in the Compound Shexiang Huangqi Dropping Pill group,but the difference was not significant(P>0.05).Conclusions Compound Shexiang Huangqi Dropping Pill has a synergistic effect on the repair of blood-brain barrier,which may be to maintain the integrity of the tight junction of blood-brain barrier,realize brain protection and reduce brain edema by up-regulating the protein expressions of Occludin,Claudin-5,JAM-A and ZO-1 in ischemic brain tissue.