银杏内酯B对癫痫大鼠血脑屏障保护作用的研究  

Protective effect of Ginkgolide B on the blood-brain barrier in epileptic rats

作　　者：鲁光辉[1] 李新峰 冯亮 罗玲[1] 王海华[1] U Guanghui;LI Xinfeng;FENG Liang;LUO Ling;WANG Haihua(Handan Second Hospital,Handan 056000,China)

机构地区：[1]邯郸市第二医院,河北邯郸056000

出　　处：《药物评价研究》2021年第10期2176-2182,共7页Drug Evaluation Research

基　　金：河北省医学科学研究课题计划(20191789)。

摘　　要：目的探讨银杏内酯B(GB)对癫痫模型大鼠血脑屏障(BBB)通透性的保护作用及机制。方法将180只雄性SD大鼠按随机数字表法分为对照组(生理盐水)、模型组(生理盐水)、丙戊酸钠(VPA,300 mg/kg)组和GB低、中、高剂量(2.5、5.0、10.0 mg/kg)组,每组30只。采用氯化锂-匹罗卡品诱导制备癫痫大鼠模型;sc匹罗卡品前1 h,各组分别ip给药。观察各组大鼠癫痫发作潜伏期及持续时间;干湿比质量法测定脑组织含水量,伊文思蓝渗透法测定BBB通透性;黄嘌呤氧化法和钼酸铵法分别检测脑组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力,硫代巴比妥酸法检测丙二醛(MDA)含量,酶联免疫吸附法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)含量,Western blotting检测闭合蛋白(Occludin)、闭锁连接蛋白-1(ZO-1)、脑组织基质金属蛋白酶⁃2(MMP⁃2)、脑组织基质金属蛋白酶-9(MMP⁃9)蛋白表达。结果与模型组比较,VPA组和GB中、高剂量组大鼠癫痫发作潜伏期显著延长、发作持续时间缩短(P<0.01);脑组织含水量和伊文思蓝渗透量显著降低(P<0.05、0.01);脑组织SOD和CAT活性显著升高且MDA、TNF-α、IL-1β、IL-6含量显著降低(P<0.05、0.01);Occludin、ZO-1表达显著上调且MMP⁃2、MMP⁃9表达显著下调(P<0.01);且GB高剂量组作用优于VPA组。结论GB对癫痫大鼠BBB通透性具有保护作用,其机制可能与抑制氧化应激和炎症,以及上调Occludin、ZO-1表达并下调MMP⁃2、MMP⁃9表达有关。Objective To investigate the protective effect of ginkgolide B(GB)on the permeability of blood-brain barrier(BBB)in rats with epilepticus(EP)and its mechanism.Methods Totally 180 male SD rats were randomly divided into normal control group(normal saline),model group(normal saline),sodium valproate(VPA)group(300 mg/kg)and GB low,medium and high dose groups(2.5,5.0,10.0 mg/kg).The EP rat models were pruducted by induction with lithium chloride and pilocarpine;1h before pilocarpine injection,the drugs were given to rats in each group by administered intraperitoneally.The behavior of rats in each group was observed;the water content of brain tissue was detected by wet and dry specific gravity method,and the permeability of BBB was detected by Evans blue penetration method.The activity of SOD and CAT were detected by xanthine oxidation method and ammonium molybdate method,and the MDA content was detected by thiobarbituric acid method;the content of TNF-α,IL-1β,IL-6 were detected by ELISA.The expression of Occludin,ZO-1,MMP⁃2,MMP⁃9 were detected by Western blotting method.Results Compared with model group,the number of seizures of the rats in VPA group and GB medium,high dose groups was reduced,the seizure duration was shortened(P<0.01).The brain tissue water content and Evans blue penetration were reduced(P<0.05 or 0.01).The activity of SOD,CAT in brain tissue were increased and MDA,TNF-α,IL-1β,IL-6 content were decreased(P<0.05 or 0.01);the expression of Occludin,ZO-1 were up-regulated and MMP⁃2,MMP⁃9 expression were down-regulated(P<0.01).Compared with VPA group,the number of seizures of the rats in GB high dose group was reduced,and the seizure duration was shortened(P<0.05 or 0.01).The Evans blue penetration were reduced(P<0.05).The activity of SOD in brain tissue were increased and MDA,TNF-α,IL-6 content were decreased(P<0.05 or 0.01);the expression of Occludin,ZO-1 were up-regulated and MMP⁃2,MMP⁃9 were down-regulated(P<0.05 or 0.01).Conclusion GB has protective effect on the BBB permeability of

关 键 词：银杏内酯B 癫痫 血脑屏障 氧化应激 炎症 

分 类 号：R285.5[医药卫生—中药学]