目的 观察盐酸小檗碱(BBH)预处理对青霉素钠诱发小鼠癫痫发作程度、发作潜伏期的影响,并探讨其机制.方法 雄性昆明种小鼠50只,适应性饲养3 d后随机分为生理盐水组、模型组、丙戊酸钠(VPA)组、BBH1组、BBH2组.VPA组腹腔注射VPA 40 mg/kg,BBH1组以BBH 25 mg/kg灌胃,BBH2组以BBH 50 mg/kg灌胃,生理盐水组和模型组分别以等量生理盐水和β-环糊精灌胃.各组每天给药1次,连续给药3 d.至末次给药次日,除生理盐水组外,其余四组均腹腔注射青霉素钠700万IU/kg诱发癫痫.记录各组小鼠从给药到出现癫痫症状的潜伏期.根据Racine分级标准评估癫痫发作分级,记录癫痫大发作(>Ⅲ级)发生情况.发作持续1 h后,颈椎脱臼处死动物并断头取脑,采用液相色谱-质谱联用法(LC-MS)检测海马组织中的谷氨酸(Glu)、γ-氨基丁酸(GABA)、五羟色胺(5-HT).结果 模型组癫痫发作Ⅲ级2只、Ⅳ级7只、Ⅴ级1只,癫痫大发作8只;VPA组Ⅲ级6只、Ⅳ级3只、Ⅴ级1只,共4只达到大发作;BBH1组Ⅲ级3只、Ⅳ级6只、Ⅴ级1只,7只大发作;BBH2组Ⅱ级1只、Ⅲ级6只、Ⅳ级3只、Ⅴ级1只,癫痫大发作4只.BBH2组小鼠癫痫发作达到Ⅲ级以上的数量与模型组相比,P均<0.05.模型组、VPA组、BBH1组、BBH2组癫痫发作潜伏期分别为(142.4±68.4)、(237.5±132.6)、(246.9±59.8)、(260.5±52.4)s,BBH1组、BBH2组癫痫发作潜伏期较模型组延长(P均<0.05).VPA组海马组织中GABA表达水平高于模型组(P<0.05).模型组Glu/GABA高于生理盐水组(P<0.05);VPA组Glu/GABA低于模型组(P<0.05);BBH1组、BBH2组Glu/GABA低于模型组,但差异无统计学意义.BB1组Glu/5-HT低于模型组(P<0.05).结论 BBH预处理有助于降低小鼠癫痫发作分级,延长癫痫发作潜伏期,其中50 mg/kg剂量效果优于25 mg/kg剂量;BBH的抗癫痫机制可能与调节海马组织内神经递质水平有关.%Objective To observe the effects of berberine (BBH) pretreatment on the degree and the incubation period of epileptic seizure induced by penicillin in mice and to discuss the mechanism.Methods After adaptive feeding of 3 days,fifty male Kunming mice were randomly divided into five groups including the saline group,model group,VPA group,BBH 1 group,and BBH 2 group.The mice in the VPA group were given intraperitoneal injection of 40 mg/kg VPA,mice in the BBH 1 group were given 25 mg/kg BBH by gavage and BBH 2 group with 50 mg/kg BBH in the same way.The saline and model groups were given the same volume of saline or 25% β-cyclodextrin,and all the groups were administered once a day for three days.Up to the next day of the last time,except for the saline group,all the other four groups were given an intraperitoneal injection of 7×106 IU/kg penicillin to induce epileptic seizure.We recorded the incubation period for each group of mice from administration to the onset of epilepsy.We evaluated the onset classification byRacine criterion and recorded the seizure attack (more than III-grade).Animals showing seizure attack for 1 h were killed by dislocation of cervical vertebra.Brain tissues were taken out and processed for further estimation,we used LC-MS to test the content of glutamic acid (Glu),γ-aminobutyric acid (GABA) and serotonin (5-HT).Results As for the seizures,there were 2 cases of class Ⅲ,7 cases of class Ⅳ,1 case of class V,and 8 cases of grand mal epilepsy,respectively,in the model group;in the VPA group,they were 6,3,1,and 4;in the BBH 1 group,they were 3,6,1,and 7;in the BBH 2 group,there was i case of class II,6 cases of class Ⅲ,3 cases of class Ⅳ,1 case of class V,and 4 cases of grand mal epilepsy.The number of mice with seizures more than class Ⅲ in the BBH 2 group was significantly different as compared with that of the model group (all P<0.05).The incubation period of the model group,VPA group,BBH 1 and BBH 2 groups were (142.4±68.4),(237.5±132.6),(246.9±59.8),and (260.5±52.4)s.The incubation period of the BBH 1 and BBH 2 groups was longer than that of the model group (all P<0.05).The level of GABA in the VPA group was higher than that of the model group (P<0.05).The Glu/GABA of the model group was higher than that of the saline group (P<0.05).The Glu/GABA of the VPA group was lower than that of the model group (P<0.05).The levels of Glu/GABA in the BBH1 group and the BBH2 group were lower than that of the model group,but the difference was not statistically significant.The Glu/5-HT in the BBH1 group was lower than that of the model group (P<0.05).Conclusions The BBH pretreatment helps to reduce the scale of epileptic seizures in mice and prolong the incubation period.Among them,the overcome of 50 mg/kg dose is better than 25 mg/kg.The anti-epileptic mechanism of BBH may be associated with regulating neurotransmitter levels in the hippocampus.
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