乙酰唑胺对慢性癫痫大鼠海马NF-κB p65、IL-1β、IL-6及TNF-α表达的影响  

Effect on the expression of NF-κB p65、IL-1β、IL-6 and TNF-α in the hippocampus of chronic epilepsy rats by acetazolamide

作　　者：余涵[1,2,3] 陈丽[1] 马猛[1] 周玉波[1] 王林杰[1] 罗亚楠[1] 朱晓琴[1] 雷水生[4] 

机构地区：[1]广州医科大学生理教研室,广东广州511436 [2]湖北医药学院襄阳医院 [3]襄阳市第一人民医院病理科,湖北襄阳441000 [4]广州医科大学附属第五医院皮肤科,广东广州510700

出　　处：《中风与神经疾病杂志》2015年第8期730-733,共4页Journal of Apoplexy and Nervous Diseases

基　　金：广州市科技计划项目(No.11C32020697;No.2013J4100103);广州市属高校科研计划项目(No.2012c128;No.2012D001);广东省省级科技计划项目(No.2013B02180091)

摘　　要：目的观察戊四氮(Pentylenetetrazole,PTZ)致慢性癫痫大鼠发作以及应用AQP4抑制剂乙酰唑胺(Acetazolamide,AZA)干预后对海马核因子-κB p65(NF-κB p65)、白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的表达影响,探讨AQP4与炎症因子表达间的关系及乙酰唑胺抑制癫痫发作的可能机制。方法将50只健康成年SD大鼠随机分为对照组(每日腹腔注射生理盐水)、致痫组(每日腹腔注射亚惊厥剂量PTZ35 mg/(kg·d)建立慢性癫痫大鼠模型)、乙酰唑胺干预组[每日先腹腔注射AZA35 mg/(kg·d)预处理,30 min后再腹腔注射亚惊厥剂量PTZ]。连续注射28 d,每天记录大鼠的发作级别。最后一次给药1 d后处死所有大鼠以备实验,RT-q PCR和ELISA检测大鼠海马中IL-1β、IL-6、TNF-α的表达;Western blot检测海马内NF-κB p65表达。结果对照组无明显癫痫发作,RT-q PCR和ELISA检测结果显示致痫组IL-1β、IL-6、TNF-α水平均显著高于对照组(P<0.001);乙酰唑胺干预组的三者水平明显低于致痫组(P<0.05),但仍高于对照组(P<0.05);Western blot结果显示致痫组海马组织NF-κB p65的表达与对照组相比显著升高(P<0.01),而乙酰唑胺干预组表达量与致痫组相比明显下降(P<0.05),但仍高于对照组(P<0.05)。结论乙酰唑胺抑制癫痫的发作的作用机制可能与乙酰唑胺抑制星形胶质细胞膜上AQP4的表达、改善星形胶质细胞水肿损伤状况、下调炎性因子表达有关。Objective To observe the effect of AQP4 inhibitors acetazolamide on change of behavior and expression of NF-κB p65,IL-1β,IL-6,TNF-α in hippocampus and serum of PTZ induced chronic epileptic rats. Then explore the relationship between the AQP4 expression with pro-inflammatory factors and the possible antiepileptic mechanism of acetazolamide. Methods The experimental rats were randomly divided into three groups： control group,in which the rats were daily intraperitoneal injection of normal saline; epileptic group,in which the rats were daily administered subconvulsive dose of PTZ（ 35 mg / kg）. Acetazolamide intervention group,in which the rats were daily treated with AZA（ 35 mg / kg） 30 min before PTZ administering. All groups were administered for 28 consecutive days and recorded degree of daily seizure. All rats were sacrificed for experiments one day after the last administration of each drug. Real-time quantitative PCR（ RT-q PCR）and ELISA were used to observe the expression of IL-1β、IL-6 and TNF-α in the hippocampus. Western blot were used to examine the expression of the NF-κB p65 in hippocampus. Results Behavioral observation showed that rats in control group had no obvious epileptic seizure. RT-q PCR and ELISA showed PTZ group and the acetazolamide group IL-1β,IL-6,TNF-α levels were significantly higher than control group（ P〈0. 001）. Meanwhile,the levels of pro-inflammatory cytokines in the AZA group was significantly lower than that of PTZ group（ P〈0. 05）,but still higher than the control group（ P〈0. 05）. Western blot results showed that NF-κB p65 in PTZ group was significantly higher compared to the control group（ P〈0. 01）,and acetazolamide group was lower compared with PTZ group（ P〈0. 05）,but still higher than that of control group（ P〈0. 05）. Conclusion The antiepileptic mechanism of acetazolamide may be suppressing AQP4 expression of astrocytes、improving astrocytes edema anddownregulating pro-inflammatory cytokine expression.

关 键 词：慢性癫痫 乙酰唑胺 水通道蛋白4 炎症因子 NF-ΚB P65 

分 类 号：R742.1[医药卫生—神经病学与精神病学]