外源性硫化氢处理对癫痫致海马神经元损伤的影响  

Effect of exogenous hydrogen sulfide on hippocampal neuronal damage induced by epilepsy

作　　者：彭昌[1] 罗孝美[2] 幸黔鲁[1] 刘颖[1] 谭梅[1] 

机构地区：[1]遵义医学院附属医院儿内科,遵义563000 [2]遵义医学院生理学教研室,遵义563000

出　　处：《中华神经医学杂志》2015年第8期805-809,共5页Chinese Journal of Neuromedicine

基　　金：遵义医学院硕士启动基金[院字（2009）19号]

摘　　要：目的探讨给予外源性硫化氢（H2S）供体硫氢化钠（NariS）对氯化锂一匹罗卡品致癫痫持续状态幼鼠海马神经元损伤的影响。方法3~4周龄SD大鼠按随机数字表法分为5组：正常组、正常＋NaHS组、癫痫组、癫痫＋NaHS组、癫痫＋羟胺组。后3组大鼠应用腹腔注射氯化锂＋阿托品溶液＋盐酸匹罗卡品溶液方法建立癫痫持续状态模型，并分别给予NariS10mg／kg或羟胺1mg／kg或等量生理盐水腹腔注射。干预后7d各组取6只大鼠收集海马组织及血浆，采用分光光度法测定血浆及海马H：S浓度，采用RT-PCR检测海马胱硫醚β-合酶（CBS）mRNA表达水平，采用Nissl染色观察海马神经元形态，采用Westemblotting检测海马组织神经元特异性烯醇化酶（NSE）蛋白表达水平。结果分光光度法和RT—PCR检测结果显示：血浆和海马H2S浓度及CBSmRNA表达水平在癫痫组均明显低于正常组，在癫痫-NaHS组均显著高于癫痫组，而在癫痫＋羟胺组均显著低于癫痫组，差异均有统计学意义（P〈0．05）。Nissl染色结果显示：癫痫组海马CA3区尼氏体明显减少．锥体细胞排列紊乱．而癫痫＋NaHS组海马尼氏体缺失及锥体细胞排列紊乱较癫痫组明显减轻。Westemblotting结果显示：海马组织中NSE表达水平在癫痫组显著高于正常组，在癫痫＋NaHS组显著低于癫痫组，而在癫痫十羟胺组显著高于癫痫组，差异均有统计学意义（氏0．05）。结论给予外源性H-S供体NariS能够显著上调海马组织CBSmRNA表达水平，提高血浆及海马组织H2S含量，降低海马NSE表达水平，从而改善癫痫所致的海马神经元损伤。Objective To explore the effects of exogenous supplier of hydrogen sulfide （H2S） sodium hydrosulfide （NariS） on hippocampal neuronal damage in immature rats with status epilepticus （SE） induced by lithium-pilocarpine. Methods Three or four weeks old Sprague Dawley rats were divided into five groups according to random number table： normal group, normal＋NariS group, epilepsy group, epilepsy＋NariS group and epilepsy＋hydroxylamine （HA） group （n=6）. SE models were induced by lithium-pilocarpine in immature rats of epilepsy group, epilepsy＋NariS group and epilepsy＋HA group; meanwhile, NariS （10 mg/kg） or hydroxylamine （1 mg/kg） or equivalent normal saline were given by intraperitoneal injection, respectively. Hippocampus tissues and plasma were collected, and the concentrations of H2S in the plasma and hippocampus were detected by spectrophotometer. The cystathionine beta synthase （CBS） mRNA expression in the hippocampus was assayed by real-time PCR. Hippocampal neurons were observed by Nissl staining. The neuron-specific enolase （NSE） expression in the hippocampus was assayed by Western blotting. Results Spectrophotometer and real-time PCR results showed that the concentration of H2S in the plasma and hippocampus and the CBS mRNA levels in the hippocampus of the epilepsy group were significantly lower than those in the normal group （P〈0.05）;these indexes in the epilepsy＋NariS group were up-regulated as compared with those in the epilepsy group, while those in the epilepsy＋HA group were significantly lower than those in the epilepsy group （/9〈0.05）. Nissl staining showed that decreased number of nissl bodies and disordered arrangement of pyramidal cells in the hippocampus CA3 area of epilepsy group were noted as compared with those in the normal group; the loss of nissl bodies and the disordered arrangement of pyramidal cells in the hippocampus in epilepsy＋NariS group were improved as compared with those in the epilepsy group. Western blotting showed th

关 键 词：硫化氢 癫痫持续状态 海马 神经元 

分 类 号：R742.1[医药卫生—神经病学与精神病学]