L-NNA与L-Arg对大鼠KA诱导性癫痫发作的影响及其细胞学基础与机制探讨
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摘要
癫痫多发、难治、危害性大，复杂部分性癫痫发作为其典型代表之一，深入探讨癫痫发作可能的细胞学基础与分子机制，已成为最终认识和解决癫痫问题的关键所在。本研究采用公认的复杂部分性癫痫发作模型——大鼠红藻氨酸诱导性癫痫发作，以组成性一氧化氮合酶抑制剂L-硝基精氨酸及一组小剂量的L-精氨酸予以干预，用多种定性、定量或半定量方法，从行为学评估(制备录像资料)、形态学观察(免疫组化与Nissl复染)以及生物化学(比色分析)与分子生物学检验(Western印迹分析与RT-PCR)的不同角度和层次，对复杂部分性癫痫发作中一氧化氮介质途径的作用及其可能的细胞学基础与生化机制做了观察和探讨。结果发现：红藻氨酸(10mg／kg体重，i．p．)可使95％以上的动物(年轻、成年、雄性)发生一组有显著时间相关性的复杂部分性癫痫发作行为，这组发作行为伴有受累脑区特别是海马结构中Fos-、IL-6-、GFAP-及OX42—免疫反应性的快速增强及NO_2~-／NO_3~-含量的迅速升高。在发作数小时及数日后，受累脑区内还出现Hsp72／73—免疫反应性的增强及诱导型NOS mRNA。在基础状态下，L-硝基精氨酸(50mg／kg体重，i．p．)与小剂量的L-精氨酸(10、40及160mg／kg体重，i．p．)慢性给药(2／日，共给药9次)未引起明显的行为反应，但在红藻氨酸(10mg／kg体重，i．p．)给药后，动物的发作行为都发生了显著变化。L-硝基精
To further determine whether or not nitric oxide mediator (s) is involved in epilepsy and address the cellular and biochemical mechanisms, we evaluated the effect of L-nitroarginine (50mg/kg b. w. , i. p. , twice daily for 9 times) or L-arginime (10, 40, or 160mg/kg b. w. , i. p. , twice daily for 9 times) on kainic acid (10mg/kg b. w. , i. p. ) -induced seizures in young adult male rats and the corresponding changes in Fos-, IL-6-, Hsp 72/73-, GFAP-, and OX42-im-munoreactivity (ir) in the most affected brain region, the hippocampal formation. It was found that kainic acid caused in almost all of the animals a series of stereotype behavorial changes, and corresponding rapid increases in Fos-, IL-6-, GFAP-, and OX42-ir and NO_2~-/NO_3~- concentration in the hippocampal formation. Hsp72/73-ir and the mRNA of inducible nitric oxide synthase were also found increased several hours to days later. Under basal conditions, the chronic treatment of L-nitroarginine or L-arginine caused no apparent behavioral changes. However, the chronic pretreatment of L-nitroarginine so dramatically promoted and enhanced the kainic acid-induced behavioral seizures that almost all of the animals died at about 3 hours after kainic acid injection. In contrast, the chronic pretreatment of L-arginine at 40 or 160mg/kg b. w. dose markedly
To further determine whether or not nitric oxide mediator (s) is involved in epilepsy and address the cellular and biochemical mechanisms, we evaluated the effect of L-nitroarginine (50mg/kg b. w. , i. p. , twice daily for 9 times) or L-arginime (10, 40, or 160mg/kg b. w. , i. p. , twice daily for 9 times) on kainic acid (10mg/kg b. w. , i. p. ) -induced seizures in young adult male rats and the corresponding changes in Fos-, IL-6-, Hsp 72/73-, GFAP-, and OX42-im-munoreactivity (ir) in the most affected brain region, the hippocampal formation. It was found that kainic acid caused in almost all of the animals a series of stereotype behavorial changes, and corresponding rapid increases in Fos-, IL-6-, GFAP-, and OX42-ir and NO_2~-/NO_3~- concentration in the hippocampal formation. Hsp72/73-ir and the mRNA of inducible nitric oxide synthase were also found increased several hours to days later. Under basal conditions, the chronic treatment of L-nitroarginine or L-arginine caused no apparent behavioral changes. However, the chronic pretreatment of L-nitroarginine so dramatically promoted and enhanced the kainic acid-induced behavioral seizures that almost all of the animals died at about 3 hours after kainic acid injection. In contrast, the chronic pretreatment of L-arginine at 40 or 160mg/kg b. w. dose markedly
引文
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