GAP-43和Sema3A蛋白在戊四氮点燃慢性癫痫大鼠海马的表达

The expression of GAP-43 and Sema3A in the chronic epileptic rats' hippocampus which was kindled by pentylenetetrazol

目的:探讨神经生长相关蛋白43(growth-associated protein 43,GAP-43)和semaphorin3A(Sema3A)蛋白在慢性癫痫模型大鼠海马的表达。方法:将成年雄性SD大鼠41只随机分为正常对照组和癫痫组,通过腹腔注射戊四氮(40 mg/kg)诱导慢性癫痫模型,取大脑海马组织,应用免疫组织化学和蛋白免疫印迹分析(Western Blot)方法,观察GAP-43蛋白及Sema3A蛋白在大鼠海马各区的表达变化。结果:免疫组织化学结果显示:GAP-43免疫阳性颗粒主要位于阳性细胞的胞浆和细胞膜表面,胞浆和突起被染成深棕色,靠近轴突一侧的胞浆染色较深;Sema3A免疫阳性颗粒主要位于阳性细胞的胞浆中,分布比较均匀,胞浆被染为棕黄色,染色较浅。在海马的齿状回及CA3区,癫痫组大鼠GAP-43蛋白的表达高于对照组大鼠,而Sema3A蛋白的表达则低于对照组,差异有统计学意义(P<0.05);在海马的CA1区,癫痫组大鼠GAP-43和Sema3A蛋白的表达均显著高于对照组大鼠,差异具有统计学意义(P<0.01)。Western Blot结果显示:在整个海马组织内GAP-43蛋白在癫痫组的表达为0.45±0.21,明显高于对照组(0.28±0.23),差异具有统计学意义(P<0.05);Sema3A蛋白在癫痫组和对照组的表达分别为1.18±0.55和1.55±1.96,虽有差异,但并无统计学意义(P>0.05)。结论:GAP-43蛋白在癫痫大鼠海马中表达升高,Sema3A在癫痫大鼠海马中具有区域性差异表达;这两种蛋白的表达变化可能参与了癫痫的发病机制。

Objective： Investigating the expression of GAP- 43 （ growth-associated protein 43 ） and Sema3A （ semaphorin 3A） in the chronic epileptic rats＇ hippocampus. Method： 41 male rats were divided into control group and epileptic group （EP group） randomly. The chronic epileptic rats models were established by injecting pentylenetetrazol intraperitoneally （PTZ, 40 mg/kg）. Then take out the hippocampus of brain to observe the expression change of GAP-43 and Sema3A in every region of hippocampus by using the method of immunohistochemistry and Western Blot. Results： The resuit of immunohistochemistry showed that GAP- 43 immunoreactive particles were mainly in the cytoplasm of positive ceils and on the surface cell membrane. So that the cytoplasm and axon were dyed into clark brown, and near the ceil-substance side cytoplasm was dyed deeper. Sema3A particles were mainly distributed in the cytoplasm of positive ceils uniformly, and the cytoplasm was dyed into lighter brown. In the dentate gyrus and CA3 region of hippocampus, the expression of GAP- 43 protein in EP group was higher than control group, but the expression of Sema3A protein in EP group was lower than control group, there was a statistically significance compared EP group with control group （ P 〈 0.05 ）. In the CA1 region of hippocampus, the expression of GAP-43 and Sema3A protein in EP group was significantly higher than control group, the difference was a statistically significance （P 〈 0.01 ）. Western Blot result showed that the expression of GAP-43 protein in hippocampus of EP group （0.45 ± 0.21 ） was significantly higher than control group （0.28 ± 0. 23）, but difference was no statistically significance （P 〈 0.05 ）. The expression of Sema3A protein in hippocampus of EP group （ 1.18±0.55） was lower than control group （ 1.55 ± 1.96）, also there was a difference, no statistically significance （ P 〉 0.05 ）. Conehtsion： The expression of GAP- 43 protein in hippocampus of EP group was higher. The expression of Sema3A was regional difference in the hippocampus of EP group. The expression changes of GAP- 43 and Sema3A may be involved in the pathogenesis of epilepsy.