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目的:动态观察小分子GTPase Rho家族的Rnd1 mRNA及其蛋白在氯化锂-毛果芸香碱(匹罗卡品)致大鼠模型海马中的表达变化,探讨其在颞叶癫发生发展中的作用。方法:在氯化锂-毛果芸香碱颞叶癫模型中应用逆转录聚合酶链反应(RT-PCR)检测癫持续发作(SE)后各时间点海马内Rnd1 mRNA的表达变化,并运用免疫组织化学染色法及Neo-Timm染色法分别检测齿状回门区、CA1区及CA3区中该蛋白在不同时间点的表达变化及苔藓纤维出芽(MFS)情况。结果:实验发现模型组于SE后8h内即出现Rnd1表达上调,SE后约1d达高峰,7d左右回复至对照组水平,此后其mRNA表达水平与对照组相似;而免疫组化染色发现Rnd1蛋白表达从SE后8h内即开始上调,约3d达高峰,至7d虽略有回落,但仍高于对照组水平,且这种情况可一直持续至慢性期。结论:急性期海马齿状回门区Rnd1表达上调可能通过促进MFS的发生参与了颞叶癫的发生。
OBJECTIVE: To observe the expression of Rnd1 mRNA and protein of small molecule GTPase Rho family in hippocampus of rat induced by lithium chloride-pilocarpine (pilocarpine), and to explore its role in the development of temporal lobe epilepsy . Methods: RT-PCR was used to detect the expression of Rnd1 mRNA in the hippocampus of the lithium-pilocarpine temporal lobe epilepsy model at each time point after the continuous epileptic seizure (SE) The expression changes of the protein in the dentate gyrus, CA1 and CA3 regions at different time points and the moss fiber sprouting (MFS) were detected by chemical staining and Neo-Timm staining. Results: In the experimental group, the expression of Rnd1 was up-regulated within 8 hours after SE in rats. The expression of Rnd1 peaked at about 1 day after SE and returned to the level of control after 7 days. The expression of Rnd1 protein was similar to that of control group The expression began to rise within 8 h after SE, and peaked at about 3d. Although it dropped slightly to 7d, it was still higher than that of the control group, and this condition continued until the chronic phase. Conclusion: Upregulation of Rnd1 in the dentate gyrus in the hippocampus may be involved in the development of temporal lobe epilepsy by promoting the occurrence of MFS.