TLR9、MyD88在颞叶癫痫大鼠海马组织中表达的动态变化

Dynamic expression changes of TLR9 and MyD88 in hippocampus of temporal lobe epilepsy rats

目的观察氯化锂-匹罗卡品致痫大鼠各期海马中Toll-样受体9(TLR9)、髓样分化因子(MyD88)表达的变化,探讨其是否与颞叶癫痫发生有关。方法 SD雄性大鼠120只,随机分为对照组(30只)和模型组(90只),腹腔注射氯化锂。18 h^20 h后模型组腹腔注射匹罗卡品诱导癫痫持续状态(SE);对照组予等量生理盐水取代匹罗卡品腹腔注射。对照组和造模成功的模型组依据腹腔注射后时间随机分为10个亚组:急性模型组(SE后3 h、6 h、9 h、12 h、1 d、3 d、7 d);潜伏模型组(SE后14 d、28 d);慢自发发作组(SE后56 d)。每亚组动物模型组9只,对照组3只。免疫组化、蛋白印迹、RT-PCR技术测定各亚组癫痫大鼠海马内TLR9、MyD88的表达。结果TLR9、MyD88在模型组海马内表达明显增多,与对照组相比,差异有显著性(P<0.05)。模型亚组内,TLR9、MyD88在急性期和慢性期表达明显增高,而潜伏期无明显表达变化。其中急性期内的增高多集中在癫痫发作后6 h;3组比较差异有显著性(P<0.05)。结论大鼠海马内TLR9、MyD88表达增多可能与颞叶癫痫发病有关,探讨其机制可能为颞叶癫痫的治疗提供新的靶点。

Objective To investig ate the dynamic expression changes of TLR9 and MyD88 in hippocampus of the epilepsy rats induced by from lithium chloride-pilocarpine during the development of epilepsy,and to explore whether they induced the pathogenisis of the temporal lobe epilepsy.Methods 120 male healthy Sprague-Dawley rats were divided randomly into control group and Li Cl-Pilocarpine experimental group.The experimental rats were intraperitoneally injected with Li Cl（127 mg/kg） and Pilocarpine（30 mg/kg） to induce status epilepticus（SE）.SE was stopped with 10% chloral hydrate intraperitoneally injecting 60 minutes later.The rats in the control group and the experimental group were randomly divided into 10 subgroups：acute group（3 h,6 h,9 h,12 h,1 d,3 d,7 d）,latent group（2 w,4 w）,chronic group（8 w）.Survival rats were continuously observed for onset and recurrence of spontaneous seizures every day and were grouped and sacrificed at corresponding time after the last pilocarpine injection respectively.The control rats were mitted tales doses of 0.9% saline and grouped and sacrificed at the same time after the saline injection.And all groups were executed by using immunohistochemistry,Western-blotting and RT-PCR analysis to examine the dynamic expression of TLR9 and MyD88 in the subfields of each gourps hippocampus.Results Compared with control group,the expression of TLR9 and MyD88 increased in all of the model group（P〈0.05）.In all of model group,the expression of TLR9 and MyD88 increased more significantly in acute stage and chronic stage than the latent stage（P〈0.05）.Acute phase increase more concentrated in 6 h.Conclusion The evelated expressions of TLR9 and MyD88 in epilepsy rat hippocampus indicate they could be responsible for the epileptogensis of temporal lobe epilepsy.The TLR9/MyD88 pathway may play an important role in the pathogenisis of temporal lobe epilepsy.