经鼻给予TGF-β1对匹罗卡品诱导的癫痫持续状态大鼠海马内炎症反应的影响

Effects of Intranasal administration of TGF-β1 on hippocampus inflammation after pilocarpine induced status epilepticus

目的观察经鼻给予转化生长因子-β1(TGF-β1)对匹罗卡品致痫大鼠海马炎症反应的影响,探讨TGF-β1在癫痫持续状态后是否可以减轻炎症,及可能的抗癫痫和脑保护作用。方法 (1)采用酶联免疫吸附试验(Elisa)检测TGF-β1能否经鼻顺利入脑。(2)在匹罗卡品癫痫持续状态模型后6h对动物进行随机分为正常对照组、匹罗卡品组(Pilo组)、TGF-β1组(TGF组)。分组后对所有动物进行滴鼻,TGF组给予重组TGF-β1蛋白,Pilo组和对照组大鼠应用相同方法给予相同容量PBS溶液。各组动物于SE后1d、3d、7d处死取材,应用Elisa检测海马IL-1β、TNF-α蛋白含量,免疫组化观察海马区小胶质细胞CD11b的表达。结果 (1)经鼻滴入TGF-β1能够快速有效地到达脑组织内;(2)SE后1d、3d、7d,TGF-β1组较匹罗卡品组IL-1β、TNF-α海马内蛋白含量均有明显减少(P<0.05或P<0.01);(3)TGF-β1组和匹罗卡品组的海马区CD11b表达在1d组无明显差异,在3d和7d时间点,TGF-β1组较匹罗卡品组CD11b表达明显减少(P<0.05或P<0.01)。结论经鼻给予TGF-β1可以减少癫痫持续状态大鼠海马炎性因子IL-1β、TNF-α的表达,抑制小胶质细胞的活化,减轻海马内的炎症,在癫痫形成过程中可能具有脑保护及抗癫痫的作用。

Objective To investigate whether intranasal TGF-β1 could exert neuro-protective and anti-epileptogenic effects by modulating the inflammation in hippocampus after piocarpine induced status epilepticus.Methods Experiment1：to determine whether intranasal TGF-β1 could enter brain effectively,the concentration of TGF-β1 in the whole brain tissues was measured by ELISA.Experiment 2：six hours after pilocarpine induced status epilepticus,rats were randomly divided into control group,pilocarpine group（Pilo group）and pilocarpine＋TGF-β1 group（TGF group）.Then,rats of TGF group were intranasal administrated recombinant human TGF-β1,while rats of Pilo-group and control group were treated identically with the same volume of PBS.1d,3d,7d after SE,the expression of CD11b in hippocampus was detected by immunohistochemistry,and the IL-1β and TNF-α level in the hippocampus was measured by ELISA individually.Results（1）Intranasal TGF-β1 could effectively enter brain rapidly.（2）After intranasal administration of TGF-β1,concentration of IL-1β and TNF-α in the hippocampus in pilocarpine treated rats was significantly decreased at 1d,3d,7d after SE（P0.05 or P0.01）.（3）Expression of CD11b in the hippocampus of was no significant difference between TGF-group and Pilo-group at 1d after SE.However,at 3d and 7d after SE,it was significant reduced in the TGF-group comparing with Pilo-group（P0.05 or P0.01）.Conclusion Intranasal TGF-β1 can inhibit the activation of microglia and reduce cytokines IL-1β and TNF-αconcentration in the hippocampus of rats,attenuate the inflammation of hippocampus,may have a potent neuro-protective and anti-epileptogenic role in epileptogenesis.