调控血小板反应蛋白-1对癫痫形成的抑制作用及机制

As the second major diseases in nervous system, epilepsy is a serious public health problem to human. The therapy of epilepsy is restricted because that the pathological mechanism of epileptogenetic is unclear by this day. A large number of studies demonstrate that abnormal synaptogenesis in brain plays an essential role in the procession of epileptogenesis and is a key to the neural network formation. Moreover, the other research found that thrombospondin-1 (TSP-1) plays a critical role in synapse formation. But if TSP-1 is involved in the progression of epileptic network formation is still unclear. . Our previous study found that the expression of TSP-1 is increased regionally in the progression of amygdaloid kindling epilepsy, accompanied with the number of synapses increased significantly. Appropriate inhibition of TSP-1 can obviously reduce the number of synapses and inhibit the epileptic process. . In order to further reveal the possible role of TSP-1 in epileptogenesis, based on our previous studies, we intend to use three kinds of representative model of epilepsy (pilocarpine, pentylenetetrazol and amygdaloid kindling animal model), revealing the role and mechanism of TSP-1 in epileptogenesis and the epileptic network formation. Further study will be done to evaluated the possibility of inhibiting epileptogenesis by regulation of TSP-1. Gene technology interference, electrophysiology, molecular biology and biochemistry techniques will be used in our study. Our findings will provide strong experimental evidence for the prevention of epileptogenesis by targeting TSP-1.

由于病理机制不清楚，癫痫防治受到制约。研究表明，癫痫是神经网络性病变，突触发生是癫痫网络形成的基础。而星形胶质细胞分泌的血小板反应蛋白-1（TSP-1）是突触形成的关键。但TSP-1在癫痫网络形成中的角色未见报道。课题组前期研究显示，杏仁核电点燃癫痫形成过程中TSP-1呈现区域性表达增强，伴随此区域突触数量显著增多。而适当抑制TSP-1能够明显减少突触数量，抑制癫痫进程。为了进一步揭示TSP-1在癫痫发生中的作用，本项目在前期研究基础上将采用多个代表性癫痫模型，利用药物/基因手段干预TSP-1功能；用分子生物学、生物化学等手段检测突触表达以及TSP-1和转化生长因子β1变化；用电生理方法分析突触后电位变化；结合行为学和脑电改变，深入揭示TSP-1在突触发生进而癫痫网络形成过程中的作用及机制，评价调控TSP-1抑制癫痫形成的可能性，为以TSP-1作为新靶点，开发特异性抗癫痫药物提供依据。

研究表明血小板反应蛋白1/转化生长因子β1（TSP-1/TGF-β1）在突触发生中起着非常关键的作用，而嘌呤2型（P2）受体对TSP-1有调节作用。但是嘌呤受体调节的TSP-1/TGF-β1在癫痫形成过程的突触发生中是否发挥作用不清楚。本研究观察了癫痫过程中TSP-1水平和突触发生的动态变化，并进一步观察了P2受体的调节作用。我们采用杏仁核电刺激癫痫模型，在癫痫形成过程中对TSP-1水平以及P2受体进行药物或者基因干预；然后评价TSP-1和TGF-β1，以及突触发生、癫痫活动的变化。研究发现，癫痫形成过程中脑内部分区域突触，尤其是兴奋性突触增生明显，并且与TSP-1和TGF-β1表达增加同步。用基因或药物手段干预，能够明显减少突触增生，抑制癫痫形成。我们结果提示P2受体介导的TSP-1/TGF-β1通路可能是癫痫形成过程中的关键因素。我们进一步实验研究发现，P2受体介导的TSP-1/TGF-β1 通路可能也参与了癫痫形成过程中的胶质增生和血管增生过程。因此，抑制这条调节通路可能是抗癫痫药物的潜在药物靶点。