新型硫化氢供体调控P2Y12受体促进小胶质细胞极化抑制癫痫的分子机制研究

癫痫发生与小胶质细胞极化密切相关，ATP敏感性钾通道（KATP）参与了小胶质细胞极化。H2S是神经系统重要的信号分子，调控细胞内外信息通讯，发挥神经保护作用。申请人将自主设计的新型H2S供体用于癫痫模型发现：①给予低浓度H2S可抑制大鼠癫痫发作并上调KATP蛋白表达（Zhu X et al. APPL MATER TODAY, 2020）；②小胶质细胞P2Y12受体敲除可加剧小鼠癫痫发作；③预实验显示：H2S可上调KATP蛋白表达，敲除小鼠P2Y12则降低其表达。然而，H2S调控P2Y12/KATP的具体机制及其与癫痫的关系尚未明确。本项目拟建立细胞及动物模型，利用基因修饰、电生理等手段，探讨H2S调控P2Y12和KATP蛋白表达及其与小胶质细胞极化的关系，分析KATP电流与痫性放电的规律，揭示H2S调控P2Y12/KATP功能促进小胶质细胞极化抑制癫痫的分子机制，为癫痫防治提供新思路。

The pathological process of epilepsy is closely related to microglia polarization in which ATP sensitive potassium channel (KATP) has been proved to be involved. Hydrogen sulfide (H2S), an important signaling molecule of the nervous system, regulates the intracellular and extracellular communications, and also plays a role in neural protection. We synthesized a novel H2S donor for animal models of epilepsy and have several findings. Firstly, the exogenously administered H2S at low concentration suppressed seizure in a rat model and up-regulated the protein expression of KATP in neurons(Zhu X et al. Applied Materials Today, 2020). Secondly, knockout of P2Y12 receptor in microglia led to worsening of seizure in mice. Furthermore, our preliminary results showed that H2S increased the expression of KATP protein in microglia, while decreased the protein expression in the knockout of P2Y12 mice. However, the molecular mechanism of H2S regulating P2Y12 receptor and KATP to inhibit epilepsy remains unclear. Hence, we establish epileptic models in vitro and in vivo and use different kinds of experimental methods including genetic modification and electrophysiology to clarify this underlying mechanism. The project has two main aims: to explore how H2S modulates the function of P2Y12 receptor and KATP protein as well as the interaction with microglial polarization; and to illustrate the relationship between KATP current in microglia and neuronal epileptic discharges. These results may provide promising ways for epilepsy treatment.