基于SCN-海马神经环路探讨昼夜节律紊乱促进颞叶癫痫发作的分子机制及“从肝论治”的干预作用

颞叶癫痫多在白昼发作，昼夜节律紊乱明显影响癫痫的发作与预后。海马的“生物钟蛋白”BMAL1/PER的改变可致神经元异常去极化，促发癫痫，但昼夜节律起搏器视交叉上核（SCN）如何影响颞叶癫痫尚不清楚。为此提出假设：昼夜节律紊乱时，Kv4.2介导的IA引起SCN兴奋性改变，通过BMAL1/PER等调节SCN-海马神经环路，促进颞叶癫痫的发作。前期研究颞叶癫痫海马神经元Kv4.2表达减少，IA幅度降低，“从肝论治”方药改善癫痫后Kv4.2及IA抑制。为验证假说，利用BMAL1条件性敲除及过表达小鼠，光照调节模拟昼夜节律紊乱，建立颞叶癫痫及脑片模型，用“从肝论治”方药及Kv4.2工具药干预，AAV荧光示踪标记SCN-海马神经环路，膜片钳检测IA及在体多通道检测SCN神经元信号等，探讨昼夜节律紊乱时，SCN-海马神经环路影响癫痫的重要机制，以及“从肝论治”的干预作用，为治疗提供新的药物靶点。

Seizures of temporal lobe epilepsy mostly occur in the daytime, and the circadian rhythm disorder significantly affects the onset and prognosis of epilepsy. The changes of circadian protein BMAL1 /PER in the hippocampus can cause abnormal depolarization of neurons, inducing the temporal lobe epilepsy. But it is unclear how the circadian pacemaker Suprachiasmatic Nucleus (SCN) affects temporal lobe epilepsy. We propose the hypothesis that circadian rhythm disorder will cause Kv4.2-mediated IA to suppress the excitability of SCN, and regulate the SCN-hippocampal neural circuit through protein expression changes such as BMAL1 /PER, resulting in temporal lobe epilepsy. In our previous study, the decrease of Kv4.2 in hippocampal neurons of temporal lobe epilepsy reduced IA amplitude, which was improved by the prescription of "liver associated treatments ".To verify the hypothesis, we will establish the temporal lobe epilepsy model and brain slice model using BMAL1 conditional knockout and overexpress mice, simulate circadian rhythm disorder with light adjustment, use the prescription of "liver associated treatments" and Kv4.2 channel opener/blocker to intervene, label the SCN-hippocampal neural circuit with AAV fluorescence tracing technique, record IA current with patch-clamp technique and detect SCN neuronal signals with multi-channel in-vivo recording technology. We intend to explore the important mechanism of the SCN-hippocampal neural circuit affecting epilepsy and the effect of " liver associated treatments" under the disorder of circadian rhythm. This work will provide potential targets for the treatment of epilepsy.