JAM4在癫痫持续状态形成中的分子机制

The epileptic state is an important cause of death in the neurologic intensive care unit (ICU). Its occurrence is related to the weakening or disappearance of the refractory period after epileptic attack. Due to the phenomenon of epileptic tolerance, there are different responses to the different neurotransmitters, and it can significantly increase the efficacy of antiepileptic drugs. Therefore，we identified that there is a specific neurochemical mechanism has taken part in its formation.Using the maximum electric shock, acoustic epileptic rat and the model of meriones unguiculatus with epileptic susceptibility to combine with the large flux gene and protein chip scanning, the group found that JAM4 plays an important role in the weakening or disappearance of the epilepsy refractory period. After we obtain the expected results through embryo transfer, patch clamp, exocrine scan. This topic use that the research group has established the brain trust of more than 300 cases of brain tissue of epileptic patients after operation，the clinical resource bank of more than 1.5 million epileptic patients for regular follow-up，and the genetic resource base of more than 600 epilepsy hereditary family based on the previous research. We continue to explore the molecular mechanisms of JAM4 in the formation of the epileptic state to find the new ideas and new methods for the prevention and treatment of the epileptic state based on the model of the lentivirus intervention.

癫痫持续状态是神经重症病房中患者死亡的重要原因，其发生与癫痫发作后不应期的减弱或消失有关。由于这种癫痫耐受现象对不同神经递质有不同的反应，且能明显增加抗癫痫药物的疗效，因而认定有特定的神经化学机制参与了其形成。利用最大电休克、声源性癫痫鼠及癫痫易感性长爪沙鼠模型结合大通量基因和蛋白芯片扫描，课题组发现JAM4在癫痫不应期减弱或消失中起着重要作用。通过胚胎电转、膜片钳、外泌体扫描取得预期结果后，本课题拟在前期研究基础上，利用课题组建立的有300多例癫痫患者手术后脑组织组成的脑库和由1.5万多癫痫患者定期随访的临床资源库及有600多个癫痫遗传家系组成的遗传资源库，以慢病毒干预模型为基础，继续探索JAM4在癫痫持续状态形成中的分子机制，寻找癫痫持续状态防治的新思想和新方法。

我们首先用化学遗传技术对癫痫持续状态发作后受影响最明显的26个核团进行了研究，结果发现只有在海马CA区和后脑同步刺激谷氨酸能神经元才能诱导出动物的癫痫持续状态，用多导电生理记录仪同步进行的的记录也发现其点燃动物的场电位与人类的癫痫持续状态非常相似，从而建立起一种内源性癫痫持续状态模型。. 为了解JAM4与癫痫间的关系，首先我们收集了20例曾有癫痫持续状态发作患者手术后的脑组织标本，通过免疫荧光法和蛋白印记法检测了其与对照组脑组织中JAM4的表达。结果发现JAM4与神经元标志物MAP2共表达与神经胶质细胞标志物GFAP无共表达，且癫痫患者脑组织中JAM4表达量明显高于对照组；同时我们在戊四氮（PTZ）和海人酸（KA）诱导癫痫持续状态的模型中也发现JAM4的表达在神经细胞上，而胶质细胞上无表达，这一结果与人脑组织的研究一致，用蛋白印迹的方法我们还发现在KA小鼠皮质和海马区中也发现JAM4在癫痫组有更高的表达。为证明JAM4表达量的改变是否对癫痫行为产生影响，我们通过敲低或过表达JAM4，结果发现JAM4-敲低小鼠癫痫发作的强度与对照组低，而JAM4-过表达小鼠则出现较强的发作级别，另一方面，JAM4过表达产生较高SRSs频率。同时我们还对海人酸癫痫小鼠进行脑电监测，记录并对比其30分钟内海马区局部场电位（LFPs）的改变，结果发现敲低组癫痫样放电次数（SLEs）减少，过表达组SLEs增多，这一结果与行为学改变一致。因此，我们认为JAM4在癫痫发作后不应期消失中起着重要作用，这些发现有助于更好的了解癫痫持续状态形成的机制。