沉默MLKL基因抑制坏死性凋亡对幼年大鼠癫痫持续状态后的脑保护作用研究

Epilepsy is a common neurological disease in childhood, and status epilepticus (SE) is one of the most critical conditions in pediatric emergency. Neuron death caused by SE is the pathological basis of the occurrence of chronic spontaneous seizures. It has great significance to clarify the mechanism of neuron death in the process of searching novel and effective methods to protect neurons and decrease the development of refractory epilepsy. Necroptosis is a newly recognized programmed cell death, and our preliminary studies demonstrate that necroptosis is probably engaged in the neuron death of amygdale and piriform cortex of juvenile rats after SE. Therefore, we consider the possibility to decrease injury to the developing brain after SE by blocking necroptosis. Mixed lineage kinase domain-like (MLKL) protein plays critical roles in necroptosis and is the executor of cell death. To provide new strategies for the treatment of childhood epilepsy, in this study, we will knock out MLKL gene with siRNA, and investigate whether inhibiting necroptosis can reduce injury to immature brain, improve cognition and block the process of epileptogenesis after SE. Furthermore, in view of the significant roles of inflammatory factors in epileptogenesis, we will perform an initial evaluation for the relationship of necroptosis and cytokines with epileptogenesis, and provide theoretical basis for applying anti-inflammatory therapy in modulating epileptogenesis.

癫痫是儿童神经系统常见疾病，而儿童期的癫痫持续状态（status epileticus，SE）是儿科的危急重症。SE导致的神经元死亡是慢性自发性癫痫发生的病理基础。明确SE后神经元死亡的机制对于寻求新的、有效的癫痫后神经元保护方法，减少难治性癫痫的发生，具有重要的意义。坏死性凋亡是新近发现的一种程序性细胞死亡，我们的前期研究发现，坏死性凋亡与幼鼠SE后杏仁核和梨状皮质的神经细胞死亡机制密切相关。因此提出设想：通过抑制坏死性凋亡减轻SE对幼鼠发育期大脑的损害。MLKL在坏死性凋亡中发挥关键作用，是死亡的执行蛋白。本研究拟通过 SiRNA敲除MLKL基因，抑制坏死性凋亡，明确抑制坏死性凋亡对幼鼠SE后的脑保护作用、改善认知功能以及减少慢性期癫痫发生的作用。同时，鉴于炎症因子在癫痫发生中的重要作用，将初步探讨坏死性凋亡、炎症因子与癫痫发生的关系，为临床治疗儿童期癫痫提供新思路。

癫痫是儿童神经系统常见疾病，而儿童期的癫痫持续状态（status epileticus，SE）是儿科的危急重症。SE导致的神经元死亡是慢性自发性癫痫发生的病理基础。明确SE后神经元死亡的机制对于寻求新的、有效的癫痫后神经元保护方法，减少难治性癫痫的发生，具有重要的意义。本研究发现，幼鼠SE后主要的脑损伤部位在海马、杏仁核和梨状皮质。细胞焦亡标志物gasdermin D的表达增加，而铁死亡的标志物GPX4表达降低，表明细胞焦亡和铁死亡这两种新型的死亡方式参与了幼鼠SE后的脑损伤。gasdermin D的表达增加先于GPX4的表达降低。进一步的研究将探讨Gasdermin D通过铁自噬诱导SE后神经元铁死亡的作用机制，并通过体内敲除Gasdermin D，研究干预后对幼鼠大脑的保护作用、改善认知功能以及减少慢性期癫痫发生的作用，为临床治疗儿童期癫痫提供新思路。