S1P调控NLRC4介导的星形胶质细胞焦亡在ASD认知和社交损伤中的作用及机制研究

孤独症谱系障碍(ASD)是一种严重的神经发育障碍性疾病，核心症状为社交障碍，多伴有认知损伤。前期研究提示焦亡相关的异常神经炎性反应可能与ASD相关，而1-磷酸鞘氨醇(S1P)可能参与调控该作用。本研究以ASD儿童及孤独症模型鼠为研究对象，阐明异常的神经炎性反应与ASD的关联，并通过鞘氨醇激酶阻断干预明确S1P与ASD认知及社交损伤的关联及对星形胶质细胞焦亡的调控作用；同时，构建NLRC4基因敲除鼠探讨S1P是否通过调控NLRC4介导星形胶质细胞焦亡并造成海马神经元形态及功能损伤。此外，分别构建NLRC4、caspase-1、GSDMD基因敲除的SMA-560细胞株并通过基因回补方式，观察S1P干预后焦亡相关蛋白表达及炎性因子水平的变化，从细胞和分子水平共同阐明S1P调控NLRC4介导星形胶质细胞焦亡的作用及机制，证实ASD认知及社交损伤的神经炎症假说，为ASD的靶向干预提供新的思路及方向。

Autism spectrum disorder (ASD) is a heterogeneous group of neurodevelopmental disorder, which is characterized as impairment in social function and most accompanied by cognitive deficits. Our prior work has reported that abnormal neuroinflammatory response derived from pyroptosis may be associated with ASD, and this process could be regulated by S1P. In the current study, the autistic children and BTBR mice (ASD model mice) are used as the research objects to analyze the relationship between abnormal neuroinflammatory response and ASD. Furthermore, we investigate the effect of S1P on the cognitive and social functioning of ASD model mice and on the pyroptosis of astrocytes via treating with inhibitors of S1P synthetase (SKI-II). NLRC4 knockout mice (based on BTBR mice) are constructed to explore whether S1P is involved in NLRC4 inflammasome-mediated pyroptosis of astrocytes, subsequently damaging hippocampal neuron morphology and function. In addition, we build SMA-560 cell lines with knockout of NLRC4, caspase-1 and GSDMD gene, respectively and then supplement these genes, to observe the changes of pyroptosis-related proteins and inflammatory factor after S1P intervention. From the cellular and molecular levels, we attempt to demonstrate the effect and mechanism that S1P regulates NLRC4 inflammasome-mediated pyroptosis of astrocytes, to consolidate the hypothesis that ASD cognitive and social impairment is caused by abnormal neuroinflammatory response, which could provide new insight for targeted intervention of ASD.