钙调素抑制剂DY98抑制NLRP3炎症小体活化和细胞焦亡改善血管性痴呆认知功能的研究

Vascular dementia (VD) is a chronic progressive disease with cognitive impairment induced by cerebrovascular disease and cerebral blood flow hypoperfusion. An abnormal increase in calmodulin (CaM) activity in calcium overload may activate CaM-dependent multiple pathways. This is associated with VD-induced cognitive impairment, but whether calmodulin induce activation of NLRP3 inflammasome on downstream causes cell pyroptosis is unknown. Our previous study demonstrated that DY98, CaM inhibitor, could inhibit of CaM may improve cognitive impairment of VD. The aim of the study is to validate the effect of DY98 is self-designed and self-prepared on cognitive impairment induced by bilateral carotid artery occlusion of VD in mouse, to evaluate the behavioral changes of cognitive function associated Y maze, new object recognition and inhibitory avoidance tasks, as well as explore the mechanism of DY98 to alleviate neuronal and white matter damage, focusing on nitrification stress and inflammatory NLRP3 activation induced inflammation changes after inhibition of CaM. New drugs is expected for treatment VD will be discovered.

血管性痴呆（VD）是由卒中和大脑低灌注等血管性疾病引起认知损伤的综合征。钙超载时，钙调素（CaM）活性异常增高，激活靶酶，可否活化下游的NLRP3炎症小体致细胞焦亡（pyroptosis），引起VD认知障碍尚属未知。我们前期报道CaM抑制剂DY98可抑制CaM，改善VD认知功能损伤。本研究采用经典双侧颈总动脉狭窄法制备血管性痴呆小鼠模型，应用自主设计和制备的DY98，使用旷场实验、新物体识别、Y迷宫和被动回避的行为学方法，评价DY98改善认知的药效学作用，探索DY98缓解神经元和脑白质损伤的细胞焦亡机制，重点关注抑制CaM后硝化应激和炎症小体NLRP3激活诱导炎症和焦亡的改变。可望寻找作用于炎性通路上游的药物，有望在疾病的早期阶段预防或延缓VD疾病进程。

血管性痴呆（VD）是由卒中和大脑低灌注等血管性疾病引起认知损伤的综合征。钙超载时，钙调素（CaM）活性异常增高，激活靶酶，可否活化下游的NLRP3炎症小体致细胞焦亡（pyroptosis），引起VD认知障碍尚属未知。因此，本研究旨在探索自主设计和制备的CaM抑制剂DY98可抑制CaM，改善VD认知功能损伤。本项目采用右侧颈总动脉结扎法制备血管性痴呆小鼠模型，口服给予DY98，行为学实验证实其可提高空间和非空间学习记忆能力。小鼠脑组织病理切片H.E染色观察到DY98可以改善海马区和胼胝体损伤；免疫荧光双染证实DY98可通过抑制小胶质细胞活化，降低白细胞介素-1（IL-1）水平，且抑制NLRP3/Caspase-1/GSDMD信号通路激活对血管性痴呆小鼠的认知功能障碍有改善作用。后续在体外实验证实，DY98不仅可减轻LPS+ATP诱导的N9小胶质细胞细胞和星形胶质细胞NLRP3炎性激活和细胞焦亡，且可减轻TNF-α诱导的少突胶质细胞焦亡，其机制均与抑制NLRP3引起的经典焦亡通路相关。综上所述，DY98抑制神经胶质细胞发生细胞焦亡，其在 NLRP3 活化和细胞焦亡通路上游起阻断作用，抑制VD炎性病理进程，进而改善认知障碍。本研究为探索VD疾病的早期阶段预防或延缓疾病进程，提供新方向和新策略。