姜黄素调节NLRP1炎症小体活化介导的神经元焦亡治疗脑缺血再灌注损伤作用机制研究

Neuronal death induced by Ischemia/Reperfusion (I/R) is the main reason of ischemic brain injury. Neuronal pyroptosis is involved in the development of neurodegenerative diseases. Our previous study showed that curcumin has a therapeutic effect on cerebral ischemia-reperfusion injury, and curcumin down-regulated the expression of NLRP1 and Caspase-1,which are the key molecules of Pyroptosis, and played a protective role on neurons. This project intends to establish a rat model of middle cerebral artery occlusion (MCAO) and neuronal cell models of oxygen-glucose deprivation (OGD) in vitro to clarify the role and the mechanism of neuronal pyroptosis on neuronal death in cerebral ischemia-reperfusion injury through silencing or over-expression of NLRP1 and Caspase-1 mediated by lentiviral, electron microscopy scanning nerve cell structure, detecting the expression of NLRP1, Caspase-1 and GSDMD by immunofluorescence and immunoblotting and the expression of IL-1β by ELISA. Fuethermore, the role of TLR4/MAPK/NF-κB signaling on neuronal pyroptosis will be analyzed and the molecular mechanism of curcumin inhibition of neuronal pyroptosis will be elucidated. The project will further elucidate the mechanism of cerebral ischemia-reperfusion injury and provide a guidance for the prevention and treatment and the related drugs development of cerebral ischemia-reperfusion injury.

缺血再灌注(I/R)导致神经元死亡是缺血性脑损伤主要原因。神经细胞焦亡参与神经退行性疾病的发生；我们前期研究姜黄素对脑I/R有治疗作用，并发现姜黄素下调细胞焦亡关键分子NLRP1和Caspase-1的表达，提示姜黄素通过神经元细胞焦亡发挥对神经元的保护作用。本项目拟建立大鼠大脑中动脉缺血再灌注(MCAO)模型及神经元体外氧糖剥离(OGD)细胞模型，通过慢病毒沉默或过表达NLRP1和Caspase-1，电镜扫描观察神经细胞结构，免疫荧光、免疫印迹法检测NLRP1、Caspase-1、GSDMD的表达，ELISA检测IL-1β表达水平，阐明细胞焦亡在脑I/R损伤中神经元死亡中的作用及机制。并分析TLR4/MAPK/NF-κB信号在调控神经细胞焦亡中的作用，阐明姜黄素抑制神经细胞焦亡的分子机制。本项目将进一步阐明I/R诱导脑损伤的机制，对脑I/R的防治及相关药物的研发具有指导意义。

缺血再灌注(I/R)导致神经元死亡是缺血性脑损伤主要原因。神经细胞焦亡参与神经退行性疾病的发生；我们前期研究姜黄素对脑I/R有治疗作用，并发现姜黄素下调细胞焦亡关键分子NLRP1和Caspase-1的表达，提示姜黄素通过神经元细胞焦亡发挥对神经元的保护作用。本项目拟建立大鼠大脑中动脉缺血再灌注(MCAO)模型及神经元体外氧糖剥离(OGD)细胞模型，通过慢病毒沉默或过表达NLRP1和Caspase-1，电镜扫描观察神经细胞结构，免疫荧光、免疫印迹法检测NLRP1、Caspase-1、GSDMD的表达，ELISA检测IL-1β表达水平，阐明细胞焦亡在脑I/R损伤中神经元死亡中的作用及机制。并分析TLR4/MAPK/NF-κB信号在调控神经细胞焦亡中的作用，阐明姜黄素抑制神经细胞焦亡的分子机制。本项目将进一步阐明I/R诱导脑损伤的机制，对脑I/R的防治及相关药物的研发具有指导意义。