RIP1/RIP3/MLKL介导“程序性坏死”在银屑病中的作用与调控机制研究

银屑病皮损区角质形成细胞死亡调节异常，申请者预实验发现，一种新型细胞死亡方式──“程序性坏死”参与了银屑病炎症的发生，并且与自噬“串扰（Crosstalk）”共同决定角质形成细胞的命运，其与炎症的关系有待阐明。RIPK1-RIPK3-MLKL是启动TNF-α诱导程序性坏死的经典途径，但调控机制尚需进一步研究，与其它细胞死亡方式如自噬的“串扰”尚不清楚，在银屑病中更未见报道。本课题拟在既往研究基础上，进行线粒体膜电位的检测、细胞死亡形态的电镜观测，采用RNA测序、特异性抑制剂、基因敲除和过表达等手段，从细胞、动物、患者的水平，深入研究RIPK1-RIPK3-MLKL信号途径在银屑病中“促炎”机制、RIPK1/RIPK3-ROS信号途径对程序性坏死、自噬的影响、MK2调控RIPK1磷酸化的作用等，以明确程序性坏死促炎的关键靶点，探讨协同调控程序性坏死和自噬的可行性，为银屑病的治疗提供新思路。

The regulation of keratinocyte cell death in psoriatic lesions is abnormal. Applicant's preliminary experiment found that a new way of cell death ----"programmed necrosis" is involved in the occurrence of psoriasis inflammation. It can crosstalk with autophagy to jointly determine the fate of keratinocytes. However, its relation to inflammation remains to be elucidated. RIPK1-RIPK3-MLKL is a classical pathway to initiate TNF-α-induced programmed necrosis, but the regulatory mechanism needs further study. The effect of “crosstalk” with other cell death modes, such as autophagy, remains unclear, and there is a lack of such report in psoriasis. Based on previous studies, this project will detect mitochondrial membrane potential and electron microscopic observation of cell death morphology and will adopt RNA-seq, specific inhibitors, and gene knockout and overexpression to systematically investigate the "proinflammation" mechanism of RIPK1-RIPK3-MLKL signaling pathway in psoriasis, the RIPK1/RIPK3-ROS signaling pathway in programmed necrosis and autophagy, and the phosphorylation of RIPK1 by MK2 from the level of cells, animals and patients. Not only is this study expected to determine the key target of proinflammation of programmed necrosis and discuss the feasibility of synergistic regulation of programmed necrosis and autophagy, but also to provide new perspective for the treatment of psoriasis.