贝毒azaspiracid-1诱导肾细胞内质网应激-自噬相互调节的分子机制研究

Azaspiracid(AZA) is a new kind of shellfish toxin, which is responsible for a series of human intoxication. AZA-1 is one representative type of the AZA family. The previous studies show that AZA-1 has toxicity to many cell lines from important organs of human. So it is necessary to study the toxicity mechanism. Our early study found that AZA-1 induced autophagy and endoplasmic reticulum stress(ER stress) which could regulate each other in kidney cells. In view of the important biological functions of autophagy and ER stress, we consider that the mutual regulation phenomenon may be related to the renal toxicity induced by AZA-1. The concentration of intracellular free Ca2+ induced by AZA-1 and the expression of Ca2+-dependent autophagy-related protein will be detected and the relationship between Ca2+ and ER will be measured to study the signaling pathway that ER stress activates autophagy mediated by Ca2+. Moreover, the expression level of p62 will be measured and the hypothesis that autophagy activates ER stress mediated by p62 will be verified, and then its signaling pathways will be explored by considering autophagy flux. Based on the above researches, the mutual regulation mechanism between autophagy and ER stress induced by AZA-1 will be elucidated finally which will provide new theoretical basis for AZA-1 prevention and control.

原多甲藻酸(azaspiracid, AZA)是一类新型贝类毒素，因食用AZA污染的贝类引发的中毒事件屡有报道，AZA-1是AZA家族中最具代表性的一种，国内外研究表明AZA-1对人体重要器官的细胞系均具有毒性，因此研究其毒性产生的机制非常必要。前期实验发现AZA-1诱导肾细胞产生自噬和内质网应激且二者可以相互调控，鉴于自噬和内质网应激具有重要的功能，因此二者相互调控的现象与AZA-1诱导的肾毒性有较大关联。本研究拟通过测定AZA-1诱导的胞内游离Ca2+的浓度，检测依赖Ca2+的自噬相关蛋白的表达及分析Ca2+与内质网的关系，研究内质网应激通过Ca2+激活自噬的信号通路；同时检测AZA-1诱导的p62蛋白的水平并验证p62介导自噬调控内质网应激的猜想，结合自噬流量监测的结果深入探索其调控通路；最终阐明AZA-1诱导的自噬和内质网应激相互调控的机制，为制定危害防治措施提供新的理论依据。

AZA1是一种原多甲藻酸贝类毒素, 误食AZA1严重威胁到了人和哺乳动物的健康, 并且对肝肾的损伤尤为严重。研究发现，AZA1可以诱导自噬和内质网应激, 并且这两者存在相互调控作用。通过RNA-seq，全蛋白质谱技术，高通量药物筛选技术，蛋白质相互关系等研究发现p62在自噬和内质网应激调控的细胞肝肾损伤过程中起关键作用，p62-IRE1α-XBP1通路是参与自噬和内质网应激调控的细胞增殖的主要信号通路。阐明AZA1诱导肝肾细胞损伤的机制，为筛选和开发保肝解毒的功能食品提供了基础。