IL-17A激活NLRP3介导细胞焦亡在氟中毒动物睾丸组织细胞损伤中的调节机制

细胞焦亡作为新近发现的一种程序性死亡方式，在各种疾病与组织损伤中起重要调节作用，而对是否参与动物氟中毒组织损伤调节未见报道。本项目利用已建立的IL-17A基因敲除与功能增强氟中毒小鼠模型，采用电镜分析、酶联免疫、免疫荧光、定量PCR和蛋白印迹等技术研究睾丸组织细胞形态、焦亡关键蛋白GSDMD、NLRP3、Caspase-1、ASC、Pro-IL-1β、IL-1β、IL-18等表达；并进一步通过体内外实验，上调及阻断NLRP3炎症体通路，研究上述焦亡关键蛋白和基因，IL-17A和受体、IL-17A 分泌细胞因子以及调节通路中NF-κB、IFN-γ、TNF-α等基因和蛋白质表达，探讨NLRP3介导的细胞焦亡与IL-17A通路之间的关系，解析IL-17A激活NLRP3炎症小体介导细胞焦亡在氟致睾丸组织细胞损伤的机制，旨在阐明氟的雄性生殖毒性机理，为从分子水平上开展氟中毒的诊断和治疗提供新的靶点。

Pyrocytosis, as a newly discovered type of programmed cell death, plays an important regulatory role in various diseases and tissue damages, and whether it is involved in the regulation of tissue damage caused by animal fluorosis has not been reported. This project will study the testicular tissue cell morphology, pyrolysis key protein GSDMD, NLRP3, Caspase-1, ASC, Pro-IL-1β, IL-1β, IL-18 and other expressions by using various techniques including electron microscopy, ELISA, immunofluorescence, quantitative PCR and Western blotting based on the established mice models of IL-17A gene knockout and functional enhancement treated with fluoride. Furthermore, other models of in vivo and in vitro experiments, and up-regulate and block the NLRP3 inflammatory pathway will be established to study the above key proteins and genes of pyrolysis, IL -17A and receptors, IL-17A secrete cytokines, and regulate the expression of genes and proteins such as NF-κB, IFN-γ, and TNF-α in the IL-17A pathway, and explore the relationship between NLRP3-mediated cell pyrolysis and IL-17A pathway. The objective of all analyze is to explore the mechanism of IL-17A activating NLRP3 inflammasome mediated pyrocytosis in testicular tissue damage caused by fluoride, which aims to elucidate the mechanism of male reproductive toxicity of fluorine, and provide a new target for the diagnosis and treatment of fluorine poisoning at the molecular level.