小肠结肠炎综合征小鼠肠黏膜中NLRP3表达与炎症分子关系的研究

Relationship between NLRP3 and inflammatory molecules expression in intestinal mucosa cells of mice with FPIES

目的探讨食物蛋白诱导的小肠结肠炎综合征(FPIES)小鼠肠黏膜细胞中炎症小体NOD样受体热蛋白结构域相关蛋白3(NLRP3)表达水平,明确NLRP3异常与炎症分子及细胞焦亡的关联性。方法利用卵清蛋白灌胃建立食物蛋白诱导FPIES小鼠模型;实时荧光定量聚合酶链反应(qPCR)和Western blot方法检测肠黏膜细胞NLRP3、炎症分子及细胞焦亡通路分子的表达水平;采用药物干预黏膜细胞,分别抑制和激活NLRP3,Western blot方法检测肠黏膜细胞炎症分子及细胞焦亡通路分子表达的改变情况。结果FPIES小鼠肠黏膜细胞中NLRP3、炎症分子及细胞焦亡通路分子表达水平显著上调(P<0.05);激活NLRP3表达,可以诱导炎症分子及细胞焦亡通路分子表达显著上调,抑制NLRP3表达,可以显著上调炎症分子转化生长因子(TGF)-β和肿瘤坏死因子(TNF)-α表达(P<0.05)。结论FPIES小鼠肠黏膜细胞NLRP3表达与炎症反应及细胞焦亡密切相关,是调控FPIES肠道病理表型的上游靶标分子。

Objective To investigate the expression level of NLRP3 in intestinal mucosal cells of mouse model with food protein-induced enterocolitis syndrome(FPIES),and to clarify the relationship between inflammatory molecules and cell pyroptosis induced by abnormal expression of NLRP3.Methods The FPIES mouse model was established by ovalbumin intragastric administration.Fluorescent quantitative PCR and Western blot methods were used to detect the expression levels of NLRP3,inflammatory molecules and pyrolytic pathway molecules.Western blot assay was used to detect the expression levels of inflammatory molecules and pyrolytic pathway molecules in intestinal mucosal cells after activating or inhibiting NLRP3.Results The expression levels of NLRP3,inflammatory molecules and pyrolytic pathway molecules in intestinal mucosal cells were significantly up-regulated in FPIES model mice(P<0.05).The activation of NLRP3 expression could significantly up-regulate the expression of inflammatory molecules and pyrolytic pathway molecules,and the inhibition of NLRP3 expression could significantly up-regulate the expression of TGFβand TNFα(P<0.05).Conclusion The expression of NLRP3 in intestinal mucosal cells is closely related to the inflammatory response and cell pyroptosis in FPIES model mice,which is the upstream target molecule to regulate FPIES intestinal pathological phenotypes.