■NOD样受体家族含pyrin结构域3(NLRP3)炎性体,通过调节IL-1β在感染性和炎症性疾病中至关重要,提出了疾病管理的目标。淋病奈瑟菌每年导致超过8700万人淋病,与先前的研究揭示NLRP3炎性体在感染的巨噬细胞中激活。没有报道天然产物抵消这种活化。探索和厚朴酚,一种来自中草药的酚类化合物,我们研究了其对淋病奈瑟菌感染的巨噬细胞中NLRP3炎性体活化的影响。使用ELISA和Western印迹分析
■和厚朴酚对促炎介质的蛋白表达的影响。通过特异性荧光探针(CM-H2DCFDA和MitoSOX,分别)并通过流式细胞术进行分析。使用特异性荧光探针(MitoTracker和DiOC2(3))评估线粒体膜完整性并通过流式细胞术分析。此外,和厚朴酚对淋病奈瑟菌活力的影响通过体外集落形成单位测定来检测。
■和厚朴酚有效抑制caspase-1,caspase-11和GSDMD的激活,并减少IL-1β的细胞外释放,淋病奈瑟菌感染的巨噬细胞中NLRP3和含有半胱天冬酶募集结构域(ASC)的凋亡相关斑点样蛋白。详细的研究表明,和厚朴酚可降低淋病奈瑟菌感染的巨噬细胞中H2O2的产生和ERK1/2的磷酸化。重要的是,JNK1/2和p38的磷酸化和NF-κB的激活不受影响。此外,和厚朴酚减少了淋病奈瑟菌介导的线粒体内活性氧的产生,保持他们的完整性。此外,和厚朴酚抑制淋病奈瑟菌诱导的促炎介质IL-6和诱导型一氧化氮合酶的表达,而与NLRP3无关。令人印象深刻的是,和厚朴酚对淋病奈瑟菌具有体外抗淋球菌活性。
■和厚朴酚抑制淋病奈瑟菌感染的巨噬细胞中的NLRP3炎性体,并有望进一步发展为预防和治疗淋病相关症状的活性成分。
UNASSIGNED: The NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome, crucial in infectious and inflammatory diseases by regulating IL-1β, presents a target for disease management. Neisseria gonorrhoeae causes gonorrhea in over 87 million people annually, with previous research revealing NLRP3 inflammasome activation in infected macrophages. No natural products have been reported to counteract this activation. Exploring
honokiol, a phenolic compound from Chinese herbal medicine, we investigated its impact on NLRP3 inflammasome activation in N. gonorrhoeae-infected macrophages.
UNASSIGNED: Honokiol\'s impact on the protein expression of pro-inflammatory mediators was analyzed using ELISA and Western blotting. The generation of intracellular H2O2 and mitochondrial reactive oxygen species (ROS) was detected through specific fluorescent probes (CM-H2DCFDA and MitoSOX, respectively) and analyzed by flow cytometry. Mitochondrial membrane integrity was assessed using specific fluorescent probes (MitoTracker and DiOC2(3)) and analyzed by flow cytometry. Additionally, the effect of
honokiol on the viability of N. gonorrhoeae was examined through an in vitro colony-forming units assay.
UNASSIGNED: Honokiol effectively inhibits caspase-1, caspase-11 and GSDMD activation and reduces the extracellular release of IL-1β, NLRP3, and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) in N. gonorrhoeae-infected macrophages. Detailed investigations have demonstrated that honokiol lowers the production of H2O2 and the phosphorylation of ERK1/2 in N. gonorrhoeae-infected macrophages. Importantly, the phosphorylation of JNK1/2 and p38 and the activation of NF-κB remain unaffected. Moreover, honokiol reduces the N. gonorrhoeae-mediated generation of reactive oxygen species within the mitochondria, preserving their integrity. Additionally,
honokiol suppresses the expression of the pro-inflammatory mediator IL-6 and inducible nitric oxide synthase induced by N. gonorrhoeae independently of NLRP3. Impressively,
honokiol exhibits in vitro anti-gonococcal activity against N. gonorrhoeae.
UNASSIGNED: Honokiol inhibits the NLRP3 inflammasome in N. gonorrhoeae-infected macrophages and holds great promise for further development as an active ingredient in the prevention and treatment of symptoms associated with gonorrhea.