氟化物对免疫系统的损害是全世界关注的公共卫生问题,然而,潜在的机制尚不清楚。核黄素和IL-17A均与免疫功能密切相关,并调节氟化物的睾丸毒性。然而,核黄素或IL-17A是否参与氟化物诱导的免疫毒性尚不清楚.这里,我们首先通过饮用水用氟化钠(NaF)(100mg/L)处理小鼠91天,建立了雄性ICR小鼠模型。结果表明,氟增加了促炎因子IL-1β和IL-17A的表达,导致脾脏炎症和形态学损伤。此外,核黄素转运蛋白SLC52A2和SLC5A3的表达水平;转化相关酶RFK和FLAD1;以及关键的线粒体功能决定子SDH,COX,通过实时PCR检测脾脏中的ATP,西方印迹,和ELISA。结果显示氟化物破坏了核黄素的转运,改造,新陈代谢,和线粒体功能。此外,野生型(WT)和IL-17A敲除(IL-17A-/-)C57BL/6J相同年龄的雄性小鼠用NaF(24mg/kg·bw,相当于100mg/L)和/或核黄素磷酸钠(5mg/kg·bw)通过管饲法进行91天。如上所述评估类似的参数。结果证实氟化物通过RFK而不是通过FLAD1增加核黄素代谢。氟化物还影响脾脏中的线粒体功能和活化的嗜中性粒细胞(用Ly6g标记)和巨噬细胞(用CD68标记)。有趣的是,IL-17A部分介导氟化物诱导的脾脏核黄素代谢紊乱和免疫毒性。这项工作不仅揭示了氟化物的新毒性机制,而且为探索核黄素的生理功能以及诊断和治疗氟化物在环境中的毒性作用提供了新的线索。
The impairment of the immune system by
fluoride is a public health concern worldwide, yet the underlying mechanism is unclear. Both riboflavin and IL-17A are closely related to immune function and regulate the testicular toxicity of
fluoride. However, whether riboflavin or IL-17A is involved in
fluoride-induced immunotoxicity is unknown. Here, we first established a male ICR mouse model by treating mice with sodium
fluoride (NaF) (100 mg/L) via the drinking water for 91 days. The results showed that
fluoride increased the expression of the proinflammatory factors IL-1β and IL-17A, which led to splenic inflammation and morphological injury. Moreover, the expression levels of the riboflavin transporters SLC52A2 and SLC52A3; the transformation-related enzymes RFK and FLAD1; and the key mitochondrial functional determinants SDH, COX, and ATP in the spleen were measured via real-time PCR, Western blotting, and ELISA. The results revealed that fluoride disrupted riboflavin transport, transformation, metabolism, and mitochondrial function. Furthermore, wild-type (WT) and IL-17A knockout (IL-17A-/-) C57BL/6 J male mice of the same age were treated with NaF (24 mg/kg·bw, equivalent to 100 mg/L) and/or riboflavin sodium phosphate (5 mg/kg·bw) via gavage for 91 days. Similar parameters were evaluated as above. The results confirmed that fluoride increased riboflavin metabolism through RFK but not through FLAD1. Fluoride also affected mitochondrial function and activated neutrophils (marked with Ly6g) and macrophages (marked with CD68) in the spleen. Interestingly, IL-17A partly mediated fluoride-induced riboflavin metabolism disorder and immunotoxicity in the spleen. This work not only reveals a novel toxic mechanism for fluoride but also provides new clues for exploring the physiological function of riboflavin and for diagnosing and treating the toxic effects of fluoride in the environment.