Xin Jinge, Chen Yu, Huang Yongmei, Sun Ning, Peng Weiqi, Lai Chunxiao, Yang Ruanruan, Chen Wenqing, Duan Lixiao, Wang Dandan, He Yuhao, Bai Yang, Ni Xueqin, Wang Hesong
Department of Gastroenterology, Baiyun District People's Hospital of Guangzhou, Guangzhou, Guangdong, China.
Animal Microecology Institute, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, China.
Theranostics. 2025 Jun 18;15(15):7242-7269. doi: 10.7150/thno.116567. eCollection 2025.
Chronic exposure to high-fluoride drinking water impairs intestinal structure and function, potentially damaging extraluminal tissues via the gut-organ axis. However, how lifelong exposure to naturally occurring moderate-to-high-fluoride water affects intestinal cells and their underlying mechanisms remain unclear. Single-cell RNA sequencing identified cellular heterogeneity and candidate risk genes in the mouse ileum after 56 weeks of 50-ppm fluoride exposure. Cellchart was employed to analyze fluoride-altered cell communication patterns, and gut bacterial richness was ablated using broad-spectrum antibiotics to validate high fluoride-disrupted intercellular signaling pathways. Fluoride exposure disrupted enterocyte trans-differentiation, affected metabolic health by restricting nutrient absorption, and activated antibacterial activity in enterocytes at the villus base. Downregulation of genes associated with rapid goblet-cell emptying and transmembrane mucin 3 in goblet cell impairs mucus and glycocalyx formation. Antimicrobial peptides, lectins, and lysozymes were reduced in fluoride-exposed Paneth and goblet cells. Fluorescence hybridization demonstrated bacterial invasion of the epithelium following mucus barrier damage. Immunologically, fluoride-exposed T cells exhibit high scores for apoptosis, cell cycle suppression, inflammation, and high gut-homing gene expression. Fluoride exposure promoted somatic hypermutation and affinity selection in B-lineage cells while expanding plasma cells with high developmental potential. Ligand-receptor analysis revealed that activated enterocytes presented antigens to T cells via the MHC-II L-R signaling pathway, triggering downstream responses such as upregulating proinflammatory factors and cytotoxic molecules, and remodeling B-lineage cells. Broad-spectrum antibiotics depleted gut microbiota, reducing fluoride-induced gut microbial overgrowth and suppressing MHCII signaling in enterocytes and T/B cell activation-thereby decreasing proinflammatory cytokines and immunoglobulins. High-fluoride exposure disrupts the intestinal mucus barrier and gut microbiota homeostasis, leading to bacterial invasion of the epithelium that activates MHC-II signaling in absorptive enterocytes. Upregulated MHC-II signaling triggers intestinal immune cell activation and inflammation. These results reveal new intercellular interactions and communication hubs in intestinal cells under fluoride exposure.
长期暴露于高氟饮用水会损害肠道结构和功能，可能通过肠-器官轴损害肠外组织。然而，终生暴露于天然存在的中高氟水中如何影响肠道细胞及其潜在机制仍不清楚。单细胞RNA测序确定了在50 ppm氟暴露56周后小鼠回肠中的细胞异质性和候选风险基因。使用Cellchart分析氟改变的细胞通讯模式，并使用广谱抗生素消除肠道细菌丰富度，以验证高氟破坏的细胞间信号通路。氟暴露破坏肠上皮细胞转分化，通过限制营养吸收影响代谢健康，并激活绒毛基部肠上皮细胞的抗菌活性。杯状细胞中与快速杯状细胞排空和跨膜粘蛋白3相关的基因下调会损害黏液和糖萼的形成。在暴露于氟的潘氏细胞和杯状细胞中，抗菌肽、凝集素和溶菌酶减少。荧光杂交显示黏液屏障受损后细菌侵入上皮。在免疫学上，暴露于氟的T细胞在细胞凋亡、细胞周期抑制、炎症和高肠道归巢基因表达方面得分较高。氟暴露促进B系细胞中的体细胞超突变和亲和力选择，同时扩增具有高发育潜力的浆细胞。配体-受体分析表明，活化的肠上皮细胞通过MHC-II L-R信号通路将抗原呈递给T细胞，触发下游反应，如上调促炎因子和细胞毒性分子，以及重塑B系细胞。广谱抗生素消耗肠道微生物群，减少氟诱导的肠道微生物过度生长，并抑制肠上皮细胞中的MHCII信号和T/B细胞活化，从而减少促炎细胞因子和免疫球蛋白。高氟暴露破坏肠道黏液屏障和肠道微生物群稳态，导致细菌侵入上皮，激活吸收性肠上皮细胞中的MHC-II信号。上调的MHC-II信号触发肠道免疫细胞活化和炎症。这些结果揭示了氟暴露下肠道细胞中新的细胞间相互作用和通讯枢纽。
