Yang Bo, Wang Feiqing, Yang Xu, Yuan Xiaoshuang, Yang Yuting, Chen Xiaoxu, Tian Tingting, Chen Fa, Tang Dongxin, He Zhixu, Liu Yang, Li Yanju
Clinical Medical Research Center, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, No. 71 Bao Shan North Road, Yunyan District, Guiyang, 550001, Guizhou Province, China.
Academy of Medical Engineering and Translational Medicine, Tianjin University, Tianjin City, China.
Biol Trace Elem Res. 2025 Jan 17. doi: 10.1007/s12011-024-04503-y.
Chronic fluorosis is often accompanied by neurological symptoms, leading to attention, memory and learning ability decline and causing tension, anxiety, depression, and other mental symptoms. In the present study, we analyzed the molecular mechanisms of SIRT1-BDNF regulation of PI3K-AKT, MAPK, and FOXO1A in F-treated BV2 cells. The cytotoxic effect of sodium fluoride (NaF) on BV2 cells was assessed using Cell Counting Kit-8 (CCK-8), crystal violet, and 5-ethynyl-2'-deoxyuridine (EdU) staining. Cell cycle progression and apoptosis were evaluated through flow cytometry and western blotting. Reactive oxygen species (ROS) levels, oxidative stress, and inflammatory markers were measured by ROS staining, microplate reader assays, and western blotting. The role of SIRT1 in fluoride-induced toxicity for glial cells was determined using the SIRT1 activator SRT1720. The experiments demonstrated that NaF was toxic to BV2 cells, inhibited their proliferative ability, halted their cell cycle progression, triggered cellular apoptosis, promoted cellular oxidative stress (detected by ROS, SOD, MDA, GSH-Px, T-AOC) and associated protein NQO-1 and HO-1, and elevated inflammatory mediator associated protein IL-1and IL-6 expression). The fluoride-exposed groups had reduced SIRT1, BDNF, TrkB, PI3K, AKT, and MAPK protein expression levels, and increased FOXO1A protein expression. SRT1720 mitigated the harmful effects of NaF, stimulated cell proliferation and cell cycle progression, decreased apoptosis, reduced oxidative stress and inflammatory factors, elevated SIRT1, BDNF, TrkB, PI3K, AKT, and MAPK protein levels, and suppressed FOXO1A protein expression. The results indicate that NaF potentially harms glial cells by suppressing SIRT1 activation, and SIRT1 significantly mitigated the damage. Furthermore, the SIRT1 signaling pathway might regulate the nerve damage caused by fluoride poisoning and may be a protective factor in treating fluoride-induced brain injury.
慢性氟中毒常伴有神经症状，导致注意力、记忆力和学习能力下降，并引发紧张、焦虑、抑郁等精神症状。在本研究中，我们分析了SIRT1-BDNF对氟处理的BV2细胞中PI3K-AKT、MAPK和FOXO1A的调控分子机制。使用细胞计数试剂盒-8（CCK-8）、结晶紫和5-乙炔基-2'-脱氧尿苷（EdU）染色评估氟化钠（NaF）对BV2细胞的细胞毒性作用。通过流式细胞术和蛋白质印迹法评估细胞周期进程和细胞凋亡。通过ROS染色、酶标仪检测和蛋白质印迹法测量活性氧（ROS）水平、氧化应激和炎症标志物。使用SIRT1激活剂SRT1720确定SIRT1在氟诱导的神经胶质细胞毒性中的作用。实验表明，NaF对BV2细胞有毒性，抑制其增殖能力，阻止其细胞周期进程，引发细胞凋亡，促进细胞氧化应激（通过ROS、SOD、MDA、GSH-Px、T-AOC检测）及相关蛋白NQO-1和HO-1，并升高炎症介质相关蛋白IL-1和IL-6的表达。氟暴露组的SIRT1、BDNF、TrkB、PI3K、AKT和MAPK蛋白表达水平降低，而FOXO1A蛋白表达增加。SRT1720减轻了NaF的有害影响，刺激细胞增殖和细胞周期进程，减少细胞凋亡，降低氧化应激和炎症因子，提高SIRT1、BDNF、TrkB、PI3K、AKT和MAPK蛋白水平，并抑制FOXO1A蛋白表达。结果表明，NaF可能通过抑制SIRT1激活对神经胶质细胞造成损害，而SIRT1可显著减轻这种损害。此外，SIRT1信号通路可能调节氟中毒引起的神经损伤，可能是治疗氟诱导脑损伤的保护因子。
