Wang Feiqing, Chen Fa, Song Wen, Li Yanju, Wu Haiyan, Tian Tingting, Tian Mengxian, Tang Dongxin, Liu Yang
Clinical 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, 300072, China.
Biol Trace Elem Res. 2024 Sep 17. doi: 10.1007/s12011-024-04381-4.
Fluorosis is a worldwide public health problem, in which the heart is an important target organ. However, studies on its toxicological mechanism in embryonic development are limited. This study assessed the toxicity of sodium fluoride (NaF) toward zebrafish embryos. We determined the mortality, hatching rate, phenotypic malformation, heart function, and morphology of zebrafish embryos after exposure to NaF. Subsequently, the molecular mechanism was revealed using high-throughput RNA sequencing analysis. The expression levels of key genes for heart development were detected using quantitative real-time reverse transcription PCR. The 50% lethal concentration (LC50) value of NaF toward zebrafish embryos at 96 h post-fertilization was 335.75 mg/L. When the concentration of NaF was higher than 200 mg/L, severe deformities, such as pericardial edema, yolk sac edema, spine curvature, shortened body length, reduced head area, and eye area, were observed. The heart rate of the embryos exposed to NaF decreased in a dose-dependent fashion. The distance between the sinus venosus and bulbus arteriosus was significantly increased in the NaF-exposed group compared with that in the control group. The stroke volume and cardiac output decreased significantly in the NaF groups. Compared with the control group, the expression levels of Gata4, Tbx5a, Hand2, Tnnt2c, Nppa, and Myh6 were significantly increased in the NaF-treated group. Through transcriptome sequencing, 1354 differentially expressed genes (DEGs) were detected in the NaF (200 mg/L) treated groups, including 1253 upregulated genes and 101 downregulated genes. Gene ontology functional analysis and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the DEGs showed that cardiac-related pathways, such as actin cytoskeleton regulation, Jak-Stat, PI3k-Akt, and Ras, were activated in the NaF-exposed group. This study revealed the underlying mechanism of fluoride-induced cardiac morphological and functional abnormalities and provides clues for the clinical prevention and treatment of fluorosis.
氟中毒是一个全球性的公共卫生问题，其中心脏是一个重要的靶器官。然而，关于其在胚胎发育中的毒理学机制的研究有限。本研究评估了氟化钠（NaF）对斑马鱼胚胎的毒性。我们测定了暴露于NaF后斑马鱼胚胎的死亡率、孵化率、表型畸形、心脏功能和形态。随后，使用高通量RNA测序分析揭示分子机制。使用定量实时逆转录PCR检测心脏发育关键基因的表达水平。受精后96小时，NaF对斑马鱼胚胎的50%致死浓度（LC50）值为335.75mg/L。当NaF浓度高于200mg/L时，观察到严重畸形，如心包水肿、卵黄囊水肿、脊柱弯曲、体长缩短、头部面积减小和眼部面积减小。暴露于NaF的胚胎的心率呈剂量依赖性下降。与对照组相比，NaF暴露组的静脉窦和动脉球之间的距离显著增加。NaF组的每搏输出量和心输出量显著下降。与对照组相比，NaF处理组中Gata4、Tbx5a、Hand2、Tnnt2c、Nppa和Myh6的表达水平显著升高。通过转录组测序，在NaF（200mg/L）处理组中检测到1354个差异表达基因（DEG），包括1253个上调基因和101个下调基因。对DEG的基因本体功能分析和京都基因与基因组百科全书通路分析表明，在NaF暴露组中，肌动蛋白细胞骨架调节、Jak-Stat、PI3k-Akt和Ras等心脏相关通路被激活。本研究揭示了氟诱导心脏形态和功能异常的潜在机制，并为氟中毒的临床防治提供了线索。
