Endemic fluorosis is a serious problem in public health. Previous studies have indicated that patients with thyroid goiters usually live in fluoride-affected areas. However, the mechanism of goitrogenesis caused independently by fluoride is still unclear. The principle objective of this study was to investigate the possible roles of nitric oxide (NO) and vascular endothelial growth factor (VEGF) in the genesis of fluoride-induced nodular goiters. Eighty SD rats (40 males and 40 females) at the age of 4 weeks were used to establish animal models via fluoride-supplemented drinking water. These rats were randomly divided into four groups of 20. Group 1 was used as the control and were given deionized water. Group 2 (LF), group 3 (MF), and group 4 (HF) were given deionized water containing 50mg/L, 100mg/L, and 200mg/L of sodium fluoride (NaF), respectively. Thyroid samples were collected on day 150. Pathological observation was performed to evaluate structural changes in the thyroid gland. The expression of VEGF mRNA in the thyroid glands was assessed by reverse transcriptional PCR. The serum NO level was analyzed by spectrometric methods. In addition, immunohistochemistry was conducted to evaluate expression and deposition of VEGF in the thyroid gland. The results showed that the average relative weight of the thyroid glands of rats in the fluoride-treated groups was significantly higher than that in control rats (p<0.05). The proliferation and dilatation of capillary blood vessels, enlarged follicles with excessive colloid, and obvious nodules were found in the thyroid glands of fluoride-treated rats. Compared to the control group, the expression of VEGF mRNA in the thyroid gland and the serum NO levels in the fluoride-treated groups were significantly increased (p<0.05). Furthermore, the deposition of VEGF in epithelial and follicular cells of the thyroid gland was significantly higher in fluoride-treated groups than in the control group. These results suggested that abnormal expression of VEGF induced by fluoride can lead to the proliferation of vascular endothelial cells in the thyroid gland. Accordingly, VEGF oversecreted locally by vascular endothelial cells might contribute to the proliferation of epithelial and follicular cells, resulting in the formation of hyperplastic nodules and enlargement of the thyroid gland. Furthermore, we proposed that there might be a positive feedback mechanism between NO and VEGF expression in fluoride-induced goiter formation. It was concluded that angiogenic and vasodilative factors such as VEGF and NO must be involved in fluoride-induced thyroid goitrogenesis.