Aim: To explore the relation between male testosterone (T) and androgen binding protein (ABP) with fluoride exposure in drinking water. Method: Cross-sectional study was conducted in 7 villages of a county in Henan Province as investigation points including 2 high-fluoride villages, 2 villages with water improvements and 3 control villages for collecting drinking water in all investigation points. Males who were 18 to 50 years old and born in the [abovementioned] villages were selected from investigation points as investigation subjects through group sampling. Morning urine and fasting venous blood were collected respectively. Fluoride content in drinking water and urine was determined by fluoride ion selective electrode method; serum ABP level was determined by ELISA method and serum T was determined by chemiluminescence immunoassay. Result: Fluoride concentration of drinking water for high-fluoride group is (2.44 ± 1.88) mg/L which is higher than (0.37 ± 0.15) mg/L of control group and higher than (0.36 ± 0.30) mg/L of improved-water group (F = 12.289, P < 0.001). Urinary fluoride concentration of high-fluoride group is (2.49 ± 1.40) mg/L which is higher than (1.04 ± 0.49) mg/L of control group and higher than (1.38 ± 0.67) mg/L of improved-water group (F = 71.563, P < 0.001); [the urinary fluoride concentration] of improved-water group is also higher than that of control group (P < 0.05). Serum ABP content of high-fluoride group is (16.01 ± 10.83) nmol/L which is lower than (27.94 ± 31.90) nmol/L of control group and lower than (22.42 ± 28.12) nmol/L of improved-water group (F = 28.807, P < 0.001). Serum T content of control group, improved-water group and high-fluoride group is (4.31 ± 1.30), (4.42 ± 1.37) and (4.74 ± 2.17) nmol/L [respectively]. The difference was of no statistical significance (F = 0.268, P = 0.765). In control group and improved-water group, negative correlation exists between serum T content and age (? = -0.238, -0.262, P < 0.05 for both groups). Conclusion: Environmental fluoride exposure may influence serum ABP level in males.