To reveal the influencing factors and changing rules for the hydrothermal interaction process of highway subgrade, the field measurements of Shiwei-Labudalin Highway in Inner Mongolia, China was conducted for 3 years, based on which the freezing-thawing rules and water content changing characteristics were analyzed. The main results show the subgrade presents a frequent freezing-thawing alternation, and the water content of subgrade exhibits an obvious seasonal alternation. The subbase has the maximum water content, while the base has the minimum water content. The change of water flux is concentrated in the thawing period and consistent with the change of temperature gradient. The subbase layer has the most active water flux due to the heat absorption and impermeability of pavement that easily causes the water accumulation in this layer. Therefore, the prevention and treatment for the freezing-thawing disease should be started from heat insulation and water resistance.