The ecological environment of the Hexi Corridor in China is fragile owing to its unique geographical conditions. Since the 21st century, the construction of highway projects in the Hexi Corridor has increased with the implementation of China’s western development policy. The impact of highway construction on vegetation along roads is uncertain and requires attention. In this study, the normalized difference vegetation index (NDVI) was obtained based on remote sensing data, and the correlations between the vegetation index, climate changes, and highway construction from 2000 to 2019 were analyzed. The results showed that the NDVI of the Hexi Corridor showed a significant increasing trend, and the spatial difference was obvious, which was largely controlled by climatic conditions. Generally, the vegetation index was negatively correlated with temperature, but positively correlated with precipitation, and the correlation between the NDVI and precipitation was greater than that of temperature. The impact of highways on vegetation is a long-term process. In the eastern region of the Hexi Corridor, where the ecological environment is better, highway construction promoted vegetation development. However, in the western region with high desertification, the damage caused by highway construction to vegetation was serious, and the recovery rate of degraded vegetation was slow after the completion of highway construction. Although the vegetation development in the Hexi Corridor was mainly affected by precipitation, in the western region, highway construction was the main factor controlling vegetation disturbance within a certain range on both sides, and its contribution to vegetation disturbance reached 60%. The research results can improve understanding of the disturbance effect of highway construction on vegetation in arid areas.