In the lining of water conveyance tunnels, the expansion joint is susceptible to leakage issues, significantly impacting the long-term safety of tunnel operations. Polyurea is a type of protective coating commonly used on concrete surfaces, offering multiple advantages such as resistance to seepage, erosion, and wear. Polyurea coatings are applied by spraying them onto the surfaces of concrete linings in water conveyance tunnels to seal the expansion joint. These coatings endure prolonged exposure to environmental elements such as water flow erosion, internal and external water pressure, and temperature variations. However, the mechanism of polyurea coating’s long-term leakage prevention failure in tunnel operations remains unclear. This study is a field investigation to assess the anti-seepage performance of polyurea coating in a water conveyance tunnel project located in Henan Province, China. The testing apparatus can replicate the anti-seepage conditions experienced in water conveyance tunnels. An indoor accelerated aging test plan was formulated to investigate the degradation regular pattern of the cohesive strength between polyurea coating and concrete substrates. This study specifically examines the combined impacts of temperature, water flow, and water pressure on the performance of cohesive strength. The cohesive strength serves as the metric for predicting the service lifetime based on laboratory aging test data. This analysis aims to evaluate the polyurea coating’s cohesive strength on the tunnel lining surface after five years of operation.