Laser damage resistance of films is the key to the performance and durability of high-power laser systems. High temperature, however, often leads to a certain degree of decline in film properties. Here, aiming to explore the mechanism of laser damage under high temperatures, Nb2O5 films were prepared using sol–gel method with NbCl5 as precursor and citric acid as additive. The effect of annealing temperatures in optical performance, microstructure, surface morphologies, and chemical composition of films were studied. Further investigation was carried out of the laser-induced damage threshold (LIDT) under different in situ high temperatures of the as-deposited films. The results showed that the films had prominent optical transmittance and high LIDT. Under 293 K, the LIDT of the film was the highest of 24.8 J/cm2. The increase of temperature brought down the LIDT of the films. It was attributed to the occurrence of oxygen vacancies, the sharp increase of temperature, and rise of defects and destruction of network structure. In this study, even though the LIDT of the film decreased a lot at 523 K, it was still high up to 14.1 J/cm2, representing the great potential for applications in authentic high-temperature environments.