Investigating variations of rock fracture toughness after high-temperature thermal fatigue is greatly significant for developing deep geothermal resources. This study investigated the mode I fracture toughness of gabbro after high-temperature fatigue treatment by heating a cracked straight-through Brazilian disc (CSTBD) gabbro specimen to 200, 400, 500, and 600 °C, followed by 100 thermal cycles. Additionally, acoustic emission (AE) and digital image correlation (DIC) were used to monitor the failure process of gabbro. The results showed that the peak load, fracture toughness, and fracture energy of gabbro decreased with the increased temperature and number of cycles, which was most significant at 500 and 600 °C. Under mode I loading, cracks developed along the loading direction and connected with the prefabricated crack. Additionally, the regions with higher von Mises equivalent strain (Evm) values were concentrated at the loading points and prefabricated crack of the rock sample. The degree of bending of the primary cracks increased, the number of secondary cracks increased, and the Evm values changed significantly as the temperature increased. The phase transformation of plagioclase feldspar at 450 °C and the oxidation of iron- and magnesium-containing pyroxene at 600 °C changed the internal structure of the rock. Cyclic treatment further exacerbated the thermal damage, developing microcracks within the rock and reducing fracture toughness significantly.