This study investigated the potential mechanism of action of tea polyphenols (TPs), one of the major active ingredients in tea, to enhance heat resistance in Drosophila and the attenuating effect of heat treatment of TPs on their efficacy. The results showed that TPs were able to prolong the average survival time of Drosophila under high-temperature stress (p < 0.05), but the effect of TPs in prolonging the survival time of Drosophila melanogaster was significantly reduced (p < 0.05) with increasing TP heat-treatment time until it disappeared. The composition of TPs changed after heat treatment. It was also shown that the weakening of the effect of TPs in improving the heat tolerance of Drosophila was related to the decrease in the content of catechins and phenolic acids in their fractions as well as with the increase in the content of laccase. Transcriptomic analysis showed that the effect of TPs on heat tolerance in Drosophila melanogaster was closely related to the longevity regulation pathway, the neuroactive ligand-receptor interaction signaling pathway, and the drug metabolism-cytochrome P450 pathway. Metabolomics analysis showed that the effect of TP intervention in improving the body's heat tolerance was mainly related to amino acid metabolism and energy metabolism. However, thermal processing weakened the relevance of these transcriptomes and metabolomes. The present study reveals the mechanism of action by which heat-treated TPs affect the body's heat tolerance, which is important for the development and utilization of the heat-protection function of tea.