Ticks, as obligate blood-sucking ectoparasitic arthropods, are intricately linked to ecosystem stability. Global warming significantly impacts tick survival and reproduction, affecting individual behavior, population physiology, adaptive evolution, and altering the likelihood of tick-borne pathogen transmission, thereby posing a threat to public health. To explore the impact of elevated temperatures on ticks and the associated protein regulatory mechanisms, a quantitative proteomic analysis using the DIA technique was conducted on the ovary, salivary gland, and midgut of H. longicornis exposed to high temperatures. The findings revealed substantial changes in protein expression under high temperature stress, impacting various crucial biological processes. Specifically, Ubiquitin carboxyl-terminal hydrolase (UCH) exhibited upregulation across all tested high temperatures. Knocking down UCH through RNA interference resulted in decreased heat tolerance in H. longicornis, underscoring the essential role of UCH in maintaining normal physiological functions of ticks in high-temperature environments. This study sheds light on the physiological mechanisms employed by ticks to adapt to climate change and aids in the prevention and management of tick-borne diseases.