Aircraft noise pollution is getting increasingly severe due to striding development of worldwide air transport. In high-density cities, a large number of residential areas are exposed to airport routes where flight-over noise poses a serious threat to public health. However, most existing research is limited to the thresholds for annoyance lacking in-depth exploration into the psychophysiological mechanisms that induce health responses. This study aims to examine the health effects of aircraft noise on both subjective evaluation and objective physiological indicators in a laboratory setting. Aircraft noise samples were collected in the under-route open spaces and their effects on annoyance and physiological feedback, including Skin Conductance Response (EDA) and Heart Rate Variability (HRV), were examined using listening experiment and ErgoLAB electrophysiological response monitoring. The study analysed the relationship between psychophysiological responses and subjective annoyance. The psychoacoustic parameters that induced emotion-related feedback were also identified. The results are expected to provide indicators to assess and improve the health of under-route communities.