Nighttime walking is becoming a popular exercise for many middle-aged and older people in Asian countries. However, the health benefits of nighttime walking in urban areas and green spaces are still unclear. This study evaluated the physiological and psychological responses of 48 middle-aged and older people who walked 1.6 km through a green space and an urban area during daytime and nighttime. The Positive and Negative Affect Schedule (PANAS), Profile of Mood States (POMS), Perceived Restorativeness Scale (PRS), and Restorative Outcome Scale (ROS) were employed to measure the psychological responses, and pulse rate and blood pressure (SBP, DBP and MAP) were measured to evaluate the physiological responses. The results showed that the daytime green walking induced psychological improvements and lowered blood pressure (p < 0.05), while the daytime urban walking resulted in slight deterioration of all the measured parameters (p > 0.05). On the other hand, the nighttime green walking induced lowered blood pressure (p < 0.05), whilst the nighttime urban walking resulted in psychological improvements and lowered blood pressure (p < 0.05), and no significant difference was found in any measured parameter between the two nighttime walking groups. In conclusion, urban areas are noisy and irritating in the daytime, and not suitable for walking, but may become pleasurable and attractive at night. The psychological benefits of green walking may decrease at night, and nighttime walking in either an urban area or a green space may achieve similar health benefits. Therefore, we recommend that urban citizens start nighttime walking in a green space or an urban area to keep fit when the air is less polluted.