Safflower is known for its effectiveness in preventing arteriosclerosis and is rich in linoleic acid. It contains various compounds including flavonoids, alkaloids, and phenolic acids, which contribute to its efficacy in cardiovascular metabolism. Hydroxysafflor yellow A (HSYA) found in safflower shows potential in treating cerebrovascular diseases by reducing neurological deficits in a cerebral infarction model. In modern society, many individuals experience stress, and humanities therapy is considered positive for relieving stress induced by emotional labor. The autonomic nervous system (ANS) regulates internal organs, and stressful situations can disrupt this balance. We investigated the effects of inhaling safflower extract and attending humanities lectures on autonomic nervous system (ANS) activity using transcranial Doppler ultrasonography. The study results showed an increase in the activity of the sympathetic and parasympathetic nervous systems after inhaling safflower extract and attending humanities lectures. Brain activity increased, indicating improved concentration and immersion, while stress on the brain remained relatively low. This study provides a foundation for understanding the influence of safflower extract on cognitive function and the autonomic nervous system, suggesting the need for further research on safflower extract and the autonomic nervous system.