Susa Valley, located in the Italian Western Alps, has served as a meeting point for cultural, spiritual, and commercial exchange for a long period of history. The valley’s role as one of the main connecting routes between south and southwestern Europe resulted in its acquisition of a rich traditional ecological knowledge. However, like other Italian mountainous valleys, this valley has suffered from abandonment and depopulation in the past 50 years. Our study aims to investigate the current ethnobotanical medicinal knowledge in the valley and to compare our findings with a study conducted over 50 years ago in the same area. In 2018, we conducted 30 in-depth semi-structured interviews on medicinal plants and food-medicines used in the Susa Valley. We documented 36 species, of which 21 species were used for medical purposes and 15 species were used as food-medicine. The comparison with the previous study on medicinal herbs conducted in 1970 in the valley demonstrated a significant decrease in both the knowledge and use of medicinal plants, which could be attributed to socioeconomic, cultural, and possibly environmental changes that occurred in the past half-century. Our study highlights several promising species for future use as nutraceuticals, food, and medicinal products, such as Taraxacum officinale, Urtica dioica, and Artemisia genipi.