Abstract Introduction Self-medication is a common practice these days and is influenced by many factors, such as education, family, society, availability of medications and exposure to advertisements. This practice should be done responsibly for relief and treatment of brief and non-serious health conditions. Its irresponsible use can create risks to the individual’s health. Objectives This work aims to evaluate the knowledge about self-medication on higher education students, and on a village population located at the interior of the Portugal central region, in order to verify if health education and school level influence the behaviours how self-medication is done. Methodology An observational study was carried out at the Coimbra Health School (ESTeSC), and at a Pharmacy in a rural village, with a questionnaire done between March and May 2019. In this study 430 individuals contributed, of which 223 were enrolled in ESTeSC and 207 were Pharmacy’s customers. Results At the sample, female students (88.8%) predominated, with an average age of 20 years old. From the results collected at the Pharmacy’s customers, female individuals predominated (65.7%). From the overall, 75.8% indicated that they self-medicated in the last 6 months (170 individuals from ESTeSC and 156 from pharmacy) mainly for Flu/Cold symptoms (38.8%). Analgesics and antipyretics predominates self-medication with 36.3% and 41.06% (ESTeSC and Pharmacy, respectively). They further stated that self-medication is acceptable in some circumstances. Conclusion We can conclude that regarding to self-medication practice, individuals increasingly look out to treat their symptoms without seeking medical advice, as it is faster and cheaper. Therefore, pharmacy professionals must educate population about benefits and risks of self-medication. These professionals have an important role at the self-medication risks prevention, sharing information, therapeutic guidance and population education.