Acne vulgaris (AV) is a chronic inflammatory disease of the pilosebaceous unit and commonly occurs in adolescents and young adulthood. We investigated and compared the expressions of epidermal barrier proteins of healthy, sebaceous gland-rich (SGR) skin and AV samples by immunohistochemistry (IHC). Our results showed significant changes in expressions of proteins in samples with AV compared to SGR skin samples, which have roles in corneocyte envelope formation, desmosome organization, corneocyte desquamation, tight junction formation, barrier alarmins antimicrobial proteins. We proved the damage in skin barrier functions in AV, which also highlights the importance of appliance of the barrier-improving agents simultaneously with acne therapy to improve the effectiveness of the therapy.