Background: For mild acne vulgaris that does not respond to conventional therapy modalities and severe nodulocystic acne vulgaris, oral isotretinoin is known to be the most effective treatment. It is the only form of therapy that affects the full etiology of acne. Objective: This study aimed at investigating how isotretinoin affected the levels of prolactin and androgens in the blood of females with acne vulgaris. Patients and methods: Fifty adult females with acne vulgaris, between puberty and menopause, who were candidates for isotretinoin treatment, were included in this study. Patients were chosen from the outpatient dermatology clinics. Results: After receiving isotretinoin treatment, there was a statistically significant drop in prolactin levels; the mean difference was 5.33 and the percentage change was 25.4 percent. Following treatment, there was a statistically significant rise in both DHEAS and free testosterone levels in the cases, with mean differences of 0.58 and -0.22 and percentage changes of 21.5 percent and 27.2 percent respectively. Conclusion: According to this study, isotretinoin has been successfully used to treat cases of acne vulgaris. It can lower prolactin levels while increasing DHEAS and free testosterone; two key factors that contribute to acne etiology.