Adolescents, as a large group in the world, face many physical changes and psychological evolutions in their puberty period. If enough attention is not paid to such changes, negative effects on their health and knowledge may be induced. Thus, it is very important to hold health education approperiate for their needs using new educational methods and confident sources. The main goal of this study is to explore the impact of puberty health education on the anxiety of girls. Itis a quasi-experimental study using clustered sampling which was done on 159 girls from two high schools in Tehran divided into two experimental (N=86) and control (N=73) groups. Then,using a systematic educational plan revised by the researcher and expert panel from Department of Midwifery, all the students and their parents in the experimental group were instructed. Data were gathered by demographic questionnaire and Spielberger Scale. Questionnaires were completed by students in three phases including before, after, and three months after the end of the educational program. Data analysis was performed by paired t-test, independent t-test, Chi square, and multivariate tests. Mean anxiety scores in the experimental and control groups were 90.45 and 85.36 before the education, 78.79 and 85.49 at the end of the education, and 78.46 as well as 87.33 3 months later, respectively.Anxiety scores were statistically different post-intervention (p<0.001) and three months later(p<0.001). Puberty health education programs could reduce anxiety in female adolescents.