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Homophobic bullying is a major social issue, especially in the school settings. This may be particularly common in many Western countries but it appears globally. Bullying causes both short and long-term problems hence its prevention must be seen as a relevant and urgent step in educational settings. Psycho-educational programs should be promoted in order to help eliminate school-children's prejudices and bias about gay and lesbian peers, homophobic bullying. 191 school-children ( n  = 101 females, n  = 90 males), aged 12-14 years old, attending a secondary school in Foggia (Italy) were recruited and assessed at baseline (T0) and 2 months (T1) after receiving a systematized, repeated psycho-education aimed to promote their awareness on sexual variations, and reduce homophobic prejudices and stereotypes. They were assessed for their empathy quotients, their knowledge of gender- related stereotypes, homophobic attitudes, anger, emotional regulation, before (T0) and after the intervention (T1) in a standardized manner. The psychoeducational program significantly reduced homophobia levels towards gays (-9.38%) and lesbians (-5.42%,) as well as improved emotional adjustment (+25.9%) and pro-sociality (+3.85%) among school-children (0.0486≤ all p  < 0.0001). Also, a statistically significant improvement of empathy (+11.3%) and levels of state anger (+0.35%) has been significantly reported. This study reports on the benefits of a repeated, systematic, prospective psycho-educational intervention conducted in the school-setting leading to an improvement of empathy, pro-sociality, emotional regulation and a reduction of homophobic attitudes and anger among students.