Senora Acero features Pepito, a prominent gay character whose identity and romantic relationship are explored with dignity and complexity. The show depicts the challenges of homophobia as external forces, maintaining an empathetic and affirming stance towards his queer identity and love.

Senora Acero features Rosario, a transsexual character whose journey includes navigating societal prejudice and gaining acceptance from her mother, La Tuti. The show portrays Rosario with dignity and agency, framing obstacles as external. La Tuti's arc from initial struggle to fierce support for her daughter contributes to a net positive and affirming portrayal of transsexual identity and the importance of familial acceptance.

The series frequently depicts characters, including those involved in criminal activities, engaging with Christian (specifically Catholic) practices like prayer, confession, and invoking divine intervention. While it exposes hypocrisy among some adherents, the narrative often frames genuine faith as a source of hope, moral guidance, and a path to redemption for others, positioning the audience to distinguish between individual failings and the inherent virtues of the faith.