A member of the Ferrara branch of the influential Bentivoglio family of Bologna, he was the younger son of marchese Cornelio Bentivoglio and Isabella Bendidio. After studying at the universities of Ferrara and Padua, where in 1598 he received a doctorate utrique jure— in both civil and canon law— he returned to Ferrara, to the humanistic studies that honed his elegant writing style. There Pope Clement VIII, on a visit to the city that had recently fallen under direct papal control at long last, made him his private chamberlain, and he returned with Clement to Rome.
Under Clement's successor, Pope Paul V, he was elected titular archbishop of Rhodes, 14 May 1607, with a dispensation for being three months shy of the canonical age and not having yet received the sacred orders, in order to give him appropriate credentials as nuncio in Flanders, (1 June 1607-24 October 1615). Three topics occupied almost all his time: the struggle over the Jülich-Cleves inheritance, which was set to ignite the Thirty Years War, the flight of the prince de Condé from France in objection to Henri IV's divorce and remarriage, and the degree of toleration for Catholics in England and Ireland under James I. His correspondence reveals Bentivoglio as "the skilled diplomatist, polished by constant intercourse with the most refined society, as well as the mature observer," according to Ludwig Pastor.
Afterwards he was nuncio in France (9 July 1616). On his return to Rome in 1621 he bought Scipio Borghese's new palazzo on the Quirinale and was created cardinal in June of that year and entrusted by Cardinal Richelieu and Louis XIII with the management of French affairs at the papal court, a position he retained until 1641, when Pope Urban VIII, his intimate friend, appointed him to the suburbican see of Palestrina. Filippo Baldinucci, the biographer of Claude Lorrain, asserts that Cardinal Bentivoglio launched the artist's career by purchasing two landscapes by him, which brought the artist to the attention of Urban VIII. On 22 June 1633, Cardinal Bentivoglio was one of the signers of the papal condemnation of Galileo. An able writer and skilful diplomat, Bentivoglio was marked out as Urban's successor, but he died suddenly after the opening of the 1644 conclave. He is buried in the church of San Silvestro nel Quirinale, Rome.
Bentivoglio took Girolamo Frescobaldi with him to Brussels as his household composer. He commissioned his portrait from Anthony van Dyck (illustration); his portrait bust was sculpted by François Duquesnoy "Il Fiammingo", a Franco-Flemish sculptor working in Rome
The complete edition of his works was published at Venice in 1668.