Originally from the castle of that name in the neighborhood of Bologna, the family claimed descent from Enzio, King of Sardinia, an illegitimate son of Frederick II, Holy Roman Emperor.

Amid the faction-conflicts of the commune, on 14 March 1401, Giovanni I Bentivoglio, with the help of Gian Galeazzo Visconti, declared himself signore and Gonfaloniere di Giustizia.

The Visconti however soured on Giovanni, and he was defeated and killed on 26 June 1402 at the Battle of Casalecchio and was interred in the church of San Giacomo Maggiore.

The son of Giovanni I, Anton Galeazzo (or Antongaleazzo, c. 1385–1435), was a lecturer in civil law who briefly assumed power in Bologna in 1420, but was quickly overthrown.

In 1438 Annibale I, a putative son of Anton Galeazzo (his mother, Lina Canigiani, was said to be uncertain of the boy's paternity and the matter was decided by dice)[1], led a city revolt against the Papacy.

When Annibale returned to Bologna, the powers of government were conferred to him, a sign that the city recognized the family's political importance.

Ercole Bentivoglio, the putative father of Sante I who lived in exile in Florence and Ferrara, wrote a long poem on dietetic, greatly extolling the medical properties of cheese.