In the end, the award for best director went to Guillermo Del Toro for The Shape Of Water.
He beat out Martin McDonagh for Three Billboards Outside Ebbing, Missouri, Christopher Nolan for Dunkirk, Ridley Scott for All the Money in the World and Steven Spielberg for The Post.
Twitter swiftly reacted in praise of the actress for driving home a point that had been the theme of the whole evening - that women are not treated equally in Hollywood.
Most of the female celebrities who attended the event at the Beverly Hilton wore black to show support for Time's Up, an initiative that seeks to promote equality, wage parity and inclusion and also has established a legal defense fund for victims of sexual assault.
"Me and my friends just bawled out eyes out at Oprah's speech. Then Natalie Portman was like HOLD MY BEER," tweeted on female account user.
Another tweeted: "I couldn't believe someone had to immediately follow Oprah and then Natalie Portman comes through with that."
Some of Portman's fellow actresses also chimed in with Mindy Kaling tweeting: "Yas Natalie Get Em Girl."
Only one woman has ever won the Golden Globes award for film directing in the show's 75 year history. That was Barbra Streisand who won in 1984 for her film Yentl.
The musical movie also won the Golden Globe for best picture.