The California lawmaker, who played herself, blasts Alec Baldwin's TV character, Jack Donaghy, as an "economic war criminal" in a mock MSNBC interview as Donaghy watches from his Manhattan office.
Pelosi also pokes fun at Congress during the 10-second cameo.
"Jack Donaghy is an economic war criminal. If the Democratic Party controlled Congress, I would see that he was punished in the worst way possible, by having to come down here and listen to us," Pelosi said.
Last night's episode also featured guest stars Julianne Moore, Salma Hayek and Conan O'Brien, as well as Ice-T and Richard Belzer from "Law & Order: Special Victims Unit," but Pelosi was the only elected official in sight.
The Emmy-award winning "30 Rock," which went off the air after seven seasons, has had its share of high-profile political guests, including former Vice President Al Gore and former Secretary of State Condoleezza Rice.
The show's star, Tina Fey, has tried her hand at faux politics, famously portraying former vice presidential candidate and former Alaska Gov. Sarah Palin on "Saturday Night Live."
Asked Monday if Fey would ever consider a career in politics now that her series was ending, she quickly answered, "nooooo."