"God Control" features a graphic nightclub shooting, calling people to "wake up" to the reality of arms ownership and use.
Madonna stepped into the gun violence debate on Wednesday with a music video containing graphic dramatisations of a mass shooting.
Called "God Control," the disco beat song and video depicts a gunman opening fire in a crowded nightclub or bar and images of actors playing dancers who fall to the ground, bleeding, or try to flee.
A message before the video reads, "The story you're about to see is very disturbing. It shows graphic scenes of gun violence. But it's happening every day. And it has to stop."
"We need to wake up," Madonna sings.
The video also uses TV footage of demonstrators calling for stricter gun control, including the 2018 "March For Our Lives" in Washington and other US cities.
Florida nightclub shooting
While the music video does not refer to any one incident, it recalls the June 2016 shooting inside Pulse the gay nightclub in Florida, in which 49 people were killed and 53 injured.
Madonna, 60, said the video was designed to shock.
"Seeing the reality, and the brutality of things makes you wake up," the singer told People magazine on Wednesday.
"This is really happening. This is what it looks like. Does it make you feel bad? Good, 'cause then maybe you will do something about it," she said.
The United States has seen hundreds of mass shootings in schools, workplaces, places of worship and leisure in recent years but stricter gun control legislation has proven difficult to pass in the US Congress.
In a message on her social media accounts, Madonna urged people to "insist on common-sense gun safety legislation" and directed fans to gun control organisations they could support.
The "God Control" video had been viewed over one million times on YouTube since its release on Wednesday.