The video, which appeared on social media on Friday, tells voters that if Putin's rivals win they will take Russia into a "nightmare" future in which they are forced to co-habit with gay men.
The video stars well-known screen actors but it was unclear who published it.
It has had more than three million views on the Facebook page of pro-Kremlin political analyst Alexander Kazakov.
In it, a middle-aged man dreams of a future without Putin, who is expected to win a fourth term in the March 18 vote.
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One of Putin's election rivals, liberal candidate Ksenia Sobchak, has called for Russia to allow gay marriage -- an unlikely prospect in a socially conservative country.
Some observers have suggested that Sobchak's candidacy was part of a strategy to create a semblance of diversity in an election that Putin is all but certain to win.
She has denied being a "Kremlin project."
Sobchak spoke out against the video on Monday, condemning it as "vile".