In the 10-minute video, reviewed by the counter-terrorism company IntelCenter and posted Friday, Zawahiri tells Pakistanis to "rise up in the face of this bribe-taking government … these treacherous generals," Voice of America reported.
Zawahiri urged Pakistanis to follow the example of Muslims in Egypt and Tunisia by revolting against their government.
He said Pakistan's government has "spilled the blood of Muslims in Afghanistan and Pakistan to fill the pockets of its leadership" and "will not fight in your defense."
Zawahiri also said in the video al-Qaida will not release Warren Weinstein, an American development expert who had been living in Pakistan and was kidnapped, until the terrorist group's demands are met.
IntelCenter said the video message appears to have been produced in February.
VOA said al-Qaida released a video last month expressing support for the uprising against Syrian President Bashar Assad and urged Muslims in neighboring countries to aid anti-government rebels in Syria.