He warned that Iran will use its right to reciprocate the enemies' hostile moves in a way that they will regret their deeds.
US President Donald Trump has used the protests to repeatedly bash the Iranian government and warned it against any human rights abuses, building on his crusade against the 2015 nuclear deal that Iran signed with world powers under Trump's predecessor, Barack Obama.
Nine more people, including an 11-year-old boy, have been killed as anti-government protests continued across Iran on Tuesday.
Rowhani has tried to play down the unrest, which began over economic grievances in second city Mashhad last Thursday but quickly turned against the Islamic regime as a whole with chants of "Death to the dictator".
Nine people have been killed overnight amid nationwide protests and unrest in Iran.
The highest-ranking USA official's direct expression of support for rioters in recent protests in Iran and the supportive voices by a number of other governments and Western media outlets reveal that the U.S. had plans for inciting a sedition inside Iran, Brigadier General Massoud Jazayeri said on Monday.
In the latest development, state TV reported that six rioters were killed during an attack on a police station in the town of Qahdarijan when they tried to steal guns from the facility.
Iranian President Hassan Rouhani has addressed the nation for the first time since protests started across the country four days ago.
"This is nothing", Mr Rouhani said.More news: LG unveils world's first 88-inch 8K OLED display
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It was unclear if this was the same fatality as that reported by Iran's semi-official Mehr news agency on Monday. Iran has seen its largest anti-government protests since the disputed presidential election in 2009. Hundreds of demonstrators have been arrested.
"Iran is failing at every level despite the awful deal made with them by the Obama administration", Mr Trump said, referring to the nuclear pact agreed under his predecessor Barack Obama.
'We regret the loss of life that has occurred in the protests in Iran, and call on all concerned to refrain from violence and for global obligations on human rights to be observed, ' he said.
Both apps were used by activists to organise the demonstrations and share images and information.
"The government will show no tolerance for those who damage public properties, violate public order and create unrest in the society", Rouhani added.
The Iranian foreign ministry had also on Saturday strongly deplored the recent comments by Trump and State Department Spokeswoman Heather Nauert in support of the recent economic protests in Iran, describing them as "cheap, worthless and invalid".
"It's clear there's a kind of discontent that's sweeping Iran over broad issues".
In a statement on Twitter, the ministry cautioned that Iran was seeing "large-scale disturbances and an unstable security situation" as well as "grave acts of violence". Others have targeted the country's religious establishment.