The idea of reconciliation emerged in newspapers this week after former reformist president Mohammad Khatami mentioned it in a statement calling on people to join a march marking the 1979 Islamic revolution.
"Some are speaking about the idea of national reconciliation," Khamenei told a public gathering in Tehran, without mentioning any names.
"I think such talks are meaningless. People are united where Islam, Iran and fighting the enemy are concerned.
"Yes, some may have political differences (but) why do you speak of reconciliation? Are people sulking?" he added.
The nation was angry with them "and we will not reconciliate with them," said Khamenei.
He was referring to demonstrations in late 2009 following the election of former president Mahmoud Ahmadinejad.
Authorities accuse protesters who took to the streets on December 27, 2009 of insulting Islam by celebrating on a day of mourning as well as attacking the Basiji militia.
Khamenei said they used the election as an "excuse" while aiming to target the "basis of the revolution".
That year's protests -- which officials call the "sedition" -- were organised after calls by two losing candidates in the elections, Mir Hossein Mousavi and Mahdi Karoubi.
The pair, who alleged the vote was rigged, have been under house arrest since February 2011.
At the time, many reformist figures were imprisoned for "acting against national security" and "propaganda against the regime". Most have since been freed.
Khatami, who is accused of playing an important role in the "sedition", faces strict restrictions.
The media is banned from naming Khatami directly or publishing pictures of him, and he is prevented from attending official ceremonies.
He urged people on February 7 to take part in this year's revolution day march to "neutralise the conspiracies", calling for "national reconciliation".
"I believe that now is the best situation to put in place in the country an atmosphere of national reconciliation," Khatami wrote in his statement.
"People have no doubts in resisting and confronting any threats against the basis of the (Islamic) system," he said.
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