Jassem al-Khorafi told reporters in parliament on Saturday that Kuwait did not accept any form of interference in its sovereignty.
"As we respect the views of others and we do not interfere in their internal affairs, I hope and insist that others should not interfere in our affairs and should not move their conflicts with other countries to Kuwait," the Kuwaiti parliamentary speaker said.
The US Ambassador to Kuwait, Richard LeBaron, had blasted Tehran in a lecture at Kuwait's military academy last week, accusing it of supporting terrorism and planning to produce nuclear weapons.
Tehran denies both accusations and says its nuclear programme is for purely peaceful means, which it has the right to pursue.
Washington has repeatedly accused Tehran of seeking to produce nuclear weapons, and LeBaron said "the GCC [Gulf Cooperation Council] should request more than just assurances that Iran's nuclear intent is peaceful."
"It should insist that Iran immediately cease and dismantle its fissile material production effort."
"I hope and insist that others ... should not move their conflicts with other countries to Kuwait"
Kuwait Parliament Speaker
A text of the closed-doors lecture was released by the US embassy earlier this week during a two-day official visit by Iranian Foreign Ministry spokesman Hamid-Reza Asefi to the emirate.
"I blame the Defence Ministry for inviting the US ambassador to deliver such lectures," al-Khorafi said and added that no foreign ambassador should be invited to deliver lectures at state establishments.
"The duties of ambassadors are well-known and do not involve raising issues related to Kuwait's sovereignty," he said.
In the lecture, LeBaron accused Iran of "placing its neighbours, including Kuwait, in harm's way by choosing to pursue nuclear weapons instead of the path of peace".
"The GCC has, within its power, the ability to act together to block this threat," the ambassador said.
The Kuwaiti government, which maintains close diplomatic and military ties with the US, has not publicly criticised the US envoy. But the Defence Ministry said in a brief statement that LeBaron remarks did not reflect its views.