Organizers said the Rotunda rally was about "visibility, resilience, and solidarity."
The event took place just hours before the Kentucky House voted to undo a ban on so-called conversion therapy and prohibit the state's Medicaid program from covering gender transition treatment.
During the rally, Lexington city council member and trans activist Emma Curtis hinted at a possible run for state office.
"In January of 2027, when the legislature gavels in, my promise to you is that we are finally going to have trans representation on the floor of our General Assembly, taking the Oath of Office, ready to hold the line and fight for all of us in Kentucky," Curtis told the audience.
Curtis is a newcomer to Lexington's Urban County Council, having defeated fourth district incumbent Brenda Monarrez in the 2024 elections.