The 350 sleek roadsters with a combined horsepower of 190,000 formed a four-kilometre (2.5-mile) long convoy that began the tour from the Sforza Castle in Milan city centre and will end on Saturday at the company's headquarters near Bologna.
Drivers from 29 countries are taking part in the Lamborghini tour, which includes cars owned by the company and as well as aficionados, organizers said.
The biggest representation was from Britain, with 71 cars, followed by Germany, Italy and Switzerland with 30 Lamborghinis each.
Indonesia, Japan, Kuwait, Lebanon, Singapore, South Korea, Tunisia and Ukraine were also among those represented.
"Lamborghini has been the dream and the symbol of Made in Italy for 50 years," said company chief executive Stephan Winkelmann, adding that the tour "reflects the global dimension we have attained".
The tour will cruise through high-end beach resort Forte dei Marmi and Rome, as well as the mediaeval towns of Orvieto and Arezzo.
Lamborghini started out in 1948 as "Lamborghini Trattori", which has since been sold off, while the luxury car division was founded in 1963.
It now has 120 dealerships around the world and is part of the Volkswagen Group.
Dubai police last month announced they had bought a Lamborghini patrol car valued at $550,000 (418,000 euros) to reinforce an image of "luxury and prosperity" in tourist-heavy areas.