"Ukrainian authorities have recently been pursuing policies that cannot be seen as anything other than unfriendly towards Russia," the foreign ministry said in a toughly-worded statement.
Western officials have expressed concern that Ukraine`s large ethnic Russian population could leave it exposed to intervention from Moscow after its war in the Georgian breakaway region of South Ossetia.
Russia said it was defending Russian nationals when it sent in troops to halt a Georgian offensive on South Ossetia.
Ukrainian President Viktor Yushchenko last month raised tensions by imposing restrictions on the Russian fleet, requiring ships to seek permission at least 72 hours prior to crossing the Ukrainian border.
The announcement, which came after the Black Sea fleet took part in armed conflict with Georgia, also called for talks on the future of the fleet`s base in the southern port of Sevastopol.
The Russian foreign ministry statement said the fleet -- based on the predominantly Russian-speaking Crimean peninsula -- was "a stabilising factor both for relations between Russia and Ukraine and in the context of regional stability."
"The rights of the Russian-speaking population of Ukraine are being abused and there is a policy targeted at excluding the Russian language from the public life of the country," the statement said.
Russia also accused Ukraine of siding with the pro-West government in Georgia, saying: "We have not heard words of pity or compassion on the death of civilians in Tskhinvali and of Russian peacekeepers.
"On the contrary, the Ukrainian president has tried to blame Russia for the bloodshed," it said, adding that Ukraine bore responsibility for casualties because it had supplied Georgia with arms.
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