It called in a statement on "all political actors, and in particular the government, to ensure full respect for human rights and fundamental freedoms and the existing constitution".
And it urged them to "prevent or suspend ongoing initiatives such as the new Constituent Assembly which, instead of fostering reconciliation and peace, foment a climate of tension".
Maduro was due to install the new assembly of his allies today, dismissing an international outcry and opposition protests saying he is burying democracy in his country.
"The Holy See appeals firmly to all of society to avoid all forms of violence and invites, in particular, the security forces to refrain from excessive and disproportionate use of force," the Vatican said.
Forty countries have admonished Maduro for seeing through the creation of the new assembly, with the US hitting him with sanctions this week and US President Donald Trump branding him a dictator.
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