The invention discloses a preparation method of a material with nitric oxide (NO) catalytic activity. The preparation method is implemented through the steps that a selenium-containing compound with nitric oxide catalytic activity, a sulfur-containing compound, a soluble copper salt, a compound with an o-phenol structure, a flavonoid compound, and a flavonol compound or a flavanone compound are mixed in a buffer solution and then polymerized. The material with NO catalytic activity not only can be applied to the surface modification of materials which are different in material, geometrical shape and topological structure, and also can be used as a filling material for a controlled release system. Double selenium bonds, double sulfur bonds, copper ions and phenolic hydroxyl groups, which are contained in the prepared material with nitric oxide (NO) catalytic activity, have an excellent free radical removal function; the selenium bonds, the sulfur bonds and the chelate copper ions, which are contained in the material, also have a reduced glutathione (GSH) response function; in addition, the copper ions containing in the material also have an antibacterial function; and the material, besides being used for catalyzing NO release, also can be applied to all related fields of free radical removal and GSH response function.