Herein, we prepared CoO nanozymes with three types of enzyme catalytic activities for the first time, which have SOD-like, CAT-like, and POD-like catalytic activities. This is the first study to report the preparation of CoO nanoparticles with three types of enzyme catalytic activities by the one-pot method. By modifying the surface of CoO nanozymes with a carboxyl group, its biocompatibility enhanced, so it can be used in the field of life sciences. In vitro cytotoxicity and anti-HO-induced ROS experiments proved that CoO nanozymes can protect HaCaT cells against ROS and cytotoxicity induced by HO. In addition, an atopic dermatitis (AD) mouse model was established by topical application of MC903, which verified the anti-inflammatory effect of CoO nanozymes on the AD mouse model. Traditional drugs for the treatment of AD, such as dexamethasone, have significant side-effects. The side-effects include skin burns, telangiectasias, and even serious drug dependence. CoO nano-enzymes have a low cytotoxicity and its multiple enzyme-like catalytic activities can effectively protect cells and tissues in ROS environments, which proves that CoO nano-enzymes have high application potential in the field of anti-inflammation.