This study was conducted to explore the effect of CuO nanoparticles (NPs) on the cell membrane permeability and its exclusion from cells. Human A549 lung cells were exposed to 5mg/L and 15mg/L CuO NPs. Cell membrane permeability was evaluated in 2h and 4h. After 4 hours exposure, the membrane was damaged. Exclusion of copper from cells after 24h exposure with 5mg/L and 15 mg/L CuO NPs are time and dose dependent. And the cell viability was resumed gradually. It is concluded that CuO NPs could induce cytotoxicity, and destroy the membrane integrity. One detoxify mechanism was the exclusion of excessive copper from cells themselves.