In order to evaluate the effect of the interface structure on the stability and bioaccessibility of lutein nanostructured lipid carriers (Lutein-NLCs)， whey protein isolate and Tween 80 were applied to design Lutein-NLCs with single-layer， double-layer and complex interface structures. The basic properties， interface adsorption characteristics， bioaccessibility as well as stability of Lutein-NLCs with differen interface structures were investigated. The results showed that the particle sizes of Lutein-NLCs were in the rage of 127.60-180.86 nm. The maximum encapsulation efficiency of Lutein-NLCs could reach 97.78%， and lutein bioaccessibility of Lutein-NLCs with differen interface structures was significantly higher than that of ordinary nanoemulsions (P<0.05). Additionally， lutein bioaccessibility of Lutein-NLCs with single interface structure was significantly higher than that of composite interface and double interface structures (P<0.05). The release rate of lutein in of Lutein-NLCs with three types of interface structures was significantly lower than that of ordinary nanoemulsions. Furthermore， the sustained-release effect of Lutein-NLCs with composite interface and double-interface structures is more significant compared to single-interface Lutein-NLCs. During storage， Lutein-NLCs could increase the lutein retention rate， and the retention rate of Lutein-NLCs with composite interface was the highest， which was 12.06 times that of control group.