The ultraviolet-B (UV-B) radiation can affect gene expression of plant in growth and development. Winter wheat (Triticum aestivum), as a kind of economic crop cultured in the Northern China, is also damaged by present-day ultraviolet-B (UV-B) radiation. It was reported that He-Ne lasers could alleviate cell damage caused by UV-B radiation in plants. To get the proteomic changes of wheat alleviated by He-Ne lasers, we studied protein expression profiles of winter wheat (Jin Mai NO.79) leaves by running 2-DE (two-dimensional gel electrophoresis), which allowed the identification of some significantly different gel spots. The spots were further verified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry, in which they were confirmed to be winter wheat proteins. The results showed: 1) the proteomic changes between the ultraviolet-B radiation group and normal light group were significantly different on sixth day; 2) The expression of some proteins such as catalase, DNA polymerase and reduced glutathione declines at the ultraviolet-B radiation group and increases at the UV-B + He-Ne laser group. This indicates UV-B could regulate genes encoding proteins in Winter wheat leaves and affect the growth of the Winter wheat. He-Ne lasers treatment could significantly alleviate UV-B-induced damage in wheat.