|Keratinocytes is an important role to protecting the skin from the external environment for a variety of stimuli to the skin, and the inner material and the water is the key to defend the skin from being lost out. However, it is still largely unknown about effective protectors against ultraviolet B (UVB) or oxidative stress in human keratinocyte, HaCaT cells. Therefore, the purpose of this study was in exploring the ingredients to prepare a Paeonia suffruticosa extracts, we were evaluated for the cytoprotective effect on the oxidative damage induced by UVB and Hydrogen peroxide(H2O2). P. suffruticosa extracts showed the large amount of polyphenol content and total flavonoid content. In addition, DPPH scavenging, total antioxidant capacity and superoxide scavenging activity also showed a superior effect by P. suffruticosa extracts more than the positive control. Furthermore, P. suffruticosa extracts also exhibited an effect of HaCaT cells protection significantly against the UVB or H2O2 toxicity. Equivalent to P. suffruticosa extracts showed the effect of significantly inhibiting the DNA segment and pro-caspase 9 degradation on the UVB or H2O2-induced HaCaT cells. Based on the result of this comprehensive P. suffruticosa extracts have excellent antioxidant effects, and it also have protected the keratinocytes against the UVB or H2O2 HaCaT cells toxicity. Therefore, these results suggest that P. suffruticosa extracts can be used the development potential as a cosmetic material. The further study is necessary accordingly to find a standardization of the active ingredient.