Anti-inflammatory effects of anthocyanins from black soybean seed coat on the keratinocytes and ischemia-reperfusion injury in rat skin flaps

Ischemia‐reperfusion injury is a phenomenon that occurs when tissues are subjected to ischemia for a variable period of time, and then reperfused. Inflammatory reaction has been implicated as one of the most important mechanism of ischemia‐reperfusion injury. The purpose of this study was to evaluate the anti‐inflammatory effects of anthocyanins from black soybean seed coat on keratinocytes in vitro and ischemia‐reperfusion injury in vivo. We investigated the inhibition, by anthocyanins, of the expression of various inflammatory genes associated with ischemia‐reperfusion injury in the tumor necrosis factor‐alpha‐treated (TNF‐α) immortalized epidermal keratinocyte cell line (HaCaT). We also investigated the effects of anthocyanins on the survival of skin flaps after ischemia‐reperfusion injury in the rats. According to Western blot analysis and a luciferase activity assay, anthocyanins inhibited TNF‐α‐induced intercellular adhesion molecule‐1 and cyclooxygenase‐2 (COX‐2) levels through the NF‐κB‐dependent pathway. Administration of anthocyanins (50 and 100 mg/kg) significantly improved the flap area survival in the 10‐hour ischemic model from 62% to 74.5% and 83%, respectively (P = 0.001). The related cytokines in skin flap also changed as the same pattern as in vitro. Our results indicate that anthocyanins from black soybean seed coat had anti‐inflammatory effects on the HaCaT cell line and increase the survival of skin flaps through anti‐inflammatory properties against ischemia‐reperfusion injury. © 2012 Wiley Periodicals, Inc. Microsurgery, 2012.