Diallyl Trisulfide Activates NADPH Oxidase By Src-Gab1-Shp2 Signaling In Human Promyelocytic HL-60 Cells

Objective: To explore the mechanism of NADPH oxidase activating by diallyl trisulfide (diallyl trisulfide, DATS) in human promyelocytic leukemia HL-60 cells.Methods: HL-60 cells were either treated with the concentration of 150μM of DATS for 0, 5, 15, 30, 60 and 180 min, or pre-incubated with a special Src kinase inhibitor PP2 for 30 min followed by DATS treatment for the same duration. The phosphorylation of Src kinase, Gab1 and Shp2 was detected by Western blot. NADPH oxidase activity was analyzed by NBT reduction test in HL-60 cells.Results: DATS markedly increases the phosphorylation of Src kinase, maximum at 5 and 15 min (P<0.05), meanwhile DATS activates NADPH oxidase. Src kinase inhibitor PP2 significantly decreases the phosphorylation of Src kinase and inhibites NADPH oxidase activity (P<0.05). DATS markedly induces the phosphorylation of Gab1 and Shp2(P<0.05), maximum from 15 to 30 min for Gab1, and at 15 and 180 min for Shp2 . PP2 significantly attenuates the phosphorylation of Gab1 and Shp2(P<0.05).Conclusion: DATS activates Src-Gab1-Shp2 signaling, which is involved in the activation of NADPH oxidase induced by DATS in HL-60 cells.