Diallyl Trisulfide Mediated The Dephosphorylation Of PI3K-Akt-mTOR And Phosphorylation Of P38 Kinase Via Reactive Oxygen Species In HL-60 Cells

Objective: To explore diallyl trisulfide on the phosphorlation of PI3K-Akt-mTOR and p38 kinase , and elucidate the role of reactive oxygen species in the phosphorlation of PI3K-Akt-mTOR and p38 kinase in HL-60 cells.Method: HL-60 cells were either treated with various doses of DATS alone for 0, 1, 3, 6, 12 hours, or150μM DATS combination with NADPH oxidase specife inhibitor apocynin, antioxidant N-acetyl-L-cysteine (NAC), PI3K specife inhibitor LY294002, and p38 specife inhibitor SB203580 for 3 hours, respectively. The intracellular ROS level was detected by flow cytometry. The phosphrylation level of PI3K,Akt,mTOR,p38 was detected by western blot.Results: DATS significantly increased the intracellular ROS level in HL-60 cells. The fluorescence densities of ROS increased in the dose-and time-dependent maner during the time from 1 to 3h, and reached at maximuam(89.7±3.67) when HL-60 cells were incubated with DATS of 150μM for 3 hours, and and then maintain a higher level.DATS induced the dephosphrylation of PI3K, Akt and mTOR, and p38 phosphrylation in a concentration and time-dependent manner when HL-60 cells were treated with the concentration of DATS from 50μM to 200μM, or with 150μM DATS from 1 to 12h. NADPH oxidase specife inhibitor apocynin and antioxidant N-acetyl-L-cysteine promoted the phosphrylation of PI3K and Akt, but decreased p38 phosphrylation. PI3K specific inhibitor LY294002 down-regulated the phosphrylation level of PI3K and Akt. The p38 specific inhibitor SB203580 blocked p38 phosphrylation.Conclusion: DATS induce ROS generation and the dephosphorlation of PI3K-Akt-mTOR and p38 phosphorlation in HL-60 cell. ROS is an important mediator in the phosphorlation of PI3K-Akt-mTOR and p38 kinase.