Anti-radiation Activity Screening Of Phenylethanoid Glycosides And Its Mechanism Research

Objective:1.To study the radiation protection activity in vitro of six kinds of phenylethanoid glycosides on X-ray radiation damaged Human Skin Fibroblasts(HSF),screen out the one of the strongest and determine its optimal dose in vitro.2.Using the selected phenylethanoid glycoside to investigate its influence on cell apoptosis,cell cycle and reactive oxygen species(ROS)level in radiation damaged HSF.3.To discuss the inhibition of the selected phenylethanoid glycoside on abnormal expression of Bcl-2?Bax?proaspase-3?ERK?JNK?p-ERK and p-JNK in radiation damaged HSF.Methods:1.2 Gy?4 Gy?8 Gy?16 Gy?32 Gy one-time X-ray exposure were used respectively to screen and establish the model of radiation damaged HSF,and the MTT assay were used to detect and compared the cell's proliferation activity after treated by six kinds of phenylethanoid glycosides,then screened out the most strongest one and determined its optimal dose.2.Selected the optimal dose of the one which has the strongest radiation protective effect in six to intervene preventively on HSF 2 h before 16 Gy X-ray irradiated,using Annexin V-FITC/PI kit,Propidium Iodide kit and DCFH-DA kit to investigate the effect of phenylethanoid glycosides on cell apoptosis,cell cycle and the production of ROS.3.Western blot assay were used to detect the impact of selected phenylethanoid glycoside on abnormal protein expression of Bcl-2?Bax?proaspase-3?ERK?JNK?p-ERK and p-JNK in radiation damaged HSF.Result:1.16 Gy one-time X-ray irradiation can inhibit proliferation activity of HSF significantly(P<0.01).Six kinds of phenylethanoid glycosides have varying degrees of anti-radiation effect(P<0.01),in which the effect of acteoside is preferably,its optimal dosage is 50 ?g/ml.2.Compared with model group,50?g/ml acteoside can reduce cell apoptosis,the cell cycle of G0/G1 phase and the production of intracellular ROS in HSF significantly,and its inhibitory effects of cell apoptosis and ROS generation are more significant than 50 ?g/ml paeoniflorin,but its regulation function of cell cycle is weaker than 50 ?g/ml paeoniflorin.3.Compared with model group,50 ?g/ml acteoside can effectively inhibit the radiation-induced Bax protein expression increased and Bcl-2?procaspase-3 protein expression decreased,at the same time,it can reduce the phosphorylation of ERK and JNK in radiation damaged HSF to reduce the rate of cell apoptosis.Conclusion:phenylethanoid glycosides,especially 50?g/ml acteoside have significant effect to protect radiation damage.