Effect of TSG and PDTC on Apoptosis of HUVECs Induced by H2O2

Abstract:

Objective: To study the anti-apoptotic effect and mechanism of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) combined with pyrrolidine dithiocar-bamate (PDTC) on the apoptosis of human umbilical vein endothelial cells (HUVECs) induced by H₂O₂. Method: HUVECs were divided into five groups: control group, H₂O₂ group (300 μmol·L^-1), TSG group (10 μmol·L^-1 TSG + 300 μmol·L^-1 H₂O₂), TSG and PDTC combination group (75 μmol·L^-1 PDTC + 10 μmol·L^-1 TSG + 300 μmol·L^-1 H₂O₂ group), PDTC group (75 μmol·L^-1 PDTC +300 μmol·L^-1 H₂O₂). The viability of HUVECs was detected by MTT assay. The morphology of apoptosis was observed by Hoechst 33258 staining and the rate of apoptosis was determined by flow cytometry. The protein expressions of NF-κB, Caspase-3 were detected by Western-blot. Result: Compared with control group, H₂O₂ significantly increased the apoptosis, reduced the cell viability, and enhanced the expressions of NF-κB and Caspase-3 (P<0.01). The proliferation rate increased significantly (P<0.01) while the apoptosis rate decreased obviously in TSG or PDTC pretreatment group, and the expression of NF-κB and Caspase-3 down regulated (P<0.05) compared with H₂O₂ group (P<0.01). Combination of PDTC and TSG raised the cell viability and attenuated the cell apoptosis rates than pretreatment of TSG or PDTC alone, and the expression of NF-κB and Caspase-3 decreased significantly (P<0.01). Conclusion: Both PDTC and TSG pretreatment could inhibit HUVECs apoptosis induced by H₂O₂, and the combination of PDTC and TSG can lead to the synergistic effect. The mechanism was associated with the expressions of NF-κB and Caspase-3.