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目的:研究二苯乙烯苷(2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside,TSG)联合吡咯烷二硫代氨基甲酸盐(pyrrolidine dithiocarbamate,PDTC)对过氧化氢诱导的人脐静脉内皮细胞凋亡的作用及机制。方法:体外培养人脐静脉内皮细胞,实验分为正常对照组、模型组(300μmol·L-1H2O2)、TSG组(10μmol·L-1TSG+300μmol·L-1H2O2)、联合预处理组(75μmol·L-1PDTC+10μmol·L-1TSG+300μmol·L-1H2O2)、PDTC组(75μmol·L-1PDTC+300μmol·L-1H2O2),采用MTT法检测细胞增殖率,Hoechst 33258染色观察凋亡细胞形态,流式细胞术检测细胞凋亡率,Western blot检测Caspase-3,核因子-κB(NF-κB)蛋白的表达。结果:与正常对照组比较,模型组细胞凋亡率增加,细胞增殖率降低;Caspase-3,NF-κB蛋白表达增加,差异均有显著性(P<0.01)。与模型组相比,经TSG或PDTC预处理后,细胞的增殖率增加,细胞凋亡率降低;Caspase-3,NF-κB蛋白表达显著减少(P<0.01)。联合预处理组与PDTC组或TSG组相比,细胞活力增加,细胞凋亡率下降;Caspase-3,NF-κB蛋白表达进一步减少(P<0.01)。结论:PDTC或TSG预处理可抑制H2O2诱导的人脐静脉内皮细胞凋亡,PDTC可联合TSG协同发挥抗H2O2诱导的内皮细胞凋亡,其机制与抑制NF-κB、Caspase-3的表达有关。
Objective: To investigate the effect of 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) combined with pyrrolidine dithiocarbamate (PDTC) Hydrogen - induced Apoptosis in Human Umbilical Vein Endothelial Cells and Its Mechanism. Methods: Human umbilical vein endothelial cells were cultured in vitro. The experiment was divided into normal control group, model group (300μmol·L-1H2O2), TSG group (10μmol·L-1TSG + 300μmol·L-1H2O2) L-1PDTC + 10μmol·L-1TSG + 300μmol·L-1H2O2) and PDTC group (75μmol·L-1PDTC + 300μmol·L-1H2O2). The cell proliferation rate was detected by MTT assay. The morphology of apoptotic cells was observed by Hoechst 33258 staining. Flow cytometry was used to detect the apoptosis rate. Western blot was used to detect the expression of Caspase-3 and NF-κB. Results: Compared with the normal control group, the apoptosis rate of the model group increased and the cell proliferation rate decreased. The expression of Caspase-3 and NF-κB increased, the difference was significant (P <0.01). Compared with the model group, pretreatment with TSG or PDTC increased the cell proliferation rate and decreased the apoptosis rate; the expression of Caspase-3 and NF-κB significantly decreased (P <0.01). Compared with the PDTC group or the TSG group, the cell viability increased and the apoptosis rate decreased in the combined pretreatment group; the expression of Caspase-3 and NF-κB decreased further (P <0.01). CONCLUSION: Pretreatment with PDTC or TSG can inhibit H2O2-induced apoptosis of human umbilical vein endothelial cells. PDTC can be combined with TSG to exert anti-H2O2-induced endothelial cell apoptosis. The mechanism is related to the inhibition of the expression of NF-κB and Caspase-3.