二苯乙烯苷联合PDTC抗H_2O_2诱导的人脐静脉内皮细胞凋亡  

Effect of TSG and PDTC on Apoptosis of HUVECs Induced by H_2O_2

作　　者：龙石银[1] 杨滢[1] 高细强[1] 刘英[1] 乔新惠[1] 张彩平[1] 陈志军[1] 田英[1] 

机构地区：[1]南华大学生物化学与分子生物学教研室,湖南衡阳421001

出　　处：《中国实验方剂学杂志》2013年第8期213-217,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(30800474);湖南省教育厅重点项目(11A104);湖南省科技厅项目(2010SK3037);湖南省研究生创新项目(CX2010B385;CX2011B378)

摘　　要：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 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（P0.01）.The proliferation rate increased significantly（P0.01） while the apoptosis rate decreased obviously in TSG or PDTC pretreatment group,and the expression of NF-κB and Caspase-3 down regulated（P0.05） compared with H2O2 group（P0.01）.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（P0.01）.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.

关 键 词：二苯乙烯苷 人脐静脉内皮细胞 细胞凋亡 核因子-ΚB 

分 类 号：R284.1[医药卫生—中药学] R322.12[医药卫生—中医学]