Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods...Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods The endothelial cells were scratched from trypsined canine aorta endothelium. HSYA was added to the cells at final concentrations of 1 × 10^-3, 1 × 10^-4 and 1 × 10^-5 mol· L^-1, respectively. VEGF （2.6 × 10^-7 mol· L^-1 ）-treated cells were used as the positive control. The proliferative effect of HSYA on VEC was determined at 48, 72, 96, and 120 h in normoxic culture by MTI＂ assay. Similarly, the proliferation of VEC was determined at 12, 24, 48, and 72 h in hypoxic culture by MTF assay. The effects of HSYA on VEC proliferation and VEGF secretion were investigated by MTr and ELISA assays at the presence of the antibodies to VEGF and VEGF receptors. Results Pretreatment with HSYA at concentrations of 1 × 10^-3 and 1 × 10^-4 mol· L^-1 enhanced VEC proliferation in normoxic culture. The most significant enhancing effect of HSYA on VEC proliferation was achieved at 24, 48, and 72 h in hypoxic culture in concentration-dependent and time-dependent manner. HSYA at 1 × 10^-3 mol·L^-1 showed a potency similar to VEGF at 2.6 × 10^-7 mol·L^-1 . Pretreatment with the antibodies of Flt-1, KDR or VEGF blocked the proliferative effect of HSYA with similar potencies. Antibodies of Fit-1 or VEGF antagonized the promoting effect of HSYA on VEGF secretion. Conclusion HSYA promotes VEC proliferation either in normoxic or hypoxic culture, especially in the latter condition. This effect of HSYA is at least partly mediated by VEGF and VEGF receptor.展开更多
目的:本实验探讨血管内皮生长因子165(VEGF165)对体外培养的胃癌细胞株BGC-823凋亡的影响和机制.方法:将BGC-823细胞分为对照组、感染复数(MOI=20)病毒Ad-GFP的Ad-GFP组,重组腺病毒Ad-VEGF165转染的Ad-VEGF165组.应用流式细胞仪检测细...目的:本实验探讨血管内皮生长因子165(VEGF165)对体外培养的胃癌细胞株BGC-823凋亡的影响和机制.方法:将BGC-823细胞分为对照组、感染复数(MOI=20)病毒Ad-GFP的Ad-GFP组,重组腺病毒Ad-VEGF165转染的Ad-VEGF165组.应用流式细胞仪检测细胞凋亡的百分率,RT-PCR方法检测凋亡抑制基因Bcl-2mRNA的表达,免疫细胞化学方法检测Bcl-2蛋白的表达情况.结果:流式细胞仪测定显示Ad-VEGF165组的细胞凋亡率明显低于Ad-GFP组和对照组(4.6%±0.31% vs 8.37%±1.06%.7.73%±0.86%,P<0.01);RT-PCR和细胞免疫化学结果显示VEGF165转染BGC-823细胞后促进了细胞Bcl-2mRNA和蛋白的表达.Ad-VEGF165组Bcl-2mRNA和蛋白均高于对照组和Ad-GFP组(Bcl-2mRNA:0.761±0.05 vs 0.363±0.12.0.356±0.08;Bcl-2蛋白:1.010±0.08 vs 0.865±0.07,0.901±0.05;P<0.01).结论:VEGF165通过上调凋亡抑制基因Bcl-2及其蛋白的表达,来抑制血浆饥饿诱导的胃癌细胞的凋亡.展开更多
目的观察土茯苓提取物落新妇苷对永生化人角质形成细胞Ha Ca T增殖、凋亡的影响,并探讨其机制。方法收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷、空白对照组加DMEM培养液,培养24 h后,采用MTT法测算细胞增殖抑制率,流式...目的观察土茯苓提取物落新妇苷对永生化人角质形成细胞Ha Ca T增殖、凋亡的影响,并探讨其机制。方法收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷、空白对照组加DMEM培养液,培养24 h后,采用MTT法测算细胞增殖抑制率,流式细胞仪检测细胞早期凋亡率;收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷+20 ng/m L INF-γ、模型对照组加20 ng/m L INF-γ、空白对照组加DMEM培养液培养24 h,采用实时荧光定量PCR法检测细胞中的核转录因子κB(NF-κB)p65、骨桥蛋白(OPN)、血管内皮生长因子(VEGF)mRNA。结果落新妇苷在25、50、100、200、400μg/m L时,Ha Ca T细胞增殖抑制率分别为3.06%、26.53%、40.82%、60.20%及61.22%。落新妇苷在50、100、200μg/m L时,观察组Ha Ca T细胞早期凋亡率分别为3.26%、8.24%和15.38%;与空白对照组早期凋亡率(1.33%)比较,P均<0.05。与空白对照组比较,模型对照组NF-κB p65、OPN、VEGF mRNA表达增加(P均<0.01);与模型对照组比较,观察组随落新妇苷浓度增加,NF-κB p65、OPN、VEGF mRNA表达逐渐降低(P均<0.05)。结论土茯苓提取物落新妇苷能抑制Ha Ca T细胞增殖、诱导细胞凋亡,其机制可能与制抑细胞NF-κB p65、OPN、VEGF mRNA表达有关。展开更多
文摘Aim To study the proliferative effeet of hydroxysaftlor yellow A （HSYA） on cultured canine aortic endothelial cell （VEC） in normoxic （21% O2 ） or hypoxic （10% O2 ） culture and the underlying mechanism. Methods The endothelial cells were scratched from trypsined canine aorta endothelium. HSYA was added to the cells at final concentrations of 1 × 10^-3, 1 × 10^-4 and 1 × 10^-5 mol· L^-1, respectively. VEGF （2.6 × 10^-7 mol· L^-1 ）-treated cells were used as the positive control. The proliferative effect of HSYA on VEC was determined at 48, 72, 96, and 120 h in normoxic culture by MTI＂ assay. Similarly, the proliferation of VEC was determined at 12, 24, 48, and 72 h in hypoxic culture by MTF assay. The effects of HSYA on VEC proliferation and VEGF secretion were investigated by MTr and ELISA assays at the presence of the antibodies to VEGF and VEGF receptors. Results Pretreatment with HSYA at concentrations of 1 × 10^-3 and 1 × 10^-4 mol· L^-1 enhanced VEC proliferation in normoxic culture. The most significant enhancing effect of HSYA on VEC proliferation was achieved at 24, 48, and 72 h in hypoxic culture in concentration-dependent and time-dependent manner. HSYA at 1 × 10^-3 mol·L^-1 showed a potency similar to VEGF at 2.6 × 10^-7 mol·L^-1 . Pretreatment with the antibodies of Flt-1, KDR or VEGF blocked the proliferative effect of HSYA with similar potencies. Antibodies of Fit-1 or VEGF antagonized the promoting effect of HSYA on VEGF secretion. Conclusion HSYA promotes VEC proliferation either in normoxic or hypoxic culture, especially in the latter condition. This effect of HSYA is at least partly mediated by VEGF and VEGF receptor.
文摘目的:本实验探讨血管内皮生长因子165(VEGF165)对体外培养的胃癌细胞株BGC-823凋亡的影响和机制.方法:将BGC-823细胞分为对照组、感染复数(MOI=20)病毒Ad-GFP的Ad-GFP组,重组腺病毒Ad-VEGF165转染的Ad-VEGF165组.应用流式细胞仪检测细胞凋亡的百分率,RT-PCR方法检测凋亡抑制基因Bcl-2mRNA的表达,免疫细胞化学方法检测Bcl-2蛋白的表达情况.结果:流式细胞仪测定显示Ad-VEGF165组的细胞凋亡率明显低于Ad-GFP组和对照组(4.6%±0.31% vs 8.37%±1.06%.7.73%±0.86%,P<0.01);RT-PCR和细胞免疫化学结果显示VEGF165转染BGC-823细胞后促进了细胞Bcl-2mRNA和蛋白的表达.Ad-VEGF165组Bcl-2mRNA和蛋白均高于对照组和Ad-GFP组(Bcl-2mRNA:0.761±0.05 vs 0.363±0.12.0.356±0.08;Bcl-2蛋白:1.010±0.08 vs 0.865±0.07,0.901±0.05;P<0.01).结论:VEGF165通过上调凋亡抑制基因Bcl-2及其蛋白的表达,来抑制血浆饥饿诱导的胃癌细胞的凋亡.
文摘目的观察土茯苓提取物落新妇苷对永生化人角质形成细胞Ha Ca T增殖、凋亡的影响,并探讨其机制。方法收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷、空白对照组加DMEM培养液,培养24 h后,采用MTT法测算细胞增殖抑制率,流式细胞仪检测细胞早期凋亡率;收集对数生长期Ha Ca T细胞,观察组分别加入不同浓度落新妇苷+20 ng/m L INF-γ、模型对照组加20 ng/m L INF-γ、空白对照组加DMEM培养液培养24 h,采用实时荧光定量PCR法检测细胞中的核转录因子κB(NF-κB)p65、骨桥蛋白(OPN)、血管内皮生长因子(VEGF)mRNA。结果落新妇苷在25、50、100、200、400μg/m L时,Ha Ca T细胞增殖抑制率分别为3.06%、26.53%、40.82%、60.20%及61.22%。落新妇苷在50、100、200μg/m L时,观察组Ha Ca T细胞早期凋亡率分别为3.26%、8.24%和15.38%;与空白对照组早期凋亡率(1.33%)比较,P均<0.05。与空白对照组比较,模型对照组NF-κB p65、OPN、VEGF mRNA表达增加(P均<0.01);与模型对照组比较,观察组随落新妇苷浓度增加,NF-κB p65、OPN、VEGF mRNA表达逐渐降低(P均<0.05)。结论土茯苓提取物落新妇苷能抑制Ha Ca T细胞增殖、诱导细胞凋亡,其机制可能与制抑细胞NF-κB p65、OPN、VEGF mRNA表达有关。
