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投稿时间：2020-01-15
投稿时间：2020-01-15
中文摘要: 该研究探讨草苁蓉多糖（Boschniakia rossica polysaccharides，BRPS）对氧化损伤致人脐静脉血管内皮细胞（human umbilical vein endothelial cells，HUVEC）凋亡相关蛋白表达的影响。以叔丁基过氧化氢（tert-butyl hydroperoxide，t-BHP）损伤HUVEC建立体外凋亡模型，分为正常组、t-BHP组（损伤组）、BRPS组，分别应用细胞增殖毒性检测试剂盒（cell counting kit-8，CCK-8）法和流式细胞术检测细胞存活率和细胞凋亡率，用Western blotting技术检测凋亡相关蛋白胱天蛋白酶原（pro-cysteine aspastic acid-specific protease，Procaspase）、聚腺苷二磷酸核糖聚合酶[poly（ADP-ribose）polymerase，PARP]、Survivin、p53、核转录因子-кB（nuclear factor-кB，NF-кB）信号通路蛋白以及丝裂原活化蛋白激酶（mitogen activated protein kinase，MAPK）信号通路蛋白活化水平。结果显示，与正常组比较，损伤组HUVEC细胞活力降低50.6%，细胞凋亡率升高41.0%；与损伤组比较，BRPS组HUVEC细胞活力升高16.4%，细胞凋亡率降低了22.2%。此外，t-BHP降低HUVEC细胞的Procaspase-3、Procaspase-8、Procaspase-9、PARP和Survivin蛋白水平，升高Cleaved-PARP、p53、NF-κB、磷酸化细胞外信号调节蛋白激酶（phosphorylated extracellular signalregulated kinase，p-ERK）、磷酸化 c-Jun N 末端激酶（phosphorylated c-Jun N-terminal kinase，p-JNK）、磷酸化 p38丝裂原活化蛋白激酶（phosphorylated p38 mitogen-activated protein kinase，p-p38）蛋白水平。而BRPS升高Procaspase-3、Procaspase-8、Procaspase-9、PARP 和 Survivin 蛋白水平，降低 Cleaved-PARP、p53、NF-кB、p-JNK、p-p38 蛋白水平。研究结果表明，BRPS对t-BHP引起HUVEC凋亡具有抑制作用，其机制可能与JNK、p38及NF-κB调控有关。
中文关键词: 草苁蓉 多糖 叔丁基过氧化氢 人脐静脉血管内皮细胞 细胞凋亡
Abstract:The effect of Boschniakia rossica polysaccharides （BRPS）on expression of apoptosis-related proteins of human umbilical vein endothelial cells（HUVEC）induced by oxidative damage was investigated.HUVEC were treated with tert-butyl hydroperoxide （t-BHP）to establish apoptosis model in vitro，and then assigned into the normal，model and BRPS groups.Cell viability and apoptosis rate of HUVEC were detected by CCK-8 and flow cytometry methods，respectively.The western blot analysis was used to detect the expression of the apoptosis-associated proteins，such as pro-cysteine aspastic acid-specific protease （Procaspase），[poly（ADP-ribose）polymerase] （PARP），survivin，p53，nuclear factor-кB （NF-кB）and mitogen activated protein kinase（MAPK）proteins.Results showed that cell viability of the model group was decresed by 50.6%and apoptosis rate was increased by 41.0%as compared with the normal group，while cell viability of BRPS group was increased by 16.4%and apoptosis rate was decreased by 22.2%as compared with the model group.In addition，t-BHP reduced the level of procaspase-3，procaspase-8，procaspase-9，PARP and survivin proteins，and elevated the expression of cleaved-PARP，p53，NF-кB，phosphorylated extracellular signal-regu－lated kinase（p-ERK），phosphorylated c-Jun N-terminal kinase（p-JNK），phosphorylated p38 mitogen-activated protein kinase （p-p38 proteins.However，BRPS up-regulated the protein expression of procaspase-3，procaspase-8，procaspase-9，PARP and survivin，and down-regulated the protein expression of cleaved-PARP、p53、NF-кB、p-JNK，p-p38.In conclusion，BRPS inhibit the apoptosis of HUVEC induced by t-BHP，and the mechanism may be related to the regulation of JNK and p38 and NF-кB.
keywords: Boschniakia rossica polysaccharides tert-butyl hydroperoxide human umbilical vein endothelial cells apoptosis
文章编号：202022027 中图分类号： 文献标志码：
基金项目:国家自然科学基金（81760659、81360651）
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