二苯乙烯苷对AngⅡ诱导人脐静脉内皮细胞衰老的保护作用
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摘要
目的探讨二苯乙烯苷对血管紧张素Ⅱ(AngⅡ)诱导人脐静脉内皮细胞衰老的保护作用。方法体外培养人脐静脉内皮细胞后,AngⅡ诱导细胞衰老模型,MTS法检测细胞存活率,SA-β-gal染色分析细胞衰老程度,qPCR检测miR-34a mRNA表达,Western blot检测p53、PAI-1、SIRT1、E2F1蛋白表达。结果二苯乙烯苷干预后,细胞存活率、SIRT1蛋白表达显著升高(P<0.05),β-gal阳性率及p53、PAI-1蛋白表达显著降低(P<0.05)。同时,该成分对miR-34a、E2F1表达也有明显抑制作用(P<0.05)。结论二苯乙烯苷对AngⅡ诱导人脐静脉内皮细胞衰老具有明显保护作用,其作用机制可能与抑制miR-34a、E2F1表达及促进SIRT1蛋白表达有关。
AIM To explore the protective effects of 2,3,5,4-tetrahydroxystilbene-2-O-β-D-glucoside on AngⅡ-induced human umbilical vein endothelial cellular senescence.METHODS After in vitro cultivation of human umbilical vein endothelial cells, the cellular senescence model was induced by Ang II, MTS assay, SA-β-gal staining, qPCR and Western blot were performed to detect cell viability, analyze cellular senescence degree, and investigate miR-34a mRNA expression, and the protein expressions of p53, PAI-1, SIRT1, E2F1, respectively.RESULTS After 2,3,5,4-tetrahydroxystilbene-2-O-β-D-glucoside intervention, significantly increased cell viability and SIRT1 protein expression(P<0.05), and markedly decreased β-gal positive rate and p53, PAI-1 protein expressions(P<0.05) were found. Meanwhile, this constituent also demonstrated its obvious inhibition on the expressions of miR-34 a and E2F1(P<0.05).CONCLUSION 2,3,5,4-Tetrahydroxystilbene-2-O-β-D-glucoside exhibits obvious protective effects on AngⅡ-induced human umbilical vein endothelial cellular senescence, whose mechanisms may contribute to its inhibition on miR-34 a, E2F1 expressions and promotion on SIRT1 protein expression.
AIM To explore the protective effects of 2,3,5,4-tetrahydroxystilbene-2-O-β-D-glucoside on AngⅡ-induced human umbilical vein endothelial cellular senescence.METHODS After in vitro cultivation of human umbilical vein endothelial cells, the cellular senescence model was induced by Ang II, MTS assay, SA-β-gal staining, qPCR and Western blot were performed to detect cell viability, analyze cellular senescence degree, and investigate miR-34a mRNA expression, and the protein expressions of p53, PAI-1, SIRT1, E2F1, respectively.RESULTS After 2,3,5,4-tetrahydroxystilbene-2-O-β-D-glucoside intervention, significantly increased cell viability and SIRT1 protein expression(P<0.05), and markedly decreased β-gal positive rate and p53, PAI-1 protein expressions(P<0.05) were found. Meanwhile, this constituent also demonstrated its obvious inhibition on the expressions of miR-34 a and E2F1(P<0.05).CONCLUSION 2,3,5,4-Tetrahydroxystilbene-2-O-β-D-glucoside exhibits obvious protective effects on AngⅡ-induced human umbilical vein endothelial cellular senescence, whose mechanisms may contribute to its inhibition on miR-34 a, E2F1 expressions and promotion on SIRT1 protein expression.
引文
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[3] 蔡少艾,赵甘剑,黄尹,等.血管紧张素(1-7)抑制异丙肾上腺素诱导的H9c2心肌细胞凋亡[J].中国动脉硬化杂志,2018,26(5):438-444.
[4] 李亦晗,王跃飞,朱彦.何首乌二苯乙烯苷抗衰老研究进展[J].中国中药杂志,2016,41(2):182-185.
[5] 国家药典委员会.中华人民共和国药典:2015年版[S].北京:中国医药科技出版社,2015:175-177.
[6] 陈冰冰,姜爱玲,张岩.何首乌有效成分二苯乙烯苷的药理活性研究进展[J].中国临床药理学与治疗学,2016,21(6):710-715.
[7] 胡存华,赵立波,王晓敏,等.二苯乙烯苷增强正常血管内皮细胞抗氧化作用研究[J].医药导报,2007,26(2):138-139.
[8] 高王宣,胡英杰,符林春.何首乌二苯乙烯苷的调节血脂作用[J].中国中药杂志,2007,32(4):323-326.
[9] 甘受益,李彩蓉,曾令勇,等.二苯乙烯苷上调SIRT1蛋白防治心肌缺血再灌注损伤[J].湖北科技学院学报(医学版),2014,28(3):188-190.
[10] 陈厚早,刘德培.衰老的基础研究现状与干预策略[J].蚌埠医学院学报,2018,43(10):1261-1265.
[11] 宋云林,谭秋婵,马燕,等.p53-miR-34a-SIRT1反馈环在血管内皮祖细胞复制性衰老过程中的调控作用及机制[J].中国循环杂志,2018,33(5):496-500.
[12] 周丹,刘平,赵德海,等.衰老标记蛋白-30和β-半乳糖苷酶的表达与晶状体上皮细胞凋亡相关性研究[J].哈尔滨医科大学学报,2017,51(5):422-426.
[13] Eren M,Boe A E,Murphy S B,et al.PAI-1-regulated extracellular proteolysis governs senescence and survival in Klotho mice[J].Proc Natl Acad Sci USA,2014,111(19):7090-7095.
[14] Poulsen C R,Watts C A,Murphy J R,et al.Glucocorticoids induce senescence in primary human tenocytes by inhibition of sirtuin 1 and activation of the p53/p21 pathway:in vivo and in vitro evidence[J].Osteoarthr Cartilage,2014,73(7):1405-1413.
[15] Hong E H,Lee S J,Kim J S,et al.Ionizing radiation induces cellular senescence of articular chondrocytes via negative regulation of SIRT1 by p38 kinase[J].J Biol Chem,2010,285(2):1283-1295.
[16] 李杨.细胞转录因子E2F1对下游基因的调控及其生物学功能研究[D].上海:华东师范大学,2010.
[17] Zhang Z,Liu W,Zhao L,et al.Retinoblastoma 1 protects T cell maturation from premature apoptosis by inhibiting E2F1[J].Development,2018,145(1):139-158.
[18] Wang Y,Zhou Y,Xiao L,et al.E2f1 mediates high glucose-induced neuronal death in cultured mouse retinal explants[J].Cell Cycle,2017,16(19):1824-1834.
[19] Badi I,Burba I,Ruggeri C,et al.MicroRNA-34a induces vascular smooth muscle cells senescence by SIRT1 downregulation and promotes the expression of age-associated pro-inflammatory secretory factors[J].J Gerontol A Biol Sci Med Sci,2015,70(11):1304-1311.
[20] Ma W,Xiao G G,Mao J,et al.Dysregulation of the miR-34a-SIRT1 axis inhibits breast cancer stemness[J].Oncotarget,2015,6(12):10432-10444.