不同浓度雌二醇对过氧化氢诱导的人脐静脉内皮细胞衰老模型的影响

doi:10.13418/j.issn.1001-165x.2019.05.010

摘要/Abstract

摘要： 目的探究不同浓度的17β-雌二醇（17β-Estradiol，17β-E2）对过氧化氢（hydrogen peroxide，H2O2）诱导的人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVECs）衰老模型的影响。方法　构建HUVECs 衰老模型后将细胞分为空白组，衰老组，17β-E2阴性对照组，低浓度、生理浓度及高浓度17β-E2组，CCK-8检测细胞活力；Western blot检测细胞衰老程度相关蛋白p-Rb、Rb、细胞自噬水平相关蛋白P62、LC3B以及衰老相关β半乳糖苷酶（SA-β-Gal）染色。结果　与空白组相比，衰老组p-Rb/Rb比值增加，细胞活力下降，SA-β-Gal染色阳性细胞数目多，与衰老组比较，高浓度和生理浓度17β-E2组p-Rb/Rb比值降低，SA-β-Gal染色阳性细胞数减少，P62表达量减少，LC3-Ⅱ/LC3-I蛋白表达升高，细胞活力增加。结论　17β-E2能通过上调自噬减轻H2O2诱导的HUVECs衰老，且与其浓度有关。

关键词: 自噬, 　血管内皮细胞, 　衰老, 　17β-雌二醇

Abstract: Objective　To investigate the effects of different concentrations of estradiol on senescence model of human umbilical vein endothelial cells (HUVECs) induced by proper concentrations of hydrogen peroxide.　Methods　HUVECs were cultured and the senescence model was successful constructed induced by hydrogen peroxide, then the cultured cells were divided into the following five groups: a normal control, a senescence cells group, a low concentration 17β-E2 group, a physiological concentration 17β-E2 group and a high concentration 17β-E2 group. As results, we performed CCK-8 to detect cell viability, Western blot was conducted to evaluate the expression of the senescence related marker protein (p-Rb and Rb) and autophagocytosis related marker protein (P62 and LC3B), and the β-galactosidase staining also was tested to count the positive cells.　Results　As compared to the normal control, the senescence cells group had a higher protein expression value of p-Rb/Rb, decreased cells viability and more positive cells showed on β-galactosidase staining. On the contrary, in the physiological and high concentration groups, they not only had a lower protein expression value of p-Rb/Rb, increased cells viability, lesser positive cells showed on β-galactosidase staining than the senescence cells group but also had a decreased P62 protein expression and increased expression of LC3-Ⅱ/LC3-I protein which was closely related to autophagocytosis.　Conclusion 17β-E2 with concentration-dependent efficacy can inhibit HUVECs senescence induced by a proper concentrations of hydrogen peroxide through up-regulation of autophagocytosis.

中图分类号:

林捷琪, 阮云军, 杨儒于, 王玉筵, 苏双, 吴赛珠. 不同浓度雌二醇对过氧化氢诱导的人脐静脉内皮细胞衰老模型的影响[J]. 中国临床解剖学杂志, 2019, 37(5): 528-533.

LIN Jie-qi, RUAN Yun-jun, YANG Ru-yu, WANG Yu-yan, SU Shuang, WU Sai-zhu. Investigating the effects of different concentrations of estradiol on senescence model of human umbilical vein endothelial cells induced by hydrogen peroxide[J]. Chinese Journal of Clinical Anatomy, 2019, 37(5): 528-533.

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