异鼠李素通过减轻氧化应激延缓D-半乳糖诱导的人脐静脉内皮细胞衰老

doi:10.3969/j.issn.1004-616x.2021.02.003

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (2): 95-100.doi: 10.3969/j.issn.1004-616x.2021.02.003

异鼠李素通过减轻氧化应激延缓D-半乳糖诱导的人脐静脉内皮细胞衰老

蒋杉¹, 杜凤¹, 康秉文², 殷草草², 周健¹, 石莹², 王玥², 周耿瑶¹, 秦绪军¹

1. 空军军医大学军事预防医学系, 军队健康教育与管理教研室, 陕西西安 710032;
2. 陕西中医药大学公共卫生学院, 陕西咸阳 712046

收稿日期:2020-11-06 修回日期:2020-12-25 出版日期:2021-03-30 发布日期:2021-04-12
通讯作者: 秦绪军,E-mail:qinxujun@hotmail.com E-mail:qinxujun@hotmail.com
作者简介:蒋杉,E-mail:490731543@qq.com。
基金资助:
国家自然科学基金（31670863）；陕西省自然科学基金（2016JZ027）

Attenuation of D-galactose-induced senescence of human umbilical vein endothelial cells by isorhamnetin

JIANG Shan¹, DU Feng¹, KANG Bingwen², YIN Caocao², ZHOU Jian¹, SHI Ying², WANG Yue², ZHOU Gengyao¹, QIN Xujun¹

1. Department of Military Health Education and Management, School of Preventive Medicine, the Air Force Medical University, Xi'an 710032;
2. School of Public Health, Shaanxi University of Chinese Medicine, Xianyang 712046, Shaanxi, China

Received:2020-11-06 Revised:2020-12-25 Online:2021-03-30 Published:2021-04-12

摘要/Abstract

摘要： 目的：探讨沙棘中的重要成分异鼠李素（ISO）对细胞衰老的影响及机制。方法：采用D-半乳糖（D-Gal）诱导细胞衰老模型，取20～23代的人脐静脉内皮细胞（HUVEC）分为4组：对照组、D-Gal组、D-Gal+ISO组（5 μmol/L）、D-Gal+ISO组（10 μmol/L）。其中D-Gal浓度为10 g/L，干预时间为72 h。采用CCK-8试剂盒检测各组细胞活力；衰老相关β-半乳糖苷酶（SA-β-Gal）染色试剂盒检测细胞衰老状态；各相关试剂盒检测细胞总活性氧（ROS）水平、丙二醛（MDA）含量、总超氧化物歧化酶（SOD）活性和过氧化氢酶（CAT）活性。Western blot法检测衰老标志蛋白p21、p27和核因子NF-E2相关因子2（Nrf2）及其下游蛋白表达水平。结果：ISO浓度≤10 μmol/L时对HUVEC无明显毒性作用；与对照组比较，D-Gal能够诱导细胞衰老指标SA-β-Gal活性增加（P<0.05）；提高p21、p27表达水平，提高细胞总ROS水平和MDA含量，降低总SOD和CAT活性，降低Nrf2及下游蛋白表达水平（P<0.05）。与D-Gal组比较，ISO能够降低细胞SA-β-Gal活性，抑制p21、p27表达，降低细胞总ROS水平和MDA含量，提高总SOD和CAT活性，增强Nrf2及下游蛋白表达（P<0.05）。结论：ISO可以通过减轻氧化应激，延缓D-Gal诱导的HUVEC细胞衰老，在此过程中激活Nrf2通路可能是其发挥抗衰老作用的重要机制。

关键词: 异鼠李素, 人脐静脉内皮细胞, 氧化应激, 细胞衰老

Abstract: OBJECTIVE: To investigate effects and mechanisms of isorhamnetin (ISO),an important component of sea-buckthorn,on aging. METHODS: In this study,the model of D-galactose (D-Gal)-induced aging was employed. Human umbilical vein epithelial cells (HUVEC) in their 20th-23th generations were divided into 4 groups:control,D-Gal,D-Gal+5 μmol/L ISO and D-Gal+10 μmol/L ISO groups. The D-Gal concentration was 10 g/L and the intervention time was 72 hours. CCK-8 kit was used to detect cell viability in each group. Senescence was determined by the Senescence-associated β-Galactosidase (SA-β-Gal) staining kit. Levels of total reactive oxygen species (ROS),malondialdehyde (MDA),superoxide dismutase (SOD) activities and catalase (CAT) activities were detected by their respective kits. Western Blot was used to detect expression levels of aging markers,p21 and p27,as well as nuclear factor,erythroid-2 related factor 2 (Nrf2),and several downstream molecules. RESULTS: Isorhamnetin with concentration ≤ 10 μmol/L had no obvious toxic effects on HUVEC. Compared with the control group,D-Gal treatment increased the activity of SA-β-Gal,the protein levels of p21 and p27,and the total ROS level and MDA content. At the same time,D-Gal treatment inhibited the activities of total SOD and CAT,the protein levels of Nrf2 and the downstream molecules (P<0.05). Compared with the D-Gal group,isorhamnetin treatment effectively inhibited the activity of SA-β-Gal,the protein levels of p21 and p27,and the total ROS level and MDA content. Moreover,isorhamnetin treatment improved the activities of total SOD and CAT,and enhanced the protein levels of Nrf2 and its downstream molecules. CONCLUSION: Isorhamnetin attenuated D-Gal-induced HUVEC senescence by reducing oxidative stress and activation of Nrf2 pathway might be an important mechanism.

Key words: isorhamnetin, HUVEC, oxidative stress, senescence

中图分类号:

R282.71

蒋杉, 杜凤, 康秉文, 殷草草, 周健, 石莹, 王玥, 周耿瑶, 秦绪军. 异鼠李素通过减轻氧化应激延缓D-半乳糖诱导的人脐静脉内皮细胞衰老[J]. 癌变·畸变·突变, 2021, 33(2): 95-100.

JIANG Shan, DU Feng, KANG Bingwen, YIN Caocao, ZHOU Jian, SHI Ying, WANG Yue, ZHOU Gengyao, QIN Xujun. Attenuation of D-galactose-induced senescence of human umbilical vein endothelial cells by isorhamnetin[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(2): 95-100.

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异鼠李素通过减轻氧化应激延缓D-半乳糖诱导的人脐静脉内皮细胞衰老

Attenuation of D-galactose-induced senescence of human umbilical vein endothelial cells by isorhamnetin

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