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白藜芦醇通过上调人脐静脉内皮细胞自噬减轻过氧化氢诱导的衰老。

Resveratrol attenuates hydrogen peroxide-induced aging through upregulation of autophagy in human umbilical vein endothelial cells.

作者信息

Du Ligen, Chen Enping, Wu Ting, Ruan Yunjun, Wu Saizhu

机构信息

Department of Geriatrics, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China,

Department of Cardiology, The Second People's Hospital of Longgang District, Shenzhen, Guangdong, China.

出版信息

Drug Des Devel Ther. 2019 Feb 22;13:747-755. doi: 10.2147/DDDT.S179894. eCollection 2019.

Abstract

PURPOSE

Resveratrol (RESV; trans-3,5,4'-trihydroxystilbene) has emerged as a potential new therapeutic for age-related atherosclerotic diseases. However, the effect of RESV on cellular aging and its underlying mechanisms remain unknown. Therefore, the aim of this study was to examine whether RESV can delay cellular aging through upregulation of autophagy.

MATERIALS AND METHODS

Human umbilical endothelial vein cells (HUVECs) were divided into four groups: the control group, and the hydrogen peroxide (HO) alone, HO + RESV pretreatment, and HO + 3-methyladenine (3-MA) + RESV pretreatment intervention groups. The cell viability was evaluated by a cell counting kit-8 assay. Superoxide dismutase (SOD) activity and intracellular reactive oxygen species (ROS) levels were tested using commercial kits. Senescence-related β-galactosidase activities were detected by immunohistochemical staining. The expression levels of aging-related and autophagy-related markers, including phosphorylated Rb (p-Rb), LC3, and p62, with or without RESV were measured by Western blotting.

RESULTS

Pretreatment with 10 µM RESV increased the cell viability and SOD levels. The remarkably higher positive rate of senescence-associated β-galactosidase and increased intracellular ROS levels in the HO treatment group were reversed by treatment with 10 µM RESV. As compared to the HO treatment group, 10 µM RESV could upregulate autophagy through the regulation of p-Rb, LC3, and p62 levels. The anti-aging effect of RESV via an autophagy regulation mechanism was further confirmed by the suppression of these effects with 3-MA treatment.

CONCLUSION

RESV may reverse and delay the aging process of HUVECs via upregulation of autophagy and could be a candidate therapeutic for age-related atherosclerotic diseases.

摘要

目的

白藜芦醇(RESV;反式-3,5,4'-三羟基芪)已成为一种治疗年龄相关性动脉粥样硬化疾病的潜在新疗法。然而,RESV对细胞衰老的影响及其潜在机制仍不清楚。因此,本研究旨在探讨RESV是否能通过上调自噬来延缓细胞衰老。

材料与方法

人脐静脉内皮细胞(HUVECs)分为四组:对照组、单独过氧化氢(HO)组、HO + RESV预处理组和HO + 3-甲基腺嘌呤(3-MA)+ RESV预处理干预组。采用细胞计数试剂盒-8法评估细胞活力。使用商业试剂盒检测超氧化物歧化酶(SOD)活性和细胞内活性氧(ROS)水平。通过免疫组织化学染色检测衰老相关的β-半乳糖苷酶活性。通过蛋白质印迹法检测有无RESV时衰老相关和自噬相关标志物的表达水平,包括磷酸化Rb(p-Rb)、微管相关蛋白1轻链3(LC3)和p62。

结果

10 μM RESV预处理可提高细胞活力和SOD水平。HO处理组中显著更高的衰老相关β-半乳糖苷酶阳性率和细胞内ROS水平升高被10 μM RESV处理所逆转。与HO处理组相比,10 μM RESV可通过调节p-Rb、LC3和p62水平上调自噬。3-MA处理抑制这些作用进一步证实了RESV通过自噬调节机制的抗衰老作用。

结论

RESV可能通过上调自噬来逆转和延缓HUVECs的衰老过程,可能成为治疗年龄相关性动脉粥样硬化疾病的候选疗法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43d9/6391141/e7693cace43b/dddt-13-747Fig1.jpg

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