黄芪多糖通过调节 SIRT-1/p53 信号通路减轻大鼠主动脉内皮细胞衰老。

Astragalus polysaccharides attenuate rat aortic endothelial senescence via regulation of the SIRT-1/p53 signaling pathway.

机构信息

Department of Endocrinology, The Second Medical Center & National Clinical Research Center for Geriatric Diseases, Chinese PLA General Hospital, Beijing, P.R. China.

Department of Nephrology, The First Medical Center, State Key Laboratory of Kidney Diseases, Chinese PLA General Hospital & Chinese PLA Institute of Nephrology, National Clinical Research Center for Kidney Diseases, Beijing, P.R. China.

出版信息

BMC Complement Med Ther. 2024 Feb 8;24(1):80. doi: 10.1186/s12906-024-04387-4.

DOI:10.1186/s12906-024-04387-4

PMID:38331805

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10851563/

Abstract

BACKGROUND

Astragalus polysaccharides (APS) have been verified to have antioxidative and antiaging activities in the mouse liver and brain. However, the effect of APS on aortic endothelial senescence in old rats and its underlying mechanism are currently unclear. Here, we aimed to elucidate the effects of APS on rat aortic endothelial oxidative stress and senescence in vitro and in vivo and investigate the potential molecular targets.

METHODS

Twenty-month-old natural aging male rats were treated with APS (200 mg/kg, 400 mg/kg, 800 mg/kg daily) for 3 months. Serum parameters were tested using corresponding assay kits. Aortic morphology was observed by staining with hematoxylin and eosin (H&E) and Verhoeff Van Gieson (VVG). Aging-related protein levels were evaluated using immunofluorescence and western blot analysis. Primary rat aortic endothelial cells (RAECs) were isolated by tissue explant method. RAEC mitochondrial function was evaluated by the mitochondrial membrane potential (MMP) measured with the fluorescent lipophilic cationic dye JC‑1. Intracellular total antioxidant capacity (T-AOC) was detected by a commercial kit. Cellular senescence was assessed using senescence-associated-β-galactosidase (SA-β-Gal) staining.

RESULTS

Treatment of APS for three months was found to lessen aortic wall thickness, renovate vascular elastic tissue, improve vascular endothelial function, and reduce oxidative stress levels in 20-month-old rats. Primary mechanism analysis showed that APS treatment enhanced Sirtuin 1 (SIRT-1) protein expression and decreased the levels of the aging marker proteins p53, p21 and p16 in rat aortic tissue. Furthermore, APS abated hydrogen peroxide (HO)-induced cell senescence and restored HO-induced impairment of the MMP and T-AOC in RAECs. Similarly, APS increased SIRT-1 and decreased p53, p21 and p16 protein levels in senescent RAECs isolated from old rats. Knockdown of SIRT-1 diminished the protective effect of APS against HO-induced RAEC senescence and T-AOC loss, increased the levels of the downstream proteins p53 and p21, and abolished the inhibitory effect of APS on the expression of these proteins in RAECs.

CONCLUSION

APS may reduce rat aortic endothelial oxidative stress and senescence via the SIRT-1/p53 signaling pathway.

摘要

背景

黄芪多糖（APS）已被证实具有抗氧化和抗衰老作用，可作用于小鼠的肝脏和大脑。然而，APS 对老年大鼠主动脉内皮细胞衰老的影响及其潜在机制尚不清楚。本研究旨在探讨 APS 对体外和体内大鼠主动脉内皮氧化应激和衰老的影响，并研究其潜在的分子靶点。

方法

20 月龄自然衰老雄性大鼠给予 APS（200mg/kg、400mg/kg、800mg/kg，每日 1 次）治疗 3 个月。采用相应的检测试剂盒检测血清参数。采用苏木精-伊红（H&E）和Verhoeff van Gieson（VVG）染色观察主动脉形态。采用免疫荧光和蛋白质印迹法检测衰老相关蛋白水平。采用组织块法分离原代大鼠主动脉内皮细胞（RAECs）。采用荧光亲脂性阳离子染料 JC-1 测量线粒体膜电位（MMP）来评估 RAEC 线粒体功能。采用商业试剂盒检测细胞内总抗氧化能力（T-AOC）。采用衰老相关-β-半乳糖苷酶（SA-β-Gal）染色评估细胞衰老。

结果

3 个月的 APS 治疗可减轻 20 月龄大鼠主动脉壁厚度，修复血管弹性组织，改善血管内皮功能，降低氧化应激水平。初步机制分析表明，APS 治疗可增强 Sirtuin 1（SIRT-1）蛋白表达，并降低大鼠主动脉组织中衰老标志物蛋白 p53、p21 和 p16 的水平。此外，APS 可减轻过氧化氢（HO）诱导的细胞衰老，并恢复 HO 诱导的 RAECs 中线粒体膜电位和 T-AOC 的损伤。同样，APS 可增加衰老的 RAEC 中 SIRT-1 蛋白的表达，降低 p53、p21 和 p16 蛋白的水平，而 SIRT-1 敲低则减弱了 APS 对 HO 诱导的 RAEC 衰老和 T-AOC 丢失的保护作用，增加了下游蛋白 p53 和 p21 的水平，并消除了 APS 对 RAECs 中这些蛋白表达的抑制作用。