Yiqihuoxue decoction (GSC) inhibits mitochondrial fission through the AMPK pathway to ameliorate EPCs senescence and optimize vascular aging transplantation regimens.

IF 5.3 3区 医学 Q1 INTEGRATIVE & COMPLEMENTARY MEDICINE
Yinan Liu, Zenghui Niu, Xue Wang, Chengkui Xiu, Yanhong Hu, Jiali Wang, Yan Lei, Jing Yang
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Abstract

Background: During the aging process, the number and functional activity of endothelial progenitor cells (EPCs) are impaired, leading to the unsatisfactory efficacy of transplantation. Previous studies demonstrated that Yiqihuoxue decoction (Ginseng-Sanqi-Chuanxiong, GSC) exerts anti-vascular aging effects. The purpose of this study is to evaluated the effects of GSC on D-galactose (D-gal)induced senescence and the underlying mechanisms.

Methods: The levels of cellular senescence-related markers P16, P21, P53, AMPK and p-AMPK were detected by Western blot analysis (WB). SA-β-gal staining was used to evaluate cell senescence. EPCs function was measured by CCK-8, Transwell cell migration and cell adhesion assay. The morphological changes of mitochondria were detected by confocal microscopy. The protein and mRNA expression of mitochondrial fusion fission Drp1, Mff, Fis1, Mfn1, Mfn2 and Opa1 in mitochondria were detect using WB and RT-qPCR. Mitochondrial membrane potential, mtROS and ATP of EPCs were measured using IF. H&E staining was used to observe the pathological changes and IMT of the aorta. The expressions of AGEs, MMP-2 and VEGF in aorta were measured using Immunohistochemical (IHC). The levels of SOD, MDA, NO and ET-1 in serum were detected by SOD, MDA and NO kits.

Results: In vitro, GSC ameliorated the senescence of EPCs induced by D-gal and reduced the expression of P16, P21 and P53. The mitochondrial morphology of EPCs was restored, the expression of mitochondrial Drp1, Mff and Fis1 protein was decreased, the levels of mtROS and ATP were decreased, and mitochondrial function was improved. Meanwhile, the expression of AMPK and p-AMPK increased. The improvement effects of GSC on aging and mitochondrial morphology and function were were hindered after adding AMPK inhibitor. In vivo, GSC improved EPCs efficiency, ameliorated aortic structural disorder and decreased IMT in aging mice. The serum SOD level increased and MDA level decreased, indicating the improvement of antioxidant capacity. Increased NO content and ET-1 content suggested improvement of vascular endothelial function. The changes observed in SOD and MMP-2 suggested a reduction in vascular stiffness and the degree of vascular damage. The decreased expression of P21 and P53 indicates the delay of vascular senescence.

益气活血汤(GSC)通过AMPK途径抑制线粒体分裂,从而改善EPCs衰老,优化血管衰老移植方案。
背景:在衰老过程中,内皮祖细胞(EPCs)的数量和功能活性会受损,导致移植效果不理想。以往的研究表明,益气补血汤(人参-三七-川芎汤)具有抗血管衰老的作用。本研究旨在评估人参三七川芎汤对D-半乳糖(D-gal)诱导的衰老的影响及其内在机制:方法:细胞衰老相关标志物 P16、P21、P53、AMPK 和 p-AMPK 的水平通过 Western 印迹分析(WB)进行检测。SA-β-gal染色用于评估细胞衰老。EPCs功能通过CCK-8、Transwell细胞迁移和细胞粘附试验进行检测。共聚焦显微镜检测线粒体的形态变化。利用 WB 和 RT-qPCR 检测线粒体中线粒体融合裂变 Drp1、Mff、Fis1、Mfn1、Mfn2 和 Opa1 的蛋白和 mRNA 表达。使用 IF 检测 EPCs 的线粒体膜电位、mtROS 和 ATP。用 H&E 染色观察主动脉的病理变化和内径。用免疫组化(IHC)方法检测主动脉中 AGEs、MMP-2 和 VEGF 的表达。用 SOD、MDA 和 NO 试剂盒检测血清中 SOD、MDA、NO 和 ET-1 的水平:结果:在体外,GSC能改善D-gal诱导的EPCs衰老,并减少P16、P21和P53的表达。EPCs 的线粒体形态得到恢复,线粒体 Drp1、Mff 和 Fis1 蛋白表达减少,mtROS 和 ATP 水平下降,线粒体功能得到改善。同时,AMPK 和 p-AMPK 的表达增加。加入 AMPK 抑制剂后,GSC 对衰老和线粒体形态及功能的改善作用受到阻碍。在体内,GSC 提高了 EPCs 的效率,改善了衰老小鼠的主动脉结构紊乱并降低了 IMT。血清 SOD 水平升高,MDA 水平降低,表明抗氧化能力提高。NO含量和ET-1含量的增加表明血管内皮功能得到改善。观察到的 SOD 和 MMP-2 的变化表明血管僵硬度和血管损伤程度有所降低。P21 和 P53 表达的减少表明血管衰老的延迟。
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来源期刊
Chinese Medicine
Chinese Medicine INTEGRATIVE & COMPLEMENTARY MEDICINE-PHARMACOLOGY & PHARMACY
CiteScore
7.90
自引率
4.10%
发文量
133
审稿时长
31 weeks
期刊介绍: Chinese Medicine is an open access, online journal publishing evidence-based, scientifically justified, and ethical research into all aspects of Chinese medicine. Areas of interest include recent advances in herbal medicine, clinical nutrition, clinical diagnosis, acupuncture, pharmaceutics, biomedical sciences, epidemiology, education, informatics, sociology, and psychology that are relevant and significant to Chinese medicine. Examples of research approaches include biomedical experimentation, high-throughput technology, clinical trials, systematic reviews, meta-analysis, sampled surveys, simulation, data curation, statistics, omics, translational medicine, and integrative methodologies. Chinese Medicine is a credible channel to communicate unbiased scientific data, information, and knowledge in Chinese medicine among researchers, clinicians, academics, and students in Chinese medicine and other scientific disciplines of medicine.
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