基于细胞提取/ UPLC-MS /MS的桑枝ox-LDL抗动脉粥样硬化化合物鉴定及其机制分析

IF 5.2 2区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Zhihui Jin, Wei Xiang, Xiaowen Shi, Fenglin Jiang, Yanan Jia, Yuansong Zhang, Lingshu Zeng, Xianzhi Huang, Li Xu
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引用次数: 0

摘要

桑枝是一种传统的中药和农业副产品,含有具有抗动脉粥样硬化潜力的生物活性化合物。本研究旨在鉴定和评估桑枝提取物(MTEs)中关键化合物对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)的影响,重点了解这些化合物如何调节氧化应激及其相关信号通路。方法采用生物特异性细胞提取和超高效液相色谱-串联质谱(UHPLC-MS /MS)技术对这些化合物进行筛选鉴定。通过测量细胞活力、丙二醛(MDA)和超氧化物歧化酶(SOD)水平,以及使用2,7 -二氯二氢荧光素二乙酸酯(DCFH-DA)和二氢乙胺(DHE)探针检测细胞内活性氧(ROS)来评估保护作用。采用Real-time qPCR和Western blotting分析mRNA和蛋白水平。结果鉴定出两种新的活性化合物:苦参H和Morusin,以及已知的苦参G。在ox- ldl处理的huvec中,它们显著降低MDA和ROS水平,同时增加SOD活性。Kuwanon H尤其有效,可增强核因子红系2相关因子2 (Nrf-2)活性,上调其靶基因血红素加氧酶-1 (HO-1)和NAD(P)H:醌氧化还原酶1 (nqc -1)。结论Kuwanon H、Morusin和Kuwanon G对ox- ldl诱导的HUVECs氧化损伤具有保护作用,其中Kuwanon H通过Nrf-2/HO-1信号通路表现出最强的保护作用。这些化合物在治疗动脉粥样硬化和相关氧化应激疾病方面具有潜力。图形抽象
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis

Background

Mulberry twigs, a traditional Chinese medicinal and agricultural byproduct, contain bioactive compounds with anti-atherosclerotic potential. This study aims to identify and evaluate the effects of key compounds in mulberry twig extracts (MTEs) on oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cells (HUVECs), with a focus on understanding how these compounds modulate oxidative stress and related signaling pathways.

Methods

Biospecific cell extraction and ultra-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) were employed to screen and identify these compounds. Protective effects were assessed by measuring cell viability, malondialdehyde (MDA), and superoxide dismutase (SOD) levels, along with detecting intracellular reactive oxygen species (ROS) using 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) and dihydroethidine (DHE) probes. Real-time qPCR and Western blotting were used for mRNA and protein level analysis.

Results

Two novel active compounds, Kuwanon H and Morusin, and the known Kuwanon G, were identified. They significantly reduced MDA and ROS levels while increasing SOD activity in ox-LDL-treated HUVECs. Kuwanon H was particularly effective, enhancing nuclear factor erythroid 2-related factor 2 (Nrf-2) activity and upregulating its target genes Heme oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO-1).

Conclusion

In conclusion, Kuwanon H, Morusin, and Kuwanon G effectively protected HUVECs from ox-LDL-induced oxidative injury, with Kuwanon H showing the strongest protective effects via the Nrf-2/HO-1 signaling pathway. These compounds hold potential in treating atherosclerosis and related oxidative stress diseases.

Graphical Abstract

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来源期刊
Chemical and Biological Technologies in Agriculture
Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.80
自引率
3.00%
发文量
83
审稿时长
15 weeks
期刊介绍: Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.
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