Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis
Abstract 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...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2025-01-01
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| Series: | Chemical and Biological Technologies in Agriculture |
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| Online Access: | https://doi.org/10.1186/s40538-024-00709-3 |
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| author | Zhihui Jin Wei Xiang Xiaowen Shi Fenglin Jiang Yanan Jia Yuansong Zhang Lingshu Zeng Xianzhi Huang Li Xu |
| author_facet | Zhihui Jin Wei Xiang Xiaowen Shi Fenglin Jiang Yanan Jia Yuansong Zhang Lingshu Zeng Xianzhi Huang Li Xu |
| author_sort | Zhihui Jin |
| collection | DOAJ |
| description | Abstract 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 |
| format | Article |
| id | doaj-art-78ffa32b93c249e08a774ac27d6c2642 |
| institution | Kabale University |
| issn | 2196-5641 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Chemical and Biological Technologies in Agriculture |
| spelling | doaj-art-78ffa32b93c249e08a774ac27d6c26422025-01-19T12:13:02ZengSpringerOpenChemical and Biological Technologies in Agriculture2196-56412025-01-0112111210.1186/s40538-024-00709-3Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysisZhihui Jin0Wei Xiang1Xiaowen Shi2Fenglin Jiang3Yanan Jia4Yuansong Zhang5Lingshu Zeng6Xianzhi Huang7Li Xu8State Key Laboratory of Resource Insects, College of Sericulture, Textile and Biomass Sciences, Southwest UniversityChongqing University of Chinese MedicineThe Second Affiliated Hospital of Jiaxing UniversityState Key Laboratory of Resource Insects, College of Sericulture, Textile and Biomass Sciences, Southwest UniversityState Key Laboratory of Resource Insects, College of Sericulture, Textile and Biomass Sciences, Southwest UniversityState Key Laboratory of Resource Insects, College of Sericulture, Textile and Biomass Sciences, Southwest UniversityChongqing Sericulture Science and Technology Research InstituteState Key Laboratory of Resource Insects, College of Sericulture, Textile and Biomass Sciences, Southwest UniversityState Key Laboratory of Resource Insects, College of Sericulture, Textile and Biomass Sciences, Southwest UniversityAbstract 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 Abstracthttps://doi.org/10.1186/s40538-024-00709-3Mulberry twigsUPLC–MS/MSKuwanon HOxidative stressAnti-atherosclerosisNrf-2 |
| spellingShingle | Zhihui Jin Wei Xiang Xiaowen Shi Fenglin Jiang Yanan Jia Yuansong Zhang Lingshu Zeng Xianzhi Huang Li Xu Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis Chemical and Biological Technologies in Agriculture Mulberry twigs UPLC–MS/MS Kuwanon H Oxidative stress Anti-atherosclerosis Nrf-2 |
| title | Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis |
| title_full | Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis |
| title_fullStr | Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis |
| title_full_unstemmed | Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis |
| title_short | Identification of compounds against atherosclerosis induced by ox-LDL based on cell extraction/UPLC–MS/MS from mulberry twigs and their mechanistic analysis |
| title_sort | identification of compounds against atherosclerosis induced by ox ldl based on cell extraction uplc ms ms from mulberry twigs and their mechanistic analysis |
| topic | Mulberry twigs UPLC–MS/MS Kuwanon H Oxidative stress Anti-atherosclerosis Nrf-2 |
| url | https://doi.org/10.1186/s40538-024-00709-3 |
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