Abstract
This study aims to investigate the impact of hepatocyte nuclear factor 1β (HNF1b) on macrophage sortilin-mediated lipid metabolism and aortic atherosclerosis and explore the role of the flavone of Polygonatum odoratum (PAOA-flavone)-promoted small ubiquitin-related modifier (SUMO) modification in the atheroprotective efficacy of HNF1b. HNF1b was predicted to be a transcriptional regulator of sortilin expression via bioinformatics, dual-luciferase reporter gene assay, and chromatin immunoprecipitation. HNF1b overexpression decreased sortilin expression and cellular lipid contents in THP-1 macrophages, leading to a depression in atherosclerotic plaque formation in low-density lipoprotein (LDL) receptor-deficient (LDLR−/−) mice. Multiple SUMO1-modified sites were identified on the HNF1b protein and co-immunoprecipitation confirmed its SUMO1 modification. The SUMOylation of HNF1b protein enhanced the HNF1b-inhibited effect on sortilin expression and reduced lipid contents in macrophages. PAOA-flavone treatment promoted SUMO-activating enzyme subunit 1 (SAE1) expression and SAE1-catalyzed SUMOylation of the HNF1b protein, which prevented sortilin-mediated lipid accumulation in macrophages and the formation of atherosclerotic plaques in apolipoprotein E-deficient (ApoE−/−) mice. Interference with SAE1 abrogated the improvement in lipid metabolism in macrophage cells and atheroprotective efficacy in vivo upon PAOA-flavone administration. In summary, HNF1b transcriptionally suppressed sortilin expression and macrophage lipid accumulation to inhibit aortic lipid deposition and the development of atherosclerosis. This anti-atherosclerotic effect was enhanced by PAOA-flavone-facilitated, SAE1-catalyzed SUMOylation of the HNF1b protein.
摘要
本研究旨在探究肝细胞核因子1β(HNF1b)对巨噬细胞分拣蛋白(sortilin)介导的脂质代谢和主动脉粥样硬化的影响,以及玉竹(Polygonatumodoratum)黄酮在促进HNF1b的小泛素蛋白(SUMO)化修饰在动脉粥样硬化保护功效中的作用。通过生物信息学、双荧光素酶报告基因分析和染色质免疫共沉淀试验预测了HNF1b是sortilin表达的转录调控因子。HNF1b过表达降低了人髓系白血病单核(THP-1)巨噬细胞中sortilin的表达和细胞内脂质含量,从而抑制了低密度脂蛋白受体基因敲除(LDLR−/−)小鼠的动脉粥样斑块形成。在HNF1b蛋白上鉴定出多个SUMO1修饰位点,并通过免疫共沉淀证实存在SUMO1修饰。HNF1b蛋白的SUMO化修饰增强了HNF1b对sortilin表达的抑制作用,并降低了巨噬细胞中的脂质含量。玉竹黄酮处理促进了SUMO活化酶E1(SAE1)的表达和SAE1催化下对HNF1b蛋白的SUMO化修饰,从而阻止了巨噬细胞中sortilin介导的脂质积累和载脂蛋白E基因敲除(ApoE−/−)小鼠的动脉粥样斑块形成,而对SAE1的干扰使得巨噬细胞内脂质代谢的改善和玉竹黄酮治疗对体内抗动脉粥样硬化效果消失。因此,HNF1b通过转录抑制sortilin表达和巨噬细胞内脂质积累,抑制主动脉脂质沉积和动脉粥样硬化的发展,玉竹黄酮促进SAE1催化的HNF1b蛋白SUMO化修饰增强了其抗动脉粥样硬化效应。
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Data availability statement
The original contributions presented in the study are included in the article/supplementary materials.
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Acknowledgments
This study was supported by the Natural Science Foundation of Guangxi Zhuang Autonomous Region, China (No. 2019JJA140728), the Horizontal Cooperation Project with Hunan Mingshun Pharmaceutical Co., Ltd. (No. 2021GLHX02), and the Guangxi Province Postgraduate Co-training Base for Cooperative Innovation in Basic Medicine of Guilin Medical University (No. Gui Xue Wei [2020]7). The authors thank Sha SUN (Union Shenzhen Hospital, Shenzhen, China) for the support in pilot research, Feng YAO (Guangxi Key Laboratory of Diabetic Systems Medicine, Faculty of Basic Medical Sciences, Guilin Medical University, Guilin, China) for the technical guidance in animal husbandry and treatment, and Zhifeng LONG (Microscopic Morphology Experimental Center, School of Basic Medicine, University of South China, Hengyang, China) for the assistance in histological slicing and staining.
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Fang LIU performed the experimental research and data analysis, wrote and edited the manuscript. Shirui CHEN and Xinyue MING performed the establishment of animal models. Huijuan LI performed the experiments and analyzed the data. Zhaoming ZENG providied us the samples of PAOA-flavone and gave some advice for its use. Yuncheng LV contributed to the research design and writing and editing of the manuscript. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Fang LIU, Shirui CHEN, Xinyue MING, Huijuan LI, Zhaoming ZENG, and Yuncheng LV declare that they have no conflict of interest.
All the animal experiments were performed by following the Guidelines for Laboratory Animals of the National Institute of Health and approved by the Committee on the Ethics of Animal Experiments of Guilin Medical University (No. GLMC-IACUC-2022011).
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Figs. S1 and S2; Table S1; Materials and methods
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Liu, F., Chen, S., Ming, X. et al. Sortilin-induced lipid accumulation and atherogenesis are suppressed by HNF1b SUMOylation promoted by flavone of Polygonatum odoratum. J. Zhejiang Univ. Sci. B 24, 998–1013 (2023). https://doi.org/10.1631/jzus.B2200682
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DOI: https://doi.org/10.1631/jzus.B2200682
Key words
- Atherosclerosis
- Lipid accumulation
- Hepatocyte nuclear factor 1β (HNF1b)
- Flavone of Polygonatum odoratum
- SUMOylation