LOX-1促进低氧诱导的内皮细胞自噬与凋亡

doi:10.12092/j.issn.1009-2501.2025.08.002

中国临床药理学与治疗学 ›› 2025, Vol. 30 ›› Issue (8): 1017-1025.doi: 10.12092/j.issn.1009-2501.2025.08.002

LOX-1促进低氧诱导的内皮细胞自噬与凋亡

李燕飞¹, 黄灿², 罗平³, 何芳⁴, 胡长平²

¹中南大学湘雅医学院附属长沙医院,长沙市第一医院,药剂科,长沙 410005,湖南;
²中南大学湘雅药学院药理学系,心血管研究湖南省重点实验室,长沙 410013,湖南;
³中南大学湘雅医院药学部,长沙 410008,湖南;
⁴南华大学附属南华医院教学科研部,衡阳 421002,湖南

收稿日期:2025-03-30 修回日期:2025-07-10 发布日期:2025-08-12
通讯作者: 胡长平,男,博士,教授,博士生导师,研究方向：心血管药理学。E-mail： huchangping@csu.edu.cn
作者简介:李燕飞,女,硕士,主管药师,研究方向：临床药学。E-mail： 919454256@qq.com
基金资助:
国家自然科学基金（82173817,82241025）; 湖南省卫生健康委科研计划课题资助项目（D202313016976）; 中国医药教育协会2023年“临床用药卫生技术评估”专项课题资助项目（2023WSJSPGZXKT-21）

LOX-1 promotes hypoxia-induced autophagy and apoptosis in endothelial cells

LI Yanfei, HUANG Can, LUO Ping, HE Fang, HU Changping

¹Department of Pharmacy, The Affiliated Changsha Hospital of Xiangya School of Medicine, Central South University; The First Hospital of Changsha, Changsha 410005, Hunan, China;
²Department of Pharmacology, Xiangya School of Pharmaceutical Sciences, Central South University; Hunan Provincial Key Laboratory of Cardiovascular Research, Changsha 410013, Hunan, China;
³Department of Pharmacy, Xiangya Hospital, Central South University, Changsha 410008, Hunan, China;
⁴Department of Teaching and Research, The Affiliated Nanhua Hospital of Nanhua University, Hengyang 421002, Hunan, China

Received:2025-03-30 Revised:2025-07-10 Published:2025-08-12

摘要/Abstract

摘要： 目的：探讨低氧环境下植物凝集素样氧化性低密度脂蛋白受体-1（lectin-like oxidized low-density lipoprotein receptor-1,LOX-1）对内皮细胞自噬和凋亡的影响。方法：将人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）分别给予低氧（1% O₂）不同时间（0、6、12、24 h）处理后,观察细胞的自噬和凋亡水平。进一步通过干预LOX-1以探究其对自噬和凋亡的影响。采用荧光显微镜观察内皮细胞感染GFP-LC3B腺病毒后LC3B的表达;透射电镜检测自噬小体;Real-time PCR法检测LOX-1 mRNA水平;Western blot检测LOX-1、LC3II/I、Beclin-1、Atg 5、cleaved-caspase 3、Bax、Bcl-2蛋白表达;DCFH-DA荧光探针检测活性氧（reactive oxygen species,ROS）水平;Hoechst法、流式细胞术（Annexin V-PI双染）检测细胞凋亡。结果：低氧（1% O₂）处理HUVECs 24 h,可明显增加荧光显微镜下LC3II斑点,上调LC3II/I蛋白表达,电镜下检测到自噬小体;低氧增加ROS生成,促进细胞凋亡;低氧上调HUVECs LOX-1 mRNA和蛋白的表达;LOX-1 siRNA可显著逆转低氧引起的细胞自噬,自噬相关蛋白（Beclin-1、Atg 5、LC3II/I）表达下调;LOX-1 siRNA能减少ROS生成,抑制细胞凋亡,促凋亡蛋白cleaved-caspase 3、Bax的表达下调,抑凋亡蛋白Bcl-2表达上调。结论：低氧诱导LOX-1表达上调,促进ROS生成,从而介导内皮细胞自噬与凋亡。

关键词: 植物凝集素样氧化性低密度脂蛋白受体-1, 低氧, 自噬, 凋亡, 内皮细胞

Abstract: AIM: To investigate the role of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in hypoxia-induced autophagy and apoptosis in endothelial cells. METHODS: Human umbilical vein endothelial cells (HUVECs) were exposed to hypoxia (1%O₂) for varying durations (0, 6, 12, 24 h) to evaluate autophagy and apoptosis levels. LOX-1 was further intervened to explore its effects on autophagy and apoptosis. GFP-LC3B adenovirus infection was observed under fluorescence microscopy to assess LC3B expression. Autophagosomes were detected by transmission electron microscopy (TEM). LOX-1 mRNA levels were measured using real-time PCR. Protein expression of LOX-1, LC3II/I, Beclin-1, Atg5, cleaved-caspase 3, Bax, and Bcl-2 was analyzed by Western blot. Reactive oxygen species (ROS) levels were quantified using the DCFH-DA fluorescent probe. Apoptosis was assessed via Hoechst staining and flow cytometry (Annexin V-PI double staining). RESULTS: Hypoxia (1%O₂, 24 h) significantly increased LC3II puncta under fluorescence microscopy, upregulated LC3II/I protein expression, and induced autophagosome formation observed by TEM. Hypoxia elevated ROS production and promoted apoptosis. LOX-1 mRNA and protein expression were upregulated in hypoxic HUVECs. LOX-1 siRNA intervention markedly reversed hypoxia-induced autophagy, downregulating autophagy-related proteins (Beclin-1, Atg5, LC3II/I). LOX-1 siRNA also suppressed ROS generation and inhibited apoptosis, as evidenced by decreased expression of cleaved-caspase 3 and Bax, and increased Bcl-2 levels. CONCLUSION: Hypoxia induced upregulation of LOX-1 expression to produce ROS, thereby promoting autophagy and apoptosis in endothelial cells.

Key words: lectin-like oxidized low-density lipoprotein receptor-1, hypoxia, autophagy, apoptosis, endothelial cell

中图分类号:

R965.2

李燕飞, 黄灿, 罗平, 何芳, 胡长平. LOX-1促进低氧诱导的内皮细胞自噬与凋亡[J]. 中国临床药理学与治疗学, 2025, 30(8): 1017-1025.

LI Yanfei¹, HUANG Can², LUO Ping³, HE Fang⁴, HU Changping². LOX-1 promotes hypoxia-induced autophagy and apoptosis in endothelial cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2025, 30(8): 1017-1025.

参考文献

[1] Wu JB, Lei Z, Yu JG.Hypoxia induces autophagy in human vascular endothelial cells in a hypoxia-inducible factor 1-dependent manner[J]. Mol Med Rep, 2015, 11(4): 2677-2682.
[2] Carmeliet P.Mechanisms of angiogenesis and arteriogenesis[J]. Nat Med, 2000, 6(4): 389-395.
[3] Janaszak-Jasiecka A, Siekierzycka A, Płoska A, et al.Endothelial dysfunction driven by hypoxia-the influence of oxygen deficiency on NO bioavailability[J]. Biomolecules, 2021, 11(7): 982.
[4] Baldea I, Teacoe I, Olteanu DE, et al.Effects of different hypoxia degrees on endothelial cell cultures-Time course study[J]. Mech Ageing Dev, 2018, 172: 45-50.
[5] Fernández A, Ordóñez R, Reiter RJ, et al.Melatonin and endoplasmic reticulum stress: relation to autophagy and apoptosis[J]. J Pineal Res, 2015, 59(3): 292-307.
[6] Ren H, Dai RC, Nik Nabil WN, et al.Unveiling the dual role of autophagy in vascular remodelling and its related diseases[J]. Biomed Pharmacother, 2023, 168: 115643.
[7] Lin XY, Ouyang SY, Zhi CX, et al.Focus on ferroptosis, pyroptosis, apoptosis and autophagy of vascular endothelial cells to the strategic targets for the treatment of atherosclerosis[J]. Arch Biochem Biophys, 2022, 715: 109098.
[8] Zhao FJ, Satyanarayana G, Zhang Z, et al.Endothelial autophagy in coronary microvascular dysfunction and cardiovascular disease[J]. Cells, 2022, 11(13): 2081.
[9] Taye A, El-Sheikh AA.Lectin-like oxidized low-density lipoprotein receptor 1 pathways[J]. Eur J Clin Invest, 2013, 43(7): 740-745.
[10] Yan ML, Mehta JL, Zhang WF, et al.LOX-1, oxidative stress and inflammation: a novel mechanism for diabetic cardiovascular complications[J]. Cardiovasc Drugs Ther, 2011, 25(5): 451-459.
[11] Ding ZF, Liu SJ, Deng XY, et al.Hemodynamic shear stress modulates endothelial cell autophagy: Role of LOX-1[J]. Int J Cardiol, 2015, 184: 86-95.
[12] Duan QD, Si HL, Tian LM, et al.LOX-1 attenuates high glucose-induced autophagy via AMPK/HNF4α signaling in HLSECs[J]. Heliyon, 2022, 8(12): e12385.
[13] Mollace V, Gliozzi M, Musolino V, et al.Oxidized LDL attenuates protective autophagy and induces apoptotic cell death of endothelial cells: Role of oxidative stress and LOX-1 receptor expression[J]. Int J Cardiol, 2015, 184: 152-158.
[14] Luo P, Zhang WF, Qian ZX, et al.MiR-590-5p-meidated LOX-1 upregulation promotes angiotensin II-induced endothelial cell apoptosis[J]. Biochem Biophys Res Commun, 2016, 471(4): 402-408.
[15] Liu MX, Galli G, Wang YL, et al.Novel therapeutic targets for hypoxia-related cardiovascular diseases: the role of HIF-1[J]. Front Physiol, 2020, 11: 774.
[16] Pan CY, Ai CY, Liang LL, et al.Sestrin2 protects against hypoxic nerve injury by regulating mitophagy through SESN2/AMPK pathway[J]. Front Mol Biosci, 2023, 10: 1266243.
[17] Liu YE, Sun YH, Guo YD, et al.An overview: the diversified role of mitochondria in cancer metabolism[J]. Int J Biol Sci, 2023, 19(3): 897-915.
[18] Chang NC.Autophagy and stem cells: Self-eating for self-renewal[J]. Front Cell Dev Biol, 2020, 8: 138.
[19] Li MM, Tan J, Miao YY, et al.The dual role of autophagy under hypoxia-involvement of interaction between autophagy and apoptosis[J]. Apoptosis, 2015, 20(6): 769-777.
[20] He XM, Wu F, Zhou Y, et al.miR-4463 regulates hypoxia-induced autophagy and apoptosis by targeting ULK1 in endothelial cells[J]. Front Biosci (Landmark Ed), 2022, 27(6): 175.
[21] Tao ZQ, Wei BZ, Zhao M, et al.Hypoxia affects autophagy in human umbilical vein endothelial cells via the IRE1 unfolded protein response[J]. Curr Med Sci, 2023, 43(4): 689-695.
[22] Xu SW, Ogura S, Chen JW, et al.LOX-1 in atherosclerosis: biological functions and pharmacological modifiers[J]. Cell Mol Life Sci, 2013, 70(16): 2859-2872.
[23] Zhou XL, Chen XF, Zhang L, et al.Mannose-binding lectin reduces oxidized low-density lipoprotein induced vascular endothelial cells injury by inhibiting LOX1-ox-LDL binding and modulating autophagy[J]. Biomedicines, 2023, 11(6): 1743.
[24] 朱惠莲, 唐志红, 刘静, 等. Ox-LDL诱导血管内皮细胞表达LOX-1[J]. 中山大学学报(医学科学版), 2005, 26(6): 612-616.
[25] Ding ZF, Liu SJ, Sun CQ, et al.Concentration polarization of ox-LDL activates autophagy and apoptosis via regulating LOX-1 expression[J]. Sci Rep, 2013, 3: 2091.
[26] Barbieri E, Sestili P.Reactive oxygen species in skeletal muscle signaling[J]. J Signal Transduct, 2012, 2012: 982794.
[27] Odinokova IV, Sung KF, Mareninova OA, et al.Mechanisms regulating cytochrome C release in pancreatic mitochondria[J]. GUT, 2009, 58(3): 431-442.
[28] Scherz-Shouval R, Shvets E, Fass E, et al.Reactive oxygen species are essential for autophagy and specifically regulate the activity of Atg4[J]. EMBO J, 2019, 38(10): e101812.
[29] Lubrano V, Balzan S.LOX-1 and ROS, inseparable factors in the process of endothelial damage[J]. Free Radic Res, 2014, 48(8): 841-848.
[30] Chen ZB, Wang M, He Q, et al.MicroRNA-98 rescues proliferation and alleviates ox-LDL-induced apoptosis in HUVECs by targeting LOX-1[J]. Exp Ther Med, 2017, 13(5): 1702-1710.

LOX-1促进低氧诱导的内皮细胞自噬与凋亡

LOX-1 promotes hypoxia-induced autophagy and apoptosis in endothelial cells

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