层流切应力通过上调自噬水平抑制氧化型低密度脂蛋白诱导的内皮祖细胞损伤作用

陈丽芳; 黄于朗; 陈俊羽; 罗新林

PDF(3204 KB)

湖南师范大学学报医学版 ›› 2024, Vol. 21 ›› Issue (1) : 26-33.

基础医学

层流切应力通过上调自噬水平抑制氧化型低密度脂蛋白诱导的内皮祖细胞损伤作用

陈丽芳¹, 黄于朗¹, 陈俊羽², 罗新林²

作者信息 +

Protective effect of shearstress on LDL-induced endothelial progenitor cell injury through up-regulation of autophagy

CHEN Lifang¹, HUANG Yulang¹, CHEN Junyu², LUO Xinlin²

Author information +

文章历史 +

摘要

目的分析层流切应力（LSS）对氧化型低密度脂蛋白（ox-LDL）诱导内皮祖细胞（EPCs）损伤的保护机制。方法通过CCK8分析EPCs中ox-LDL和LSS的处理条件;Transwell和成管实验分析LSS对ox-LDL处理前后EPCs细胞迁移和成管能力的影响;Western检测LSS和ox-LDL处理对细胞自噬、内皮-间质转化标志物的影响;Western分析自噬激活和抑制剂对ox-LDL和LSS处理后自噬、内皮-间质转化标志物的影响。结果 ox-LDL处理后EPCs细胞的增殖能力,迁移能力和成管能力均下调,LSS处理后显著升高,LSS处理对ox-LDL诱导的EPCs细胞损伤有保护作用。分子机制方面,LSS处理后EPCs细胞自噬水平升高。ox-LDL处理后EPCs细胞发生显著的内皮间质化转变,LSS处理可明显抑制这一过程。自噬激活剂和抑制剂验证了这些结果。结论 LSS可通过上调自噬水平抑制ox-LDL诱导的EPCs内皮间质化,对EPCs细胞起保护作用。

Abstract

Objective To analyze the protective mechanism of laminar shear stress (LSS) on endothelial progenitor cells (EPCs) injury induced by oxidized low-density lipoprotein (ox-LDL). Methods The treatment conditions of ox-LDL and LSS in EPCs were analyzed by CCK8. The effects of LSS on the migration and tube formation ability of EPCs before and after treatment with ox-LDL were analyzed by Transwell and tube formation experiments. The effects of LSS and ox-LDL treatment on autophagy and endothelial-mesenchymal transition markers were analyzed by Western blot. The effects of autophagy activators and inhibitors on autophagy and endothelial-mesenchymal transition markers after treatment with ox-LDL and LSS were analyzed by Western blot. Results After treatment with ox-LDL, the proliferation, migration, and tube formation ability of EPCs were down-regulated, and LSS treatment significantly increased them. LSS treatment had a protective effect on ox-LDL-induced EPCs cell injury. In terms of molecular mechanisms, LSS treatment increased the autophagy level of EPCs cells. After treatment with ox-LDL, EPCs cells underwent significant endothelial mesenchymal transition, which was significantly inhibited by LSS treatment. Autophagy activators and inhibitors confirmed these results. Conclusion LSS can inhibit ox-LDL-induced endothelial mesenchymal transformation of EPCs by upregulating autophagy, and plays a protective role in EPCs.

导出引用

陈丽芳, 黄于朗, 陈俊羽, 罗新林. 层流切应力通过上调自噬水平抑制氧化型低密度脂蛋白诱导的内皮祖细胞损伤作用[J]. 湖南师范大学学报医学版. 2024, 21(1): 26-33

CHEN Lifang, HUANG Yulang, CHEN Junyu, LUO Xinlin. Protective effect of shearstress on LDL-induced endothelial progenitor cell injury through up-regulation of autophagy[J]. Journal of Hunan Normal University(Medical Science). 2024, 21(1): 26-33

中图分类号： R363

参考文献

[1] 中国心血管健康与疾病报告编写组. 中国心血管健康与疾病报告2022 概要[J]. 中国循环杂志, 2023, 38(06): 583-612.
[2] THIJSSEN D H J, BRUNO R M, VAN MIL A, et al. Expert consensus and evidence-based recommendations for the assessment of flow-mediated dilation in humans[J]. Eur Heart J, 2019, 40(30): 2534-2547.
[3] TAMARGO I A, BAEK K I, KIM Y, et al.Flow-induced reprogramming of endothelial cells in atherosclerosis[J]. Nat Rev Cardiol, 2023, 20(11): 738-753.
[4] PSALTIS P J, SIMARI R D.Vascular wall progenitor cells in health and disease[J]. Circ Res, 2015, 116(8): 1392-1412.
[5] KIM H M, KIM K J, MOON J H, et al.Association between EPCs count and rate of coronary revascularization in asymptomatic type 2 diabetic patients[J]. Acta Diabetol, 2012, 49(6): 413-420.
[6] BETTENCOURT N, OLIVEIRA S, TOSCHKE A M, et al.Predictors of circulating endothelial progenitor cell levels in patients without known coronary artery disease referred for multidetector computed tomography coronary angiography[J]. Rev Port Cardiol, 2011, 30(10): 753-760.
[7] WANG X, SHEN Y, SHANG M, et al.Endothelial mechanobiology in atherosclerosis[J]. Cardiovasc Res, 2023, 119(8): 1656-1675.
[8] ZHOU J, LI Y S, CHIEN S.Shear stress-initiated signaling and its regulation of endothelial function[J]. Arterioscler Thromb Vasc Biol, 2014, 34(10): 2191-2198.
[9] CHENG H, ZHONG W, WANG L, et al.Effects of shear stress on vascular endothelial functions in atherosclerosis and potential therapeutic approaches[J]. Biomed Pharmacother, 2023, 158: 114198.
[10] DE MEYER G R, GROOTAERT M O, MICHIELS C F, et al. Autophagy in vascular disease[J]. Circ Res, 2015, 116(3): 468-479.
[11] ZHI K, LI M, BAI J, et al.Quercitrin treatment protects endothelial progenitor cells from oxidative damage via inducing autophagy through extracellular signal-regulated kinase[J]. Angiogenesis, 2016, 19(3): 311-324.
[12] WU S H, ZHANG F, YAO S, et al.Shear Stress Triggers Angiogenesis of Late Endothelial Progenitor Cells via the PTEN/Akt/GTPCH/BH4 Pathway[J]. Stem Cells Int, 2020, 2020: 5939530.
[13] STRANDBERG T E, KOLEHMAINEN L, VUORIO A.Evaluation and treatment of older patients with hypercholesterolemia: a clinical review[J]. JAMA, 2014, 312(11): 1136-1144.
[14] JIANG F, YANG J, ZHANG Y, et al.Angiotensin-converting enzyme 2 and angiotensin 1-7: novel therapeutic targets[J]. Nat Rev Cardiol, 2014, 11(7): 413-426.
[15] KARMALI K N, LLOYD-JONES D M, BERENDSEN M A, et al. Drugs for Primary Prevention of Atherosclerotic Cardiovascular Disease: An Overview of Systematic Reviews[J]. JAMA Cardiol, 2016, 1(3): 341-349.
[16] TOUSOULIS D, SIMOPOULOU C, PAPAGEORGIOU N, et al.Endothelial dysfunction in conduit arteries and in microcirculation. Novel therapeutic approaches[J]. Pharmacol Ther, 2014, 144(3): 253-267.
[17] XU S, ILYAS I, LITTLE P J, et al.Endothelial Dysfunction in Atherosclerotic Cardiovascular Diseases and Beyond: From Mechanism to Pharmacotherapies[J]. Pharmacol Rev, 2021, 73(3): 924-967.
[18] OESTERLE A, LAUFS U, LIAO J K.Pleiotropic Effects of Statins on the Cardiovascular System[J]. Circ Res, 2017, 120(1): 229-243.
[19] DOUGLAS G, VAN KAMPEN E, HALE A B, et al.Endothelial cell repopulation after stenting determines in-stent neointima formation: effects of bare-metal vs. drug-eluting stents and genetic endothelial cell modification[J]. Eur Heart J, 2013, 34(43): 3378-3388.
[20] OTSUKA F, BYRNE R A, YAHAGI K, et al.Neoatherosclerosis: overview of histopathologic findings and implications for intravascular imaging assessment[J]. Eur Heart J, 2015, 36(32): 2147-2159.
[21] LIU S, LI L, WANG H, et al.Recent Advances: From Cell Biology to Cell Therapy in Atherosclerosis Plaque via Stent Implantation[J]. Curr Med Chem, 2023, 30(31): 3582-3613.
[22] WHITTAKER A, MOORE J S, VASA-NICOTERA M, et al.Evidence for genetic regulation of endothelial progenitor cells and their role as biological markers of atherosclerotic susceptibility[J]. Eur Heart J, 2008, 29(3): 332-338.
[23] HE L, ZHANG C L, CHEN Q, et al.Endothelial shear stress signal transduction and atherogenesis: From mechanisms to therapeutics[J]. Pharmacol Ther, 2022, 235: 108152.
[24] CHIU J J, CHIEN S.Effects of disturbed flow on vascular endothelium: pathophysiological basis and clinical perspectives[J]. Physiol Rev, 2011, 91(1): 327-387.
[25] SHYY J Y, CHIEN S.Role of integrins in endothelial mechanosensing of shear stress[J]. Circ Res, 2002, 91(9): 769-775.
[26] MARTINET W, DE MEYER G R. Autophagy in atherosclerosis: a cell survival and death phenomenon with therapeutic potential[J]. Circ Res, 2009, 104(3): 304-317.
[27] LAVANDERO S, CHIONG M, ROTHERMEL B A, et al.Autophagy in cardiovascular biology[J]. J Clin Invest, 2015, 125(1): 55-64.
[28] LYU Z S, CAO X N, WEN Q, et al.Autophagy in endothelial cells regulates their haematopoiesis-supporting ability[J]. EBioMedicine, 2020, 53: 102677.
[29] HAN J, PAN X Y, XU Y, et al.Curcumin induces autophagy to protect vascular endothelial cell survival from oxidative stress damage[J]. Autophagy, 2012, 8(5): 812-825.
[30] ABDELLATIF M, SEDEJ S, CARMONA-GUTIERREZ D, et al.Autophagy in Cardiovascular Aging[J]. Circ Res, 2018, 123(7): 803-824.
[31] LIU H, HE Z, SIMON H U.Targeting autophagy as a potential therapeutic approach for melanoma therapy[J]. Semin Cancer Biol, 2013, 23(5): 352-360.

基金

深圳市南山区卫生科技计划项目“miR101-mTOR信号通路介导自噬在切应力调控内皮祖细胞治疗动物内皮损伤的机制研究”（2020007）; 深圳市南山区卫生科技计划项目“长链非编码RNA SNHG7 在缺血和再灌注损伤中的分子机制研究”（202034）; 深圳市三名工程资助院级项目“Max试验摄氧量在HFpEF、HFmrEF及HFrEF组患者中的诊断应用研究”（YJ20210007）