刘怡,陈敏琪,张翼,千忠吉

• 1:广东海洋大学化学与环境学院
• 2:广东海洋大学食品科技学院
• 3:广东海洋大学深圳研究院

摘要(Abstract)：

【目的】探究海洋土霉菌代谢产物epi-aszonalenin A(EAA)对氧化型低密度脂蛋白(ox-LDL)诱导HUVEC人脐静脉内皮细胞的动脉粥样硬化斑块内血管新生的作用。【方法】用CCK法检测细胞活力,DCFH-DA法测定活性氧(ROS)的含量,划痕实验检测细胞迁移能力,血管生成实验检测细胞成管能力,酶联免疫吸附法ELISA试剂盒检测LOX-1和VEGF蛋白表达情况,蛋白免疫印迹法检测MAPK通路、细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)的蛋白的表达情况,分子对接模拟EAA与LOX-1蛋白的相互作用。【结果】CCK法证明EAA对HUVEC细胞无明显毒性作用(P> 0.05);与空白组相比,EAA对HUVEC细胞的迁移和血管生成起到显著抑制作用(P <0.001);与对照组相比,随着实验组EAA浓度增加,细胞内ROS含量显著减少(P <0.001),LOX-1、VEGF、MAPK通路蛋白p38、JNK、ERK的磷酸化和ICAM-1、VCAM-1的表达也显著降低(P <0.001);此外EAA能与LOX-1形成稳定的相互作用。【结论】EAA能清除ROS,抑制HUVEC细胞炎性因子的表达和血管生成。

关键词(KeyWords)： 土曲霉;代谢产物;人脐静脉内皮细胞;血管生成;抗炎;氧化型低密度脂蛋白

Abstract:

Keywords:

基金项目(Foundation): 深圳市国际合作研究项目协同创新专项（GJHZ20190823111601682）;; 南海海洋生物医药资源研发公共服务平台项目（XM-202008-01B1）;; 广东省基础与应用基础研究基金（2020A1515011075）

作者(Author): 刘怡,陈敏琪,张翼,千忠吉

参考文献(References)：

• [1]LIBBY P.Atherosclerosis in[[
• [2]PIRILLO A,NORATA G D,CATAPANO A L.LOX
• [3]SAWAMURA T,KUME N,AOYAMA T,et al.An
• [4]AKHMEDOV A,ROZENBERG I,PANENI F,et al.420(6917):868Ox LDL,and atherosclerosis[J].Mediators of Inflammation,2013endothelial receptor for oxidized lo Nature,1997,386(6620):73Endothelial overexpression of LOX(24):152786.:868-874.:152786.74.]LIBBY P.Atherosclerosis in inflammation[J].Nature,2002,,1997,386(6620):73-77.inflammation[J].Nature,2002,77.l receptor for oxidized low w-density lipoprotein[J].-1 increases plaque-1,formation and promotes atherosclerosis in vivo[J].European Heart Journal,2014,35(40):2839-2848.
• [5]JUN J,REINKE C,BEDJA D,et al.Effect of intermittent hypoxia on atherosclerosis in apolipoprotein E-deficient mice[J].Atherosclerosis,2010,209(2):381-386.
• [6]FERRARA N,GERBER H P,LECOUTER J.The biology of VEGF and its receptors[J].Nature Medicine,2003,9(6):669-676.
• [7]AOKI K,WATANABE K,SATO M,et al.Effects of rhizoxin,a microbial angiogenesis inhibitor,on angiogenic endothelial cell functions[J].European Journal of Pharmacology,2003,459(2/3):131-138.
• [8]MORENO P R,PURUSHOTHAMAN K R,ZIAS E,et al.Neovascularization in human atherosclerosis[J].Current Molecular Medicine,2006,6(5):457-477.
• [9]PRADHAN R,SINGHVI G,DUBEY S K,et al.MAPKpathway:a potential target for the treatment of non-small-cell lung carcinoma[J].Future Medicinal Chemistry,2019,11(8):793-795.
• [10]毛洋,刘小琼,王洪梅,等.细胞间黏附分子1、血管细胞黏附分子1促进兔动脉粥样硬化斑块内血管新生[J].中国动脉硬化杂志,2014,22(3):217-222.
• [11]KOLODGIE F D,GOLD H K,BURKE A P,et al.Intraplaque hemorrhage and progression of coronary atheroma[J].The New England Journal of Medicine,2003,349(24):2316-2325.
• [12]PARMA L,BAGANHA F,QUAX P H A,et al.Plaque angiogenesis and intraplaque hemorrhage in atherosclerosis[J].European Journal of Pharmacology,2017,816:107-115.
• [13]MAUREA N,COPPOLA C,PISCOPO G,et al.Pathophysiology of cardiotoxicity from target therapy and angiogenesis inhibitors[J].Journal of Cardiovascular Medicine (Hagerstown,Md),2016,17(Suppl 1 Special issue on Cardiotoxicity from Antiblastic Drugs and Cardioprotection):e19-e26.
• [14]TOCCHETTI C G,GALLUCCI G,COPPOLA C,et al.The emerging issue of cardiac dysfunction induced by antineoplastic angiogenesis inhibitors[J].European Journal of Heart Failure,2013,15(5):482-489.
• [15]SCHUEFFLER A,ANKE T.Fungal natural products in research and development[J].Natural Product Reports,2014,31(10):1425-1448.
• [16]杨静明,杨文聪,刘亚月,等.化学诱导对一株海洋来源土曲霉C23-3次生代谢产物及其生物活性的影响[J].微生物学通报,2019,46(3):441-452.
• [17]ZHANG Y Y,ZHANG Y,YAO Y B,et al.Butyrolactone-Ifrom coral-derived fungus Aspergillus terreus attenuates neuro-inflammatory response via suppression of NF-κBpathway in BV-2 cells[J].Marine Drugs,2018,16(6):E202.
• [18]GUO F,LI Z L,XU X W,et al.Butenolide derivatives from the plant endophytic fungus Aspergillus terreus[J].Fitoterapia,2016,113:44-50.
• [19]RANK C,PHIPPS R K,HARRIS P,et al.Epi-Aszonalenins A,B,and C from Aspergillus novofumigatus[J].Tetrahedron Letters,2006,47(34):6099-6102.
• [20]MORRIS G M,HUEY R,LINDSTROM W,et al.Auto Dock4 and Auto Dock Tools4:Automated docking with selective receptor flexibility[J].Journal of Computational Chemistry,2009,30(16):2785-2791.
• [21]TROTT O,OLSON A J.Auto Dock Vina:improving the speed and accuracy of docking with a new scoring function,efficient optimization,and multithreading[J].Journal of Computational Chemistry,2010,31(2):455-461.
• [12]WANG T,BUTANY J.Pathogenesis of atherosclerosis[J].Diagnostic Histopathology,2017,23(11):473-478.
• [23]SAWAMURA T,KAKUTANI M,CHEN M Y,et al.LOX-1,an endothelial receptor for oxidized LDL Implications for induction of endothelium dysfunction in the pathogenesis of vascular diseases[C]//Lipoprotein Metabolism and Atherogenesis,2000:45:193-198.
• [24]KATTOOR A J,KANURI S H,MEHTA J L.Role of ox-LDL and LOX-1 in atherogenesis[J].Current Medicinal Chemistry,2019,26(9):1693-1700.
• [25]KATTOOR A J,POTHINENI N V K,PALAGIRI D,et al.Oxidative stress in atherosclerosis[J].Current Atherosclerosis Reports,2017,19(11):42.
• [26]HUBBARD A K,ROTHLEIN R.Intercellular adhesion molecule-1 (ICAM-1) expression and cell signaling cascades[J].Free Radical Biology and Medicine,2000,28(9):1379-1386.
• [27]MAO J J,LIU J T,PANG X M,et al.Nicotine induces the expression of C-reactive protein via MAPK-dependent signal pathway in U937 macrophages[J].Molecules and Cells,2012,34(5):457-461.