海洋土曲霉C23-3代谢产物epi-aszonalenin A对内皮细胞的活性机制

【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。

Activie Mechanism of Epi-aszonalenin A Derived from Aspergillus terreus C23-3 on Endothelial Cells

【Objective】To analyze the effect of epi-aszonalenin A(EAA),a metabolite of coral-derived fungus Aspergillus terreus,on angiogenesis in atherosclerotic plaques induced by oxidized low-density lipoprotein(ox-LDL).【Methods】CCK method was used to detect cell viability;DCFH-DA was used to detect ROS content;scratch test was used to analyze the migration ability of human umbilical vein endothelial cells(HUVEC);angiogenesis was used to detect tube capacity;ELISA kit was used to detect the LOX-1 and VEGF proteins;western blotting was used to detect the expression of MAPK pathway,ICAM-1,VCAM-1 proteins;Molecular docking was used to simulate the interaction of EAA with LOX-1 protein.【Results】CCK method showed that EAA had no significant toxic effect on HUVEC(P>0.05).Compared with the blank control,EAA significantly inhibited cell migration and angiogenesis.Compared with the control group,ETT concentration in the experimental group increased.ROS content decreased significantly(P<0.001).LOX-1 and VEGF the expression of phosphorylation of p38,JNK,and ERK in MAPK pathway and ICAM-1 and VCAM-1 decreased gradually(P<0.001).Moreover,it can form a stable interaction with LOX-1.【Conclusion】EAA can clear ROS and inhibit the expression of inflammatory factors and angiogenesis in HUVEC.