Page 15 - 南京医科大学学报自然科学版

第42卷第11期 南京医科大学学报（自然科学版）
2022年11月 Journal of Nanjing Medical University（Natural Sciences） ·1507 ·

·基础研究·

邻苯二甲酸二（2⁃乙基己基）酯通过抑制PI3K/AKT信号通路损

害人冠状动脉内皮细胞的功能

孙立富，季宇萌，王旭枫，余冬敏，李 奔，路 鹏，王晓伟 *

南京医科大学第一附属医院心脏大血管外科，江苏 南京 210029

［摘 要］ 目的：探究邻苯二甲酸二（2⁃乙基己基）酯［di（2⁃ethylhexyl）phthalate，DEHP］暴露对PI3K/AKT信号通路和人冠状动
脉内皮细胞（human coronary artery endothelial cell，HCAEC）的影响，探索DEHP暴露致HCAEC损伤的机制。方法：根据相关文
献研究，用不同浓度DEHP处理HCAEC，设置对照组（DMSO）和实验组（DEHP 128、256、384、512 μmol/L）。用CCK⁃8法测定细
胞增殖能力；体外成管实验评估细胞成管能力；流式细胞术评估细胞凋亡水平；Western blot 检测细胞中 p⁃PI3K、
PI3K、p⁃AKT、AKT和凋亡相关蛋白BAX、Bcl⁃2、cleaved Caspase⁃3、Caspase⁃3的水平。结果：CCK⁃8结果显示，DEHP以浓度和
时间依赖的方式抑制HCAEC的增殖（P < 0.05）。体外成管实验结果显示，实验组细胞形态多呈孤立的圆形、卵圆形，当DEHP
浓度≥256 μmol/L时，其形成交点数目较对照组减少（P < 0.05）。流式细胞实验结果显示，实验组凋亡比例较对照组上升（P <
0.05）。Western blot 结果显示实验组 Bcl⁃2 表达水平较对照组降低，同时 BAX 和 cleaved Caspase⁃3/Caspase⁃3 的水平升高（P <
0.05）；当DEHP≥256 μmol/L时，p⁃PI3K/PI3K、p⁃AKT/AKT比值较对照组降低（P < 0.05）。结论：DEHP可通过抑制PI3K/AKT/信
号通路诱导HCAEC凋亡，抑制其增殖、血管形成能力。
［关键词］ 邻苯二甲酸二（2⁃乙基己基）酯；人冠状动脉内皮细胞；凋亡；PI3K/AKT
［中图分类号］ R329.28 ［文献标志码］ A ［文章编号］ 1007⁃4368（2022）11⁃1507⁃08
doi：10.7655/NYDXBNS20221102

Di ⁃（2 ⁃ ethylhexyl）phthalate impairs the functions of human coronary artery endothelial
cells by inhibiting PI3K/AKT signal pathway

SUN Lifu，JI Yumeng，WANG Xufeng，YU Dongmin，LI Ben，LU Peng，WANG Xiaowei *
Department of Cardiovascular Surgery，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，
China

［Abstract］ Objective：This study aims to investigate the effects of di（2⁃ethylhexyl）phthalate（DEHP）exposure on PI3K/AKT signal
pathway and human coronary artery endothelial cells（HCAECs），and to provide a theoretical basis for the treatment of HCAECs
damaged by DEHP exposure. Methods：According to relevant literature research，HCAECs were treated with different concentrations
of DEHP and divided into control group（DMSO）and experimental group（DEHP 128，256，384，512 μmol/L）. The ability of cell
proliferation was measured by CCK⁃ 8 assay，the ability of cell tube formation was evaluated by in vitro tube formation assay，the level
of apoptosis was evaluated by flow cytometry，and the levels of p⁃PI3K，PI3K，p⁃AKT，AKT and apoptosis⁃related proteins BAX，Bcl⁃2，
cleaved Caspase⁃3 and Caspase⁃3 were detected by Western blot. Results：CCK⁃8 assay showed that DEHP inhibited the proliferation
ability of HCAECs in a concentration⁃ and time⁃dependent manner（P < 0.05）. The results of in vitro tube formation assay showed that
the morphology of cells in the experimental group was mostly isolated round and oval. The number of intersections formed was reduced
compared with the control group（P < 0.05）when the DEHP concentration was ≥256 μmol/L. The results of flow cytometry showed that
the proportion of apoptosis in the experimental group was higher than that in the control group（P < 0.05）. Western blot results showed
that the expression level of Bcl⁃2 in the experimental group was lower than that in the control group，while the levels of BAX and

［基金项目］ 国家自然科学基金（81773445，81573234）；江苏省“333”工程项目（LGY2016006）；江苏省卫生健康委医学科研项
目（ZDA2020004）
∗
通信作者（Corresponding author），E⁃mail：wangxiaowei@njmu.edu.cn