Page 27 - 南京医科大学学报自然科学版

第43卷第2期 南京医科大学学报（自然科学版）
2023年2月 Journal of Nanjing Medical University（Natural Sciences） ·169 ·

·基础医学·

DNA 甲基化调控 p16 表达在替格瑞洛改善血管内皮功能中的

作用及机制

朱伯谦，宋兵战，陈 凯，姜 旭 ，王振兴 *
*
南京中医药大学附属医院心内科，江苏 南京 210029

［摘 要］ 目的：探讨替格瑞洛对氧化型低密度脂蛋白（oxidized low⁃density lipoprotein，ox⁃LDL）诱导内皮损伤的影响及机
制。方法：采用ELISA法检测人血清内皮素⁃1、一氧化氮含量，CCK⁃8法、EdU法、Transwell法检测细胞活力、增殖及迁移能力，
流式细胞术检测细胞凋亡，real⁃time PCR 法、Western blot 法检测 DNA 甲基化转移酶 1（DNA Methyltransferase 1，DNMT1）、p16
表达，慢病毒过表达载体构建p16的过表达系统，慢病毒敲除载体构建DNMT1和p16的敲除系统，甲基化特异性PCR法测定
p16启动子区甲基化水平。结果：替格瑞洛促进ox⁃LDL诱导的人脐静脉内皮细胞（human umbilical endothelial cell，HUVEC）活
力、增殖及迁移能力，并显著抑制ox⁃LDL诱导的细胞凋亡；替格瑞洛抑制ox⁃LDL诱导的HUVEC中p16表达，而过表达p16可逆
转替格瑞洛对ox⁃LDL 诱导HUVEC 损伤的保护作用；DNMT1通过影响p16启动子区的甲基化水平抑制p16表达。结论：替格
瑞洛可通过DNMT1介导的DNA甲基化调控p16表达，进而减轻ox⁃LDL诱导的HUVEC损伤，改善血管内皮功能。
［关键词］ 替格瑞洛；氧化型低密度脂蛋白；人脐静脉内皮细胞；p16；DNA甲基化转移酶1
［中图分类号］ R329.25 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）02⁃169⁃10
doi：10.7655/NYDXBNS20230204

The role and mechanism of ticagrelor on the vascular endothelial function through
DNMT1/p16 signaling pathwuay

*
ZHU Boqian，SONG Bingzhan，CHEN Kai，JIANG Xu ，WANG Zhenxing *
Department of Cardiology，Affiliated Hospital of Nanjing University of Chinese Medicine，Nanjing，210029，China

［Abstract］ Objective：To investigate the effect and mechanism of ticagrelor on endothelial injury induced by ox⁃LDL. Methods：
ELISA method was used to detect the serum ET⁃1 and NO contents. Cell viability，proliferation and migration ability were determined
by CCK⁃8，EdU kit and Transwell assay respectively. Annexin V⁃PI assay was used to detect cell apoptosis. The levels of DNMT1 and
p16 were detected by Real⁃time PCR and Western blot. The lentiviral overexpression vector pLVX⁃puro⁃EGFP was used to construct
the p16 overexpression system. The lentiviral knockout vector pLKO.1 was used to construct the DNMT1 and p16 knockout system.
The methylation status of p16 promoter region was determined by the methylation⁃specific PCR method. Results：Ticagrelor promoted
the viability，proliferation and migration ability of human umbilical endothelial cell（HUVEC）treated with ox⁃LDL，and significantly
inhibited ox ⁃ LDL ⁃ induced apoptosis. Ticagrelor down ⁃ regulated the expression of p16 in HUVECs induced by ox ⁃ LDL，and
overexpressed p16 could reverse the protective effect of ticagrelor on HUVECs induced by ox⁃LDL. DNMT1 inhibited the expression of
p16 by affecting the methylation level of p16 promoter region. Conclusion：Ticagrelor could reduce the injury of HUVECs induced by
ox⁃LDL and improve endothelial function. The mechanism might be related to the regulation of DNMT1/p16 signaling pathway.
［Key words］ ticagrelor；ox⁃LDL；human umbilical vein endothelial cells；p16；DNMT1
［J Nanjing Med Univ，2023，43（02）：169⁃178］

［基金项目］ 白求恩·医学科学研究基金项目（AX084ES）；国家自然科学基金青年项目（81900237）；江苏省自然科学基金青年
项目（BK20191093）
∗
通信作者（Corresponding author），E⁃mail：15996225138@163.com；zxwang518@sina.com