The Key Laboratory of Modern Toxicology, Ministry of Education, Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, China; The Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing, China.
The Key Laboratory of Modern Toxicology, Ministry of Education, Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, China; The Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing, China; Safety Assessment and Research Center for Drug, Pesticide and Veterinary Drug of Jiangsu Province, Nanjing Medical University, Nanjing, China.
Toxicology. 2022 Jul;477:153279. doi: 10.1016/j.tox.2022.153279. Epub 2022 Aug 1.
Di-n-butyl phthalate (DBP) is ubiquitous in environment and has been detected in almost all human bodies. Few data could be found about the effects of DBP on cardiovascular system, though its reproductive toxicities have been studied extensively. This study aimed to explore effects of DBP on phenotypic switching of vascular smooth muscle cells (VSMCs), an essential step during the formation of atherosclerosis (AS). A7r5 cells were employed and exposed to various levels of DBP (10, 10, 10, 10, and 10 M) or DMSO as control. CCK-8 assay was used to detect the effects of DBP on cell viability. Expressions of mRNA/miRNAs and proteins were measured by qRT-PCR and western blotting, respectively. Bioinformatic analysis and dual-luciferase reporter assay were used to analyze the combination between miR-139-5p and Myocardin (MYOCD). Results revealed that DBP at 10 M prompted phenotypic switching from contractile to synthetic of VSMCs by inhibiting contractile VSMCs marker genes via suppressing the expression of MYOCD. Moreover, miR-139c-5p directly targeted MYOCD 3'UTR and modulated MYOCD expression. Besides, DBP inhibited the expression of MYOCD and VSMCs marker genes by upregulating miR-139-5p. Collectively, these data suggested that DBP could promote the phenotypic switching from contractile to synthetic of VSMCs in A7r5 cells through miR-139-5p-MYOCD.
邻苯二甲酸二正丁酯（DBP）在环境中无处不在，几乎存在于所有人的体内。虽然已经广泛研究了其生殖毒性，但关于 DBP 对心血管系统的影响的数据却很少。本研究旨在探讨 DBP 对血管平滑肌细胞（VSMCs）表型转换的影响，这是动脉粥样硬化（AS）形成过程中的一个重要步骤。本研究采用 A7r5 细胞，并暴露于不同浓度的 DBP（10、10、10、10 和 10 M）或 DMSO 作为对照。CCK-8 检测试剂盒用于检测 DBP 对细胞活力的影响。通过 qRT-PCR 和 Western blot 分别测量 mRNA/miRNA 和蛋白质的表达。生物信息学分析和双荧光素酶报告基因实验用于分析 miR-139-5p 和肌球蛋白重链结合蛋白 D（MYOCD）之间的结合。结果表明，DBP 在 10 M 浓度下通过抑制收缩型 VSMCs 标志物基因的表达，抑制肌球蛋白重链结合蛋白 D（MYOCD）的表达，从而促进 VSMCs 从收缩型向合成型表型转换。此外，miR-139-5p 直接靶向 MYOCD 3'UTR 并调节 MYOCD 表达。此外，DBP 通过上调 miR-139-5p 抑制 MYOCD 和 VSMCs 标志物基因的表达。综上所述，这些数据表明 DBP 可通过 miR-139-5p-MYOCD 促进 A7r5 细胞中 VSMCs 从收缩型向合成型表型转换。
