Institute of Cardiovascular Diseases, Chest Hospital, Tianjin University, Tianjin, 300222, China.
Department of Cardiology, Chest Hospital, Tianjin University, Tianjin, 300222, China.
Chemosphere. 2022 Dec;308(Pt 2):136383. doi: 10.1016/j.chemosphere.2022.136383. Epub 2022 Sep 8.
Phthalates are commonly used in variety of plastic products. Previously it has been revealed that di (2-ethylhexyl) phthalate (DEHP), as the most common member of the class of phthalates, may disturb cholesterol homeostasis and deregulate the inflammatory response, and leading to accelerate the atherosclerosis process. In this regard, the aim of the current study is to explore the underlying mechanism of DEHP-induced atherosclerosis through the increasing of foam cell formation and Vascular Smooth Muscle Cells (VSMCs) damage via the interaction of long-non coding RNA (GAS5) and miR-145-5p.
ApoE mice were used to evaluate the in vivo study. RAW264.7 and VSMCs were used to evaluate the effect of DEHP on formation of foam cell, cell proliferation, and cell damage in vitro. Animals were treated with DEHP (5% w/w of food) orally and cells were treated with medium containing of 100 μM DEHP; qRT-PCR, Western blotting, flowcytometry, IHC, oil red O, BODIPY, and autophagic vacuoles assay were used to evaluate the effect of DEHP on formation of atherosclerosis.
DEHP significantly accelerated the formation of atherosclerosis in mice and alter the lipid profile in mice. In addition, after treating VSMCs with DEHP, GAS5 was significantly up-regulated and miR-145-5p was down-regulated. In VSMCs treated with DEHP, we observed that GAS5 could be used as the competing endogenous RNA (ceRNA) of miR-145-5p to regulate the proliferation and apoptosis of VSMCs; and the expression of GAS5 was correlated with the expression of miR-145-5p. DEHP increased the ox-LDL uptake by macrophage and increasing the formation of foam cells. Besides, GAS5 knocking down reversed the effect of DEHP on foam cell formation and ox-LDL uptake.
DEHP could accelerate the atherosclerosis process through increasing VSMCs damage and formation of macrophage foam cell by increasing lipid uptake though down regulating lncRNA GAS5 and altering in regulation of miR-145-5p.
邻苯二甲酸酯广泛应用于各种塑料制品中。先前已经发现，作为邻苯二甲酸酯类最常见的成员之一，二（2-乙基己基）邻苯二甲酸酯（DEHP）可能会扰乱胆固醇的内稳态并使炎症反应失调，从而加速动脉粥样硬化过程。在这方面，本研究的目的是通过长链非编码 RNA（GAS5）与 miR-145-5p 的相互作用，通过增加泡沫细胞形成和血管平滑肌细胞（VSMCs）损伤，探讨 DEHP 诱导动脉粥样硬化的潜在机制。
使用载脂蛋白 E 基因敲除（ApoE-/-）小鼠进行体内研究。使用 RAW264.7 和 VSMCs 进行体外 DEHP 对泡沫细胞形成、细胞增殖和细胞损伤的影响。动物用 5%（w/w）的 DEHP 食物经口处理，细胞用含 100μM DEHP 的培养基处理；qRT-PCR、Western blot、流式细胞术、免疫组化、油红 O、BODIPY 和自噬小体检测用于评估 DEHP 对动脉粥样硬化形成的影响。
DEHP 可显著加速小鼠动脉粥样硬化的形成，并改变小鼠的脂质谱。此外，在 VSMCs 用 DEHP 处理后，GAS5 明显上调，miR-145-5p 下调。在用 DEHP 处理的 VSMCs 中，我们观察到 GAS5 可以作为 miR-145-5p 的竞争性内源性 RNA（ceRNA）来调节 VSMCs 的增殖和凋亡；并且 GAS5 的表达与 miR-145-5p 的表达相关。DEHP 通过增加巨噬细胞对氧化低密度脂蛋白（ox-LDL）的摄取并增加泡沫细胞的形成来加速动脉粥样硬化的发生。此外，GAS5 敲低可逆转 DEHP 对泡沫细胞形成和 ox-LDL 摄取的影响。
DEHP 通过下调 lncRNA GAS5 增加脂质摄取，并改变 miR-145-5p 的调控，增加巨噬细胞泡沫细胞的形成和 VSMCs 损伤，从而加速动脉粥样硬化进程。
