Department of Public Health, Faculty of Life Sciences, Kumamoto University School of Medicine, Kumamoto, Japan; Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan.
Am J Med Sci. 2019 Jun;357(6):492-506. doi: 10.1016/j.amjms.2019.02.012. Epub 2019 Feb 14.
Plasminogen activator inhibitor type 1 promotes formation of endothelial microparticles with procoagulant activity. However, it remains unclear whether di-(2-ethylhexyl) phthalate, a peroxisome proliferator-activated receptor α agonist, influences microparticle formation.
The effect of di-(2-ethylhexyl) phthalate on release of tissue factor-bearing microparticles was investigated using human M1 macrophages.
Exposure of M1 macrophages to di-(2-ethylhexyl) phthalate significantly upregulated expression of plasminogen activator inhibitor type 1, whereas incubation of macrophages with small interfering RNA for peroxisome proliferator-activated receptor α attenuated it. Di-(2-ethylhexyl) phthalate significantly increased the tissue factor protein level in culture supernatants of M1 macrophages, but not M2 macrophages. After purification of proteins by centrifugal filtration, western blotting detected 2 high molecular weight bands of tissue factor-bearing microparticles in culture supernatants of M1 macrophages. The upper band showed binding to factor VIIa and tissue factor pathway inhibitor, unlike the lower band. This suggested heterogeneity of the procoagulant activity of tissue factor-bearing microparticles, presumably dependent upon encryption/decryption of tissue factor. Phosphatidylserine contributes to tissue factor decryption, and western blotting revealed that the density of phosphatidylserine was reduced in the upper tissue factor band compared with the lower band. Di-(2-ethylhexyl) phthalate also upregulated transforming growth factor-β1 protein production by M1 macrophages. Moreover, silencing of Smad2, Smad3 or Smad4 attenuated plasminogen activator inhibitor type 1 expression and tissue factor-release from macrophages after di-(2-ethylhexyl) phthalate stimulation.
Di-(2-ethylhexyl) phthalate promotes formation of tissue factor-bearing microparticles in human M1 macrophages via the transforming growth factor-β1/Smad/ plasminogen activator inhibitor type 1 signaling pathway.
纤溶酶原激活物抑制剂 1 促进具有促凝活性的内皮微粒的形成。然而，二-(2-乙基己基)邻苯二甲酸酯（过氧化物酶体增殖物激活受体α激动剂）是否影响微粒形成仍不清楚。
使用人 M1 巨噬细胞研究二-(2-乙基己基)邻苯二甲酸酯对组织因子携带微粒释放的影响。
二-(2-乙基己基)邻苯二甲酸酯暴露于 M1 巨噬细胞显著上调纤溶酶原激活物抑制剂 1 的表达，而巨噬细胞孵育过氧化物酶体增殖物激活受体α 的小干扰 RNA 则减弱了它。二-(2-乙基己基)邻苯二甲酸酯显著增加了 M1 巨噬细胞培养上清液中的组织因子蛋白水平，但对 M2 巨噬细胞没有影响。用离心过滤法纯化蛋白质后，Western blot 在 M1 巨噬细胞培养上清液中检测到 2 个组织因子携带微粒的高分子量条带。上带与因子 VIIa 和组织因子途径抑制剂结合，与下带不同。这表明组织因子携带微粒的促凝活性存在异质性，可能依赖于组织因子的加密/解密。磷脂酰丝氨酸有助于组织因子解密，Western blot 显示与下带相比，上带的磷脂酰丝氨酸密度降低。二-(2-乙基己基)邻苯二甲酸酯还上调了 M1 巨噬细胞转化生长因子-β1 蛋白的产生。此外，沉默 Smad2、Smad3 或 Smad4 可减弱二-(2-乙基己基)邻苯二甲酸酯刺激后巨噬细胞中纤溶酶原激活物抑制剂 1 的表达和组织因子的释放。
二-(2-乙基己基)邻苯二甲酸酯通过转化生长因子-β1/Smad/纤溶酶原激活物抑制剂 1 信号通路促进人 M1 巨噬细胞中组织因子携带微粒的形成。
