Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, 341000, China.
Department of Clinical Research Center of Vascular Abnormalities of Jiangxi Province, Ganzhou, 341000, China.
Sci Rep. 2024 Nov 18;14(1):28464. doi: 10.1038/s41598-024-80088-9.
The dibutyl phthalate (DBP) is a member of the phthalate family and is widely used as a plasticizer in daily life and production. However, the influence of DBP on the vascular developmental remains unclear.
In this study, we used zebrafish as a model organism to investigate the effects of DBP on vascular development in vivo. Death curves of zebrafish at different concentrations of DBP exposure and different times incubation were made firstly. Zebrafish embryos after fertilization for 5.5 h were exposed to different concentrations of DBP solution (0, 0.4, 0.8, 1.2 mg/L), the body length, yolk sac absorption area, mortality and heart rate of zebrafish were measured, and the number and area of sprouting of ventral vessels were quantified by transgenic fish system. Reactive oxygen species (ROS) in zebrafish embryos were observed by DCFH-DA staining. Super oxide dimutese (SOD) and catalase (CAT) were determined with ELISA kits.
We found that DBP increased the oxidative stress level of zebrafish exposed to DBP, and the genes related to vascular development also increased. Meanwhile, the activities of SOD and CAT were greatly decreased after DBP exposure. In the rescue experiment, we found that the antioxidant astaxanthin and the small molecule VEGF inhibitor ZM-306,416 can reverse the vascular dysplasia caused by DBP.
DBP induced vascular developmental toxicity by enhancing oxidative stress levels, activating HIF pathway, and interfering with the expression of vascular development-related pathways in zebrafish, results in the abnormal development of the subintestinal vessels in zebrafish.
邻苯二甲酸二丁酯（DBP）是邻苯二甲酸酯家族的一员，广泛用作日常生活和生产中的增塑剂。然而，DBP 对血管发育的影响尚不清楚。
本研究以斑马鱼为模型生物，体内研究 DBP 对血管发育的影响。首先制作不同浓度 DBP 暴露和不同孵育时间下斑马鱼的死亡曲线。将受精后 5.5 h 的斑马鱼胚胎暴露于不同浓度的 DBP 溶液（0、0.4、0.8、1.2 mg/L）中，测量斑马鱼的体长、卵黄囊吸收面积、死亡率和心率，并通过转基因鱼系统定量测量腹侧血管的发芽数量和面积。用 DCFH-DA 染色观察斑马鱼胚胎中的活性氧（ROS）。用 ELISA 试剂盒测定超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。
我们发现 DBP 增加了暴露于 DBP 的斑马鱼的氧化应激水平，与血管发育相关的基因也增加。同时，DBP 暴露后 SOD 和 CAT 的活性大大降低。在挽救实验中，我们发现抗氧化剂虾青素和小分子 VEGF 抑制剂 ZM-306,416 可以逆转 DBP 引起的血管发育不良。
DBP 通过增强氧化应激水平、激活 HIF 通路以及干扰斑马鱼血管发育相关通路的表达，引起血管发育毒性，导致斑马鱼的亚肠血管发育异常。
