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| Developmental toxicity and molecular mechanism of Di-(2-ethylhexyl) phthalate exposure on zebrafish embryos |
| QIAN Xian1, FENG Weiwei1, MAO Guanghua 1, CHEN Yao 1, #br# LUO Mengna 1, WU Chaoqiong 1, YANG Liuqing 2, WU Xiangyang 1 |
| (1. School of Environment and Safety Engineering, 2. School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang Jiangsu 212013, China) |
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Abstract Objective： To study the effects of di-(2-ethylhexyl) phthalate (DEHP) on embryonic development of zebrafish and its potential molecular mechanism. Methods： Zebrafish embryos were randomly divided into 5 groups: blank control group, dimethyl sulfoxide group, and 8, 40, 200 μg/L DEHP groups, which were exposed to 96 hours post-fertilization (hpf). The mortality rate, hatching rate, malformation rate, heart rate, body length and the number of spontaneous movements of embryos were recorded. The activity of superoxide dismutase (SOD), glutathione catalase (GPx) and malondialdehyde (MDA) content in larvae were measured by enzymatic analysis. Real-time quantitative PCR was used to determine the expression of mRNA related to cell apoptosis, glucose transport, and the synthesis of proteins, fatty acids, and glycogen. Results： (1) Compared with the control group, the embryo/larvae mortality of zebrafish in 8 μg/L DEHP group was significantly increased (P＜0.01), and the larvae body length was greatly decreased (P＜0.05); in 40 and 200 μg/L DEHP groups, the mortality and malformation rate of zebrafish embryo/larvae were significantly increased (P＜0.01), and the hatching rate, heart rate, body length and the number of embryonic autonomous movement were greatly decreased (P＜0.05 or P＜0.01). (2) Compared with the control group, the activity of GPx in zebrafish larvae in 8 μg/L DEHP group was significantly decreased (P＜0.05); the activities of SOD and GPx in zebrafish larvae were significantly decreased and the content of MDA was greatly increased in 40 and 200 μg/L DEHP groups (P＜0.01). (3) Compared with the control group, the expression of Pi3k mRNA in glucose transport signaling pathway in 8 μg/L DEHP group was significantly up-regulated (P＜0.01); the mRNA expressions of Bax/Bcl-2, Caspase-3 and Ras were greatly up-regulated (P＜0.01), and the mRNA expressions of Akt and Pi3k in glucose transport signaling pathway were significantly up-regulated (P＜0.01), the mRNA expression of Erk1/2 was greatly down-regulated (P＜0.05 or P＜0.01), and the mRNA expressions of Mtor, Gsk-3 and GSK-3β in protein, fatty acid and glycogen synthesis signaling pathways were significantly up-regulated (P＜0.05 or P＜0.01) in 40 and 200 μg/L DEHP groups. Conclusion： DEHP may induce apoptosis, disrupt the expression of metabolism-related mRNA, cause oxidative damage, and thus affect the growth and development of zebrafish embryos, resulting in developmental toxicity.
［Key words］di-(2-ethylhexyl) phthalate; zebrafish embryo; developmental toxicity; mechanism of toxicity
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Received: 13 January 2023
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