邻苯二甲酸二-2-乙基己酯通过ROS/PTEN/PI3K/AKT轴诱导HD11细胞凋亡和程序性坏死

doi:10.11843/j.issn.0366-6964.2023.12.033

Abstract

Abstract: To explore the mechanism of apoptosis and necroptosis induced by di(2-ethylhexyl)phthalate (DEHP) exposure through ROS/PTEN/PI3K/AKT pathway. HD11 cells were used in this study, and control group(C group), 30 μmol·L^-1 DEHP group(L group), 60 μmol·L^-1 DEHP group(M group), 90 μmol·L^-1 DEHP group(H group)were set. After cells were treated with different concentrations of DEHP for 24 h, cell survival was detected by CCK-8, oxidative stress was detected by biochemical kit, included reactive oxygen species (ROS)levels, malondialdehyde (MDA)content, total antioxidant capacity (T-AOC)levels, and total superoxide dismutase (T-SOD)and glutathione peroxidase (GSH-PX)activities. AO/EB staining and flow cytometry were used to detect the rate of apoptosis and necrosis, mRNA and protein expressions of PTEN/PI3K/AKT pathway genes and apoptosis and necroptosis related genes were detected by qRT-PCR and Western blot. The results showed that, compared with the control group, DEHP considerably reduced the viability of HD11 cells, with the median inhibitory concentration (IC₅₀)being 180.644 μmol·L^-1. Besides, compared with the control group, AO/EB staining and flow cytometry validated the L, M and H DEHP group with typical characteristics of apoptosis and necroptosis. Additionally, compared with the control group, ROS expression levels in groups L, M and H showed an increasing trend (P<0.01), and the content of MDA were considerably increased (P<0.01), the T-AOC level was significantly decreased (P<0.01), and the activities of T-SOD and GSH-PX were significantly decreased (P<0.01). It was suggested that DEHP exposure induced oxidative stress injury of HD11 cells. In addition, compared with the control group, DEHP exposure up-regulated the mRNA and protein expression levels of PTEN/Bax/Caspase-9/Caspase-3/RIPK1/RIPK3/MLKL and down-regulated the mRNA and protein expression levels of PI3K/AKT/BCL-2(P<0.01), these results suggested that DEHP exposure induced apoptosis and necroptosis of HD11 cells through the PTEN/PI3K/AKT signaling pathway. Overall, DEHP can induce apoptosis and necroptosis in HD11 cells by modulating the ROS/PTEN/PI3K/AKT axis, and there is a dose-effect manner.

Key words: DEHP, oxidative stress, apoptosis, necroptosis

CLC Number:

S859.8

LI Guangxing, CHEN Yang, CHEN Kaiting, WU Menglin, ZHANG Di, HUANG Xiaodan. Di (2-ethylhexyl) Phthalate Induces the Apoptosis and Necroptosis of HD11 Cells through the ROS/PTEN/PI3K/AKT Axis[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5240-5251.

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Di (2-ethylhexyl) Phthalate Induces the Apoptosis and Necroptosis of HD11 Cells through the ROS/PTEN/PI3K/AKT Axis

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