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Oxidative Medicine and Cellular Longevity Volume 2018 ,2018-11-25
Pyrrolidine Dithiocarbamate (PDTC) Inhibits DON-Induced Mitochondrial Dysfunction and Apoptosis via the NF-κB/iNOS Pathway
Research Article
Dan Wan 1 , 2 , 3 , 4 Qinghua Wu 5 , 6 Wei Qu 2 , 3 Gang Liu 1 Xu Wang 2 , 3 , 4
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DOI:10.1155/2018/1324173
Received 2018-07-06, accepted for publication 2018-10-16, Published 2018-10-16
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摘要

Oxidative stress is closely linked to the toxic responses of various cell types in normal and pathophysiological conditions. Deoxynivalenol (DON), an inducer of stress responses in the ribosome and the endoplasmic reticulum (ER), causes mitochondrial dysfunction and mitochondria-dependent apoptosis through oxidative stress in humans and animals. The NF-κB pathway, which is closely linked to oxidative stress, is hypothesized to be a critical signaling pathway for DON-induced toxicity and is a potential target for intervention. The present study was conducted to explore the protective effects of pyrrolidine dithiocarbamate (PDTC) from the toxic effects of DON in rat anterior pituitary GH3 cells. Our results showed that DON activated the NF-κB transcription factors and induced cellular oxidative stress, mitochondrial dysfunction, and apoptosis. Morphological studies using transmission electron microscopy (TEM) and cell apoptosis analyses suggested that PDTC prevented DON-induced mitochondrial dysfunction and apoptosis, probably by preventing the DON-induced translocation of NF-κB p65 into the nucleus, and by inhibiting DON-induced iNOS expression. This led to the blocking of the NF-κB pathway and inhibition of iNOS activity.

授权许可

Copyright © 2018 Dan Wan et al. 2018
This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

图表

DON-induced IκBα and NF-κB p65 phosphorylation in GH3 cells.

Nuclear translocation of phosphoryl-NF-κB p65 (p-p65 (Ser536)) induced by DON treatment (1200 ng/mL) and PDTC pretreatment (20 μM, 45 min) followed by DON treatment (1200 ng/mL) in GH3 cells, visualized through indirect immunofluorescence, using Alexa Fluor-conjugated secondary antibody. The nucleus was stained with PI. The panels show PI staining, Alexa Fluor staining, overlay, and the 3D plane of the cells. All photos were captured at 400x magnification. Phosphoryl-NF-κB p65 was upregulated and can be observed in the nucleus.

PDTC protects cells from DON-induced iNOS expression. (a) Cells were treated with DON, PDTC, and PDTC pretreatment followed by DON to assess iNOS transcription by qRT-PCR. (b). Cells were treated with DON, DON, PDTC, and PDTC pretreatment followed by DON to assess iNOS expression by western blotting. P values < 0.05 are indicated by a single asterisk, ∗. P values < 0.01 are indicated by double asterisks, ∗∗.

PDTC protects cells from DON-induced iNOS expression. (a) Cells were treated with DON, PDTC, and PDTC pretreatment followed by DON to assess iNOS transcription by qRT-PCR. (b). Cells were treated with DON, DON, PDTC, and PDTC pretreatment followed by DON to assess iNOS expression by western blotting. P values < 0.05 are indicated by a single asterisk, ∗. P values < 0.01 are indicated by double asterisks, ∗∗.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced mitochondrial injury in GH3 cells. Cells were treated with DON, PDTC, and PDTC pretreatment, followed by treatment with DON for 12 hours. (a, e) Cell treated with DON showing normal mitochondria. (b, f) Cell treated with PDTC showing normal mitochondria. (c, g) Cells with PDTC pretreatment, followed by treatment with DON for 12 hours showing normal mitochondria and tiny vacuoles.

PDTC protects cells from DON-induced apoptosis in GH3 cells. Cells in the control group (a), cells treated with PDTC (b) and DON (c), and cells pretreated with PDTC followed by DON treatment (d) were used to assess the apoptosis rate. Data are shown as means for three separate experiments performed in triplicate.

PDTC protects cells from DON-induced apoptosis in GH3 cells. Cells in the control group (a), cells treated with PDTC (b) and DON (c), and cells pretreated with PDTC followed by DON treatment (d) were used to assess the apoptosis rate. Data are shown as means for three separate experiments performed in triplicate.

PDTC protects cells from DON-induced apoptosis in GH3 cells. Cells in the control group (a), cells treated with PDTC (b) and DON (c), and cells pretreated with PDTC followed by DON treatment (d) were used to assess the apoptosis rate. Data are shown as means for three separate experiments performed in triplicate.

PDTC protects cells from DON-induced apoptosis in GH3 cells. Cells in the control group (a), cells treated with PDTC (b) and DON (c), and cells pretreated with PDTC followed by DON treatment (d) were used to assess the apoptosis rate. Data are shown as means for three separate experiments performed in triplicate.

A proposed mechanism of action for the protective effect of PDTC in DON-mediated mitochondrial dysfunction and apoptosis. DON indirectly activates the NF-κB signal pathway via the classical route of IκB/NF-κB p65 signaling. PDTC inhibits the translocation of NF-κB p65 and the transcription of iNOS, and thereby protects cells from mitochondrial dysfunction and cell apoptosis.

通讯作者

1. Gang Liu.Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic Process, Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Changsha, Hunan 410125, China, cas.cn.gangle.liu@gmail.com
2. Xu Wang.National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Wuhan 430070, China;MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Wuhan 430070, China;Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Wuhan 430070, China.wangxu@mail.hzau.edu.cn

推荐引用方式

Dan Wan,Qinghua Wu,Wei Qu,Gang Liu,Xu Wang. Pyrrolidine Dithiocarbamate (PDTC) Inhibits DON-Induced Mitochondrial Dysfunction and Apoptosis via the NF-κB/iNOS Pathway. Oxidative Medicine and Cellular Longevity ,Vol.2018(2018)

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