Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells

Excessive copper (Cu) has the potential risk to ecosystems and organism health, with its impact on dairy cow mammary glands being not well-defined. This study used a bovine mammary epithelial cell (MAC-T) model to explore how copper excess affects cellular oxidative stress, autophagy, ferroptosis, a...

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Main Authors: Ya Ting Fan, Dong Qiao Peng, Jing Lin Shen, Jun Hao Cui, Xin Yue Yang, Jin Ge Zhang, Yong Cheng Jin
Format: Article
Language:English
Published: Elsevier 2025-02-01
Series:Ecotoxicology and Environmental Safety
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Online Access:http://www.sciencedirect.com/science/article/pii/S0147651325001198
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author Ya Ting Fan
Dong Qiao Peng
Jing Lin Shen
Jun Hao Cui
Xin Yue Yang
Jin Ge Zhang
Yong Cheng Jin
author_facet Ya Ting Fan
Dong Qiao Peng
Jing Lin Shen
Jun Hao Cui
Xin Yue Yang
Jin Ge Zhang
Yong Cheng Jin
author_sort Ya Ting Fan
collection DOAJ
description Excessive copper (Cu) has the potential risk to ecosystems and organism health, with its impact on dairy cow mammary glands being not well-defined. This study used a bovine mammary epithelial cell (MAC-T) model to explore how copper excess affects cellular oxidative stress, autophagy, ferroptosis, and protein and lipid biosynthesis in milk. Results showed the increased intracellular ROS, MDA, and CAT (P < 0.05), alongside decreased T-SOD and GSH in CuSO4-treated cells (P < 0.05). Transmission electron microscopy and Ad-mCherry-GFP-LC3B assays revealed significant autophagosome accumulation in CuSO4 exposed cells (P < 0.05). Additionally, CuSO4 exposure modulated autophagy markers, evidenced by upregulation of genes such as LC3, ATG5, JNK1, and Beclin1, and downregulation of genes such as ATG4B, and p62 (P < 0.05). CuSO4 also led to notable mitochondrial changes, including size reduction, membrane rupture, and cristae loss, and reduced expression of the ferroptosis inhibitor GPX4 (P < 0.05). The expression of mTOR, HIF-1α and β-catenin signaling pathway were inhibited in differentiated MAC-T cells by CuSO4 exposure (P < 0.05), activated autophagy through activation of the AMPK-mTOR pathway which in turn affected downstream levels of genes related to milk protein and lipid. In conclusion, excessive copper induces oxidative stress in MAC-T cells, promoting autophagy through JNK-Bcl2, Beclin1-Vps34 and AMPK-mTOR pathways, leading to cell ferroptosis, as well as inhibits the cellular biosynthesis of milk protein and lipid.
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spelling doaj-art-3bb1a9721d3b4eb9bd23cbd25d1d5e3a2025-08-20T02:03:59ZengElsevierEcotoxicology and Environmental Safety0147-65132025-02-0129111778310.1016/j.ecoenv.2025.117783Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cellsYa Ting Fan0Dong Qiao Peng1Jing Lin Shen2Jun Hao Cui3Xin Yue Yang4Jin Ge Zhang5Yong Cheng Jin6College of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaCollege of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaCollege of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaCollege of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaCollege of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaCollege of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaCorresponding author.; College of Animal Science, Jilin University, Jilin Provincial Key Laboratory of Livestock and Poultry Feed and Feeding In Northeastern Frigid Area, Changchun 130062, ChinaExcessive copper (Cu) has the potential risk to ecosystems and organism health, with its impact on dairy cow mammary glands being not well-defined. This study used a bovine mammary epithelial cell (MAC-T) model to explore how copper excess affects cellular oxidative stress, autophagy, ferroptosis, and protein and lipid biosynthesis in milk. Results showed the increased intracellular ROS, MDA, and CAT (P < 0.05), alongside decreased T-SOD and GSH in CuSO4-treated cells (P < 0.05). Transmission electron microscopy and Ad-mCherry-GFP-LC3B assays revealed significant autophagosome accumulation in CuSO4 exposed cells (P < 0.05). Additionally, CuSO4 exposure modulated autophagy markers, evidenced by upregulation of genes such as LC3, ATG5, JNK1, and Beclin1, and downregulation of genes such as ATG4B, and p62 (P < 0.05). CuSO4 also led to notable mitochondrial changes, including size reduction, membrane rupture, and cristae loss, and reduced expression of the ferroptosis inhibitor GPX4 (P < 0.05). The expression of mTOR, HIF-1α and β-catenin signaling pathway were inhibited in differentiated MAC-T cells by CuSO4 exposure (P < 0.05), activated autophagy through activation of the AMPK-mTOR pathway which in turn affected downstream levels of genes related to milk protein and lipid. In conclusion, excessive copper induces oxidative stress in MAC-T cells, promoting autophagy through JNK-Bcl2, Beclin1-Vps34 and AMPK-mTOR pathways, leading to cell ferroptosis, as well as inhibits the cellular biosynthesis of milk protein and lipid.http://www.sciencedirect.com/science/article/pii/S0147651325001198AutophagyBovine Mammary Epithelial CellCopperFerroptosisMilk protein and lipid synthesis
spellingShingle Ya Ting Fan
Dong Qiao Peng
Jing Lin Shen
Jun Hao Cui
Xin Yue Yang
Jin Ge Zhang
Yong Cheng Jin
Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
Ecotoxicology and Environmental Safety
Autophagy
Bovine Mammary Epithelial Cell
Copper
Ferroptosis
Milk protein and lipid synthesis
title Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
title_full Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
title_fullStr Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
title_full_unstemmed Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
title_short Copper excess induces autophagy dysfunction and mitochondrial ROS-ferroptosis progression, inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
title_sort copper excess induces autophagy dysfunction and mitochondrial ros ferroptosis progression inhibits cellular biosynthesis of milk protein and lipid in bovine mammary epithelial cells
topic Autophagy
Bovine Mammary Epithelial Cell
Copper
Ferroptosis
Milk protein and lipid synthesis
url http://www.sciencedirect.com/science/article/pii/S0147651325001198
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