Abstract
Copper (Cu) is one of the common heavy metal pollutants in the environment, and its toxic mechanisms have been extensively studied. However, the immunotoxicity induced by Cu remains rarely reported, and the effects of Cu on endoplasmic reticulum stress and mitochondria-mediated apoptosis have been little studied in the spleen. In this study, pigs were fed with different contents of Cu (10, 125, and 250 mg/kg Cu) for 80 days to establish a toxicity model. The results showed the Cu exposure triggered endoplasmic reticulum stress in the spleen, as evidenced by increased mRNA and protein levels of GRP94, GRP78, CHOP, XBP1, ATF6, and JNK; the positive rate of GRP78 increased by immunofluorescence analysis. Additionally, mitochondrial fission and fusion homeostasis were disrupted, the expression levels of mitochondrial dynamics–related genes Mfn1, Mfn2, and OPA1 decreased, DRP1 increased, and the positive rate of Mfn1 decreased by immunofluorescence analysis. Furthermore, Cu exposure could induce apoptosis, as demonstrated by the increased expression level of related proteins and genes Bak, Bax, Caspase-3, P53, and Cytc. In conclusion, these results suggest chronic Cu exposure can lead to endoplasmic reticulum stress and imbalance in mitochondrial dynamics and induced apoptosis of pig spleen, and these results provided new insights into the underlying mechanism of Cu exposure caused splenic toxicity, which has public health implications where humans and animals are exposed to copper contamination.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Chen H, Wang Y, Luo J, Kang M, Hou J, Tang R, Zhao L, Shi F, Ye G, He X et al (2022) Autophagy and apoptosis mediated nano-copper-induced testicular damage. Ecotoxicol Environ Saf 229:113039. https://doi.org/10.1016/j.ecoenv.2021.113039
Dai J, Yang X, Yuan Y, Jia Y, Liu G, Lin N, Xiao H, Zhang L, Chen J (2020) Toxicity, gut microbiota and metabolome effects after copper exposure during early life in SD rats. Toxicology 433–434:152395. https://doi.org/10.1016/j.tox.2020.152395
Duan X, Xu M, Zhou Y, Yan Z, Du Y, Zhang L, Zhang C, Bai L, Nie J, Chen G et al (2016) Effects of soil properties on copper toxicity to earthworm Eisenia fetida in 15 Chinese soils. Chemosphere 145:185–192. https://doi.org/10.1016/j.chemosphere.2015.11.099
Ge J, Huang Y, Lv M, Zhang C, Talukder M, Li J, Li J (2022) Cadmium induced Fak -mediated anoikis activation in kidney via nuclear receptors (AHR/CAR/PXR)-mediated xenobiotic detoxification pathway. J Inorg Biochem 227:111682. https://doi.org/10.1016/j.jinorgbio.2021.111682
Gorman GS, Chinnery PF, DiMauro S, Hirano M, Koga Y, McFarland R, Suomalainen A, Thorburn DR, Zeviani M, Turnbull DM (2016) Mitochondrial Dis Nat Rev Dis Primers 2:16080. https://doi.org/10.1038/nrdp.2016.80
Guo C, Wang J, Jing L, Ma R, Liu X, Gao L, Cao L, Duan J, Zhou X, Li Y et al (2018) Mitochondrial dysfunction, perturbations of mitochondrial dynamics and biogenesis involved in endothelial injury induced by silica nanoparticles. Environ Pollut 236:926–936. https://doi.org/10.1016/j.envpol.2017.10.060
Guo H, Wang Y, Cui H, Ouyang Y, Yang T, Liu C, Liu X, Zhu Y, Deng H (2021) Copper induces spleen damage through modulation of oxidative stress, apoptosis, DNA damage, and inflammation. Biol Trace Elem Res. https://doi.org/10.1007/s12011-021-02672-8
Igney FH, Krammer PH (2002) Death and anti-death: tumour resistance to apoptosis. Nat Rev Cancer 2(4):277–288. https://doi.org/10.1038/nrc776
Kumar V, Parihar RD, Sharma A, Bakshi P, Singh Sidhu GP, Bali AS, Karaouzas I, Bhardwaj R, Thukral AK, Gyasi-Agyei Y et al (2019) Global evaluation of heavy metal content in surface water bodies: a meta-analysis using heavy metal pollution indices and multivariate statistical analyses. Chemosphere 236:124364. https://doi.org/10.1016/j.chemosphere.2019.124364
Lei C, Liao J, Li Q, Shi J, Zhang H, Guo J, Han Q, Hu L, Li Y, Pan J et al (2021) Copper induces mitochondria-mediated apoptosis via AMPK-mTOR pathway in hypothalamus of pigs. Ecotoxicol Environ Saf 220:112395. https://doi.org/10.1016/j.ecoenv.2021.112395
Li JL, Li S, Tang ZX, Xu SW (2010) Oxidative stress-mediated cytotoxicity of cadmium in chicken splenic lymphocytes. Toxicol Lett 196:S122. https://doi.org/10.1016/j.toxlet.2010.03.429
Li L, Peng X, Guo L, Zhao Y, Cheng Q (2020) Sepsis causes heart injury through endoplasmic reticulum stress-mediated apoptosis signaling pathway. Int J Clin Exp Pathol 13(5):964–971
Liao J, Yang F, Chen H, Yu W, Han Q, Li Y, Hu L, Guo J, Pan J, Liang Z et al (2019) Effects of copper on oxidative stress and autophagy in hypothalamus of broilers. Ecotoxicol Environ Saf 185:109710. https://doi.org/10.1016/j.ecoenv.2019.109710
Liu H, Deng H, Cui H, Jian Z, Guo H, Fang J, Zuo Z, Deng J, Li Y, Wang X et al (2021a) Copper induces hepatocyte autophagy via the mammalian targets of the rapamycin signaling pathway in mice. Ecotoxicol Environ Saf 208:111656. https://doi.org/10.1016/j.ecoenv.2020.111656
Liu H, Lai W, Liu X, Yang H, Fang Y, Tian L, Li K, Nie H, Zhang W, Shi Y et al (2021b) Exposure to copper oxide nanoparticles triggers oxidative stress and endoplasmic reticulum (ER)-stress induced toxicology and apoptosis in male rat liver and BRL-3A cell. J Hazard Mater 401:123349. https://doi.org/10.1016/j.jhazmat.2020.123349
Mu M, Zhao H, Wang Y, Liu J, Fei D, Xing M (2019a) Arsenic trioxide or/and copper sulfate co-exposure induce glandular stomach of chicken injury via destruction of the mitochondrial dynamics and activation of apoptosis as well as autophagy. Ecotoxicol Environ Saf 185:109678. https://doi.org/10.1016/j.ecoenv.2019.109678
Mu M, Zhao H, Wang Y, Liu J, Fei D, Xing M (2019b) Arsenic trioxide or/and copper sulfate co-exposure induce glandular stomach of chicken injury via destruction of the mitochondrial dynamics and activation of apoptosis as well as autophagy. Ecotoxicol Environ Saf 185:109678. https://doi.org/10.1016/j.ecoenv.2019.109678
Orrenius S, Gogvadze V, Zhivotovsky B (2015) Calcium and mitochondria in the regulation of cell death. Biochem Biophys Res Commun 460(1):72–81. https://doi.org/10.1016/j.bbrc.2015.01.137
Renault TT, Manon S (2011) Bax: addressed to kill. Biochimie 93(9):1379–1391. https://doi.org/10.1016/j.biochi.2011.05.013
Song Y, Luo Z, Zhang L, Hogstrand C, Pan Y (2016) Endoplasmic reticulum stress and disturbed calcium homeostasis are involved in copper-induced alteration in hepatic lipid metabolism in yellow catfish Pelteobagrus fulvidraco. Chemosphere 144:2443–2453. https://doi.org/10.1016/j.chemosphere.2015.11.031
Thit A, Banta GT, Selck H (2015) Bioaccumulation, subcellular distribution and toxicity of sediment-associated copper in the ragworm Nereis diversicolor: the relative importance of aqueous copper, copper oxide nanoparticles and microparticles. Environ Pollut 202:50–57. https://doi.org/10.1016/j.envpol.2015.02.025
Wang X, Zhuang Y, Fang Y, Cao H, Zhang C, Xing C, Guo X, Li G, Liu P, Hu G et al (2021a) Endoplasmic reticulum stress aggravates copper-induced apoptosis via the PERK/ATF4/CHOP signaling pathway in duck renal tubular epithelial cells. Environ Pollut 272:115981. https://doi.org/10.1016/j.envpol.2020.115981
Wang Y, Zhao H, Mu M, Guo M, Xing M (2021b) Zinc offers splenic protection through suppressing PERK/IRE1-driven apoptosis pathway in common carp (Cyprinus carpio) under arsenic stress. Ecotoxicol Environ Saf 208:111473. https://doi.org/10.1016/j.ecoenv.2020.111473
Wang Y, Zhao H, Mu M, Guo M, Xing M (2021c) Zinc offers splenic protection through suppressing PERK/IRE1-driven apoptosis pathway in common carp (Cyprinus carpio) under arsenic stress. Ecotoxicol Environ Saf 208:111473. https://doi.org/10.1016/j.ecoenv.2020.111473
Zhang M, Du H, Huang Z, Zhang P, Yue Y, Wang W, Liu W, Zeng J, Ma J, Chen G et al (2018) Thymoquinone induces apoptosis in bladder cancer cell via endoplasmic reticulum stress-dependent mitochondrial pathway. Chem Biol Interact 292:65–75. https://doi.org/10.1016/j.cbi.2018.06.013
Zhao H, Wang Y, Fei D, Guo M, Yang X, Mu M, Yu H, Xing M (2019) Destruction of redox and mitochondrial dynamics co-contributes to programmed cell death in chicken kidney under arsenite or/and copper (II) exposure. Ecotoxicol Environ Saf 179:167–174. https://doi.org/10.1016/j.ecoenv.2019.04.062
Zhao Y, Cui LG, Talukder M, Cui JG, Zhang H, Li JL (2021) Lycopene prevents DEHP-induced testicular endoplasmic reticulum stress via regulating nuclear xenobiotic receptors and unfolded protein response in mice. Food Funct 12(24):12256–12264. https://doi.org/10.1039/d1fo02729h
Zheng X, Li S, Li J, Lv Y, Wang X, Wu P, Yang Q, Tang Y, Liu Y, Zhang Z (2020) Hexavalent chromium induces renal apoptosis and autophagy via disordering the balance of mitochondrial dynamics in rats. Ecotoxicol Environ Saf 204:111061. https://doi.org/10.1016/j.ecoenv.2020.111061
Zhu Y, Li X, Chen C, Wang F, Li J, Hu C, Li Y, Miao L (2012) Effects of aluminum trichloride on the trace elements and cytokines in the spleen of rats. Food Chem Toxicol 50(8):2911–2915. https://doi.org/10.1016/j.fct.2012.05.041
Funding
This work was supported by the National Natural Science Foundation of China (No. 32072930 and No. 31572585) and the National Key R&D Program of China (No. 2016YFD0501205 and No. 2017YFD0502200).
Author information
Authors and Affiliations
Contributions
Kai Zhang did the data analysis and wrote the manuscript. Experimental design, material preparation, and data collection were completed by Kai Zhang, Jianzhao Liao, Zhuoying Hu, and Quanwei Li. The manuscript was revised by Dr. Jianzhao Liao. Manuscripts were reviewed by Lianmei Hu, Jianying Guo, Ying Li, Hui Zhan, and Jiaqiang Pan. Professor Zhaoxin Tang directed the research. All authors have read and approved the submitted version. Kai Zhang and Jianzhao Liao made equal contributions to this work.
Corresponding author
Ethics declarations
Ethics approval
The procedures of animal experiments were authorized and performed by the Ethics Committee of South China Agricultural University (approval number: No. 2019A015).
Consent to participate
Not applicable.
Consent for publication
All the authors have consented for the publication of this research.
Conflict of interest
The authors declare no competing interests.
Additional information
Responsible Editor: Mohamed M. Abdel-Daim
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhang, K., Liao, J., Hu, Z. et al. Mitochondria-mediated apoptosis and endoplasmic reticulum stress are involved in the toxicity induced by copper in the porcine spleen. Environ Sci Pollut Res 30, 94928–94939 (2023). https://doi.org/10.1007/s11356-023-28621-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11356-023-28621-7