Cargando…

Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency

Sevoflurane (Sev) is one of the commonly used inhalation anesthetic chemicals in clinics. It has great impact on spermatogenesis and fertilization in male animals. The underlying mechanism remains largely unexplored. Based on our previous research, we hypothesized that Sev induced iron metabolism di...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Xue, Zuo, Yong, Zhang, Jianhua, Zhang, Di, Naeem, Muhammad, Chang, Yanzhong, Shi, Zhenhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10279888/
https://www.ncbi.nlm.nih.gov/pubmed/37346174
http://dx.doi.org/10.3389/fcell.2023.1184632
_version_ 1785060686014971904
author Zhang, Xue
Zuo, Yong
Zhang, Jianhua
Zhang, Di
Naeem, Muhammad
Chang, Yanzhong
Shi, Zhenhua
author_facet Zhang, Xue
Zuo, Yong
Zhang, Jianhua
Zhang, Di
Naeem, Muhammad
Chang, Yanzhong
Shi, Zhenhua
author_sort Zhang, Xue
collection PubMed
description Sevoflurane (Sev) is one of the commonly used inhalation anesthetic chemicals in clinics. It has great impact on spermatogenesis and fertilization in male animals. The underlying mechanism remains largely unexplored. Based on our previous research, we hypothesized that Sev induced iron metabolism disturbance in the testis and epididymis and inhibited the spermatogenesis. In this study, two-month-old C57BL/6 male mice were treated with 3% Sev for 6 h, and their fertility (including sperm concentration, sperm mobility, and the number of offspring) was evaluated. Mice testis, epididymis, and sperm were harvested and subjected to Western blot analysis and immunofluorescence analysis. Iron levels were reflected by the gene expression of iron metabolism-related proteins (including ferritin, TfR1, and FpN1) and ICP-MS and Perl’s iron staining. Electron transport and oxidative phosphorylation levels were measured by Oxygraph-2k and ATP contents. The activity of ribonucleotide reductase was evaluated by assay kit. DNA synthesis status in testis and/or epididymis was marked with BrdU. Cell proliferation was evaluated by double immunofluorescence staining of specific protein marker expression. Our results revealed that the mice exposed to Sev showed damaged testicular and epididymis structure and significantly reduced the sperm concentration, sperm motility, and fertility. Sev decreases the iron levels through down-regulating the expression of H-ferritin, L-ferritin, and FpN1, and up-regulating the expression of TfR1 in the testis and epididymis. Iron levels also significantly reduced in germ cells which decrease the number of germ cells, including sperm, Sertoli cells, and primary spermatocyte. Iron deficiency not only decreases electron transport, oxidative phosphorylation level, and ATP production but also suppresses the activity of ribonucleotide reductase and the expression of Ki67, DDX4, GATA1, and SCP3, indicating that Sev affects the spermatogenesis and development. Meanwhile, Sev impaired the blood-testis barrier by decreasing the ZO1 expression in the testis and epididymis. The damage effect induced by Sev can be significantly ameliorated by iron supplementation. In conclusion, our study illustrates a new mechanism by which Sev inhibits spermatogenesis and fertility through an oxidative phosphorylation pathway due to iron deficiency of epididymis and testis or sperm. Furthermore, the damaging effects could be ameliorated by iron supplementation.
format Online
Article
Text
id pubmed-10279888
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-102798882023-06-21 Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency Zhang, Xue Zuo, Yong Zhang, Jianhua Zhang, Di Naeem, Muhammad Chang, Yanzhong Shi, Zhenhua Front Cell Dev Biol Cell and Developmental Biology Sevoflurane (Sev) is one of the commonly used inhalation anesthetic chemicals in clinics. It has great impact on spermatogenesis and fertilization in male animals. The underlying mechanism remains largely unexplored. Based on our previous research, we hypothesized that Sev induced iron metabolism disturbance in the testis and epididymis and inhibited the spermatogenesis. In this study, two-month-old C57BL/6 male mice were treated with 3% Sev for 6 h, and their fertility (including sperm concentration, sperm mobility, and the number of offspring) was evaluated. Mice testis, epididymis, and sperm were harvested and subjected to Western blot analysis and immunofluorescence analysis. Iron levels were reflected by the gene expression of iron metabolism-related proteins (including ferritin, TfR1, and FpN1) and ICP-MS and Perl’s iron staining. Electron transport and oxidative phosphorylation levels were measured by Oxygraph-2k and ATP contents. The activity of ribonucleotide reductase was evaluated by assay kit. DNA synthesis status in testis and/or epididymis was marked with BrdU. Cell proliferation was evaluated by double immunofluorescence staining of specific protein marker expression. Our results revealed that the mice exposed to Sev showed damaged testicular and epididymis structure and significantly reduced the sperm concentration, sperm motility, and fertility. Sev decreases the iron levels through down-regulating the expression of H-ferritin, L-ferritin, and FpN1, and up-regulating the expression of TfR1 in the testis and epididymis. Iron levels also significantly reduced in germ cells which decrease the number of germ cells, including sperm, Sertoli cells, and primary spermatocyte. Iron deficiency not only decreases electron transport, oxidative phosphorylation level, and ATP production but also suppresses the activity of ribonucleotide reductase and the expression of Ki67, DDX4, GATA1, and SCP3, indicating that Sev affects the spermatogenesis and development. Meanwhile, Sev impaired the blood-testis barrier by decreasing the ZO1 expression in the testis and epididymis. The damage effect induced by Sev can be significantly ameliorated by iron supplementation. In conclusion, our study illustrates a new mechanism by which Sev inhibits spermatogenesis and fertility through an oxidative phosphorylation pathway due to iron deficiency of epididymis and testis or sperm. Furthermore, the damaging effects could be ameliorated by iron supplementation. Frontiers Media S.A. 2023-06-06 /pmc/articles/PMC10279888/ /pubmed/37346174 http://dx.doi.org/10.3389/fcell.2023.1184632 Text en Copyright © 2023 Zhang, Zuo, Zhang, Zhang, Naeem, Chang and Shi. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Zhang, Xue
Zuo, Yong
Zhang, Jianhua
Zhang, Di
Naeem, Muhammad
Chang, Yanzhong
Shi, Zhenhua
Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
title Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
title_full Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
title_fullStr Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
title_full_unstemmed Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
title_short Sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
title_sort sevoflurane inhibited reproductive function in male mice by reducing oxidative phosphorylation through inducing iron deficiency
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10279888/
https://www.ncbi.nlm.nih.gov/pubmed/37346174
http://dx.doi.org/10.3389/fcell.2023.1184632
work_keys_str_mv AT zhangxue sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency
AT zuoyong sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency
AT zhangjianhua sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency
AT zhangdi sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency
AT naeemmuhammad sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency
AT changyanzhong sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency
AT shizhenhua sevofluraneinhibitedreproductivefunctioninmalemicebyreducingoxidativephosphorylationthroughinducingirondeficiency