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Zinc Protects against Heat Stress–Induced Apoptosis via the Inhibition of Endoplasmic Reticulum Stress in TM3 Leydig Cells

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Abstract

Heat stress (HS)-induced apoptosis in Leydig cells is mediated by various molecular mechanisms, including endoplasmic reticulum (ER) stress. Zinc, an inorganic mineral element, exhibits several cytoprotective properties, but its potential protective action against Leydig cell apoptosis and the related molecular mechanisms has not been fully elucidated. In this study, we evaluated the effects of zinc sulfate, a predominant chemical form of zinc, exerted on cell viability, apoptosis, and testosterone production in HS-treated TM3 Leydig cells and investigated the underlying signaling pathways. HS treatment inhibited cell viability and induced apoptosis, which was accompanied by the induction of the activity of caspase 3, an executioner of apoptosis, involved in the expression of pro-apoptotic protein B cell lymphoma 2-associated X protein (Bax), and in the reduction of the expression of anti-apoptotic protein B cell lymphoma 2 (Bcl-2), thereby activating ER stress marker protein expression (glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP)). However, zinc sulfate led to the attenuation of deleterious effects, including increases in apoptosis, caspase-3 activity, Bax, GRP78, and CHOP expression, and decreases in cell viability and Bcl-2 protein expression in cells treated with HS or thapsigargin (an ER stress activator). Furthermore, 4-phenylbutyric acid (an ER stress inhibitor) treatment markedly alleviated the HS-induced adverse effects in cells exposed to HS, which was similar to zinc sulfate. Additionally, zinc sulfate supplementation in the culture medium effectively restored the HS-induced decrease in testosterone levels in HS-treated cells. In summary, these findings indicate that HS triggers apoptosis in TM3 Leydig cells via the ER stress pathway and that zinc confers protection against these detrimental effects. This study provides new insights into the benefits of using zinc against HS-induced apoptosis and cell injury.

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Abbreviations

Zn:

Zinc

HS:

Heat stress

ER:

Endoplasmic reticulum

Bax:

B cell lymphoma 2-associated X protein

Bcl-2:

B cell lymphoma 2

GRP78:

Glucose-regulated protein 78

CHOP:

CCAAT/enhancer-binding protein (C/EBP) homologous protein

HSP70:

Heat shock protein 70

4-PBA:

4-Phenylbutyrate

Tg:

Thapsigargin

3β-HSD:

3β-hydroxysteroid dehydrogenase

StAR:

Steroidogenic acute regulatory protein

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Acknowledgements

We thank Editage (Cactus Communications) for the revision of English grammar.

Funding

This research was funded by the Start-up Research Foundation for Newly Introduced Doctor of Anhui Science & Technology University (No. DKYJ201801), Anhui Natural Science Foundation (No. 1908085QC145), National Natural Science Foundation of China (31702306), Natural Science Key Project of Education Department of Anhui (No. KJ2020A0082), Project for Outstanding Youth of College in Anhui (gxyq2020038), and Collaborative Innovation Project of Anhui College (GXXT-2019-035).

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  1. College of Animal Science, Anhui Science and Technology University, Fengyang, 233100, Anhui, China

    Yongjie Xiong, Jing Li & Shaojun He

  2. Anhui Province Key Laboratory of Animal Nutritional Regulation and Health, Anhui Science and Technology University, Fengyang, 233100, Anhui, China

    Yongjie Xiong, Jing Li & Shaojun He

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  1. Yongjie Xiong
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  2. Jing Li
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Correspondence to Shaojun He.

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Xiong, Y., Li, J. & He, S. Zinc Protects against Heat Stress–Induced Apoptosis via the Inhibition of Endoplasmic Reticulum Stress in TM3 Leydig Cells. Biol Trace Elem Res 200, 728–739 (2022). https://doi.org/10.1007/s12011-021-02673-7

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