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Depletion of serum-derived exosomes aggravates heat stress-induced damage of bovine mammary epithelial cells

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Abstract

Background

Exosomes are involved in intercellular communication, affecting many physiological and pathological process. The present study evaluated the effects of serum exosomes on the function of bovine mammary epithelial cells (BMECs) and milk synthesis under heat stress.

Methods and results

We cultured the BMECs in fetal bovine serum (FBS) or exosome-free FBS medium and examined, their viability using CCK-8 kit. The results showed that culturing the cells in an exosome-free medium decreased viability and increased the levels of reactive oxygen species. The BMECs cultured in the exosome-free medium had reduced mitochondrial membrane potential, decreased manganese superoxide dismutase activity, and disrupted mitochondrial dynamics. They exhibited apoptosis due to upregulated Drp1, Fis1, Bax and HSP70. Lastly, we observed downregulation of milk fat and lactoprotein-related genes: mTOR, PPARγ, p-mTOR and ADD1 and SREBP1, ELF5, and CSN2, respectively, after culturing the cells in an exosome-free medium. These negative effects of the exosome-free medium on the BMECs could be further reinforced under heat stress.

Conclusion

Our results demonstrated that exosomes from serum are critical for maintaining the normal function of BMECs.

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Abbreviations

BMECs:

Bovine mammary epithelial cells

HS:

Heat stress

Ctr:

Contrast

qRT-PCR:

Quantitative real-time PCR

PBS:

Phosphate buffer solution

SDS:

Sodium dodecyl sulfate

TBST:

Tris-buffered saline containing 0.1% Tween 20

Exo:

Exosomes

ELF5:

E74-like Factor 5

CSN2:

β-Casein

STAT5:

Signal transducer and activator of transcription 5

SREBP1:

Sterol regulatory element binding protein 1

ADD1:

Adipocyte determination and differentiation factor-1

TG:

Triacylglycerol

AMPK:

Adenosine 5′-Monophosphate (AMP)-activated protein kinase

PI3K:

Phosphoinositide3-kinase

AKT:

Threonine-protein kinase

mTOR:

Mammalian target of rapamycin

ROS:

Reactive oxygen species

Mn-SOD:

Manganese superoxide dismutase

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Number 32002169), the Natural Science Foundation of Jiangsu Province (Grant Number BK20190254) and the Jiangsu Agricultural Science and Technology Innovation Fund (Grant Number CX (19)2037).

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Authors and Affiliations

  1. Institute of Animal Science/Key Laboratory for Crop and Animal Integrated Farming of Ministry of Agriculture and Rural Affairs, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China

    Yue Wang, Hui-Li Wang, Ji-Feng Zhong & Kun-Lin Chen

  2. Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Hangzhou, 311300, China

    Xing Duan

  3. Youyuan Research Institute of Dairy Industry Co., Ltd, Nanjing, 211100, China

    Hui-Li Wang, Zhi-Ping Lin, Ji-Feng Zhong & Kun-Lin Chen

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  1. Yue Wang
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Contributions

YW conceptualized the study and analyzed data. HLW, ZPL and JFZ carried out the molecular studies and sample collection. KLC and XD designed the research and drafted and revised the manuscript. All authors read and approved the final manuscript for publication.

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Correspondence to Kun-Lin Chen or Xing Duan.

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Wang, Y., Wang, HL., Lin, ZP. et al. Depletion of serum-derived exosomes aggravates heat stress-induced damage of bovine mammary epithelial cells. Mol Biol Rep 49, 9297–9305 (2022). https://doi.org/10.1007/s11033-022-07767-6

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