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The effects of autophagy in rat tracheal epithelial cells induced by silver nanoparticles

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Abstract

The massive use of silver nanoparticles (AgNPs) is potentially harmful to exposed humans. Although previous studies have found that AgNPs can induce cell autophagy, few studies have focused on the toxic pathways and mechanisms of autophagy induced by AgNPs in rat respiratory epithelial (RTE) cells. In this study, RTE cells were exposed to two kinds of AgNPs in vitro to ascertain the influence of mTOR-autophagy pathway-associated protein expression, including Beclin1, LC3B, Atg5, and Atg7. After exposure to different sizes and concentrations of AgNPs for 12 h, the uptake of silver in RTE cells reached 0.45 μg/L to 1.11 μg/L, indicating that AgNPs can enter RTE cells, leading to toxic effects. Our study found that this toxic effect was related to autophagy caused by ROS accumulation that was mediated by the mTOR pathway. With increasing AgNP exposure concentrations, the expression of p-mTOR was significantly downregulated, and expression of the autophagy-related proteins Beclin1, LC3B, Atg5, and Atg7 was significantly increased in RTE cells in all exposed groups. At a concentration of 1000 μg/L, the expression of LC3BII/LC3BI in all exposed groups was 24.49 times and 12.71 times that of the control, and the expression of Atg7 in all exposed groups was 23.21 times and 13.21 times that of the control. The upregulation of autophagy-related proteins in the AgNP-10 nm exposure groups was greater than that of the AgNP-100 nm exposure group. In summary, the mTOR pathway mediates AgNP-induced autophagy in RTE cells, which leads to damage to the respiratory system barrier and human health risks. This study can facilitate the development of prevention and intervention policies against adverse consequences induced by AgNPs.

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Data availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

RTE cell:

Rat tracheal epithelial cell

AgNPs:

Silver nanoparticles

FBS:

Fetal bovine serum

PBS:

Phosphate balanced solution

DMEM:

Dulbecco’s modified Eagle’s medium

DCFH-DA:

2,7-Dichlorodihydrofluorescein diacetate

CCK-8:

Cell counting kit-8

ICP-OES:

Inductively coupled plasma emission spectrometer

HRP:

Horseradish peroxidase

IgG:

Immunoglobulin G

MDA:

Malondialdehyde

MT:

Metallothionein

mTOR:

Mammalian target of rapamycin

T-mTOR:

Total mTOR

p-mTOR:

Phosphorylation of mTOR

LC3B:

Microtubule-associated protein light chain 3 B

Atg5:

Autophagy-related gene 5

Atg7:

Autophagy-related gene 7

mL:

Milliliter

rpm:

Revolutions per minute

min:

Minute

s:

Second

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Funding

This work was financially supported by the Natural Science Foundation of Zhejiang Province of China (No. LY15B070014) and Hangzhou Science and Technology Program (No. 20150533B02).

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Authors and Affiliations

  1. School of Life and Environmental Sciences, Hangzhou Normal University, Yuhangtang Road 2318#, Hangzhou, 311121, Zhejiang Province, China

    Hangjun Zhang, Feifei Chen, Yan Li, Xiaodong Shan, Lu Yin, Xiaojing Hao & Yuchi Zhong

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  1. Hangjun Zhang
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  2. Feifei Chen
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  3. Yan Li
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  4. Xiaodong Shan
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  5. Lu Yin
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  6. Xiaojing Hao
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  7. Yuchi Zhong
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Contributions

Hangjun Zhang, Yuchi Zhong, and Feifei Chen contributed to the conception of the study; Feifei Chen, Xiaodong Shan, Lu Yin, and Xiaojing Hao performed the experiment; Feifei Chen and Yan Li contributed significantly to analysis and manuscript preparation; Hangjun Zhang, Yuchi Zhong, Feifei Chen, and Yan Li performed the data analyses and wrote the manuscript; Hangjun Zhang and Yuchi Zhong helped perform the analysis with constructive discussions. All authors read and approved the final manuscript.

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Correspondence to Yuchi Zhong.

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Zhang, H., Chen, F., Li, Y. et al. The effects of autophagy in rat tracheal epithelial cells induced by silver nanoparticles. Environ Sci Pollut Res 28, 27565–27576 (2021). https://doi.org/10.1007/s11356-020-12259-w

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