Zearalenone Induces Apoptosis in Porcine Endometrial Stromal Cells through JNK Signaling Pathway Based on Endoplasmic Reticulum Stress

Toxins (Basel). 2022 Nov 3;14(11):758. doi: 10.3390/toxins14110758.

Abstract

Zearalenone (ZEA) is an estrogen-like mycotoxin characterized mainly by reproductive toxicity, to which pigs are particularly sensitive. The aim of this study was to investigate the molecular mechanism of ZEA-induced apoptosis in porcine endometrial stromal cells (ESCs) by activating the JNK signaling pathway through endoplasmic reticulum stress (ERS). In this study, ESCs were exposed to ZEA, with the ERS inhibitor sodium 4-Phenylbutyrate (4-PBA) as a reference. The results showed that ZEA could damage cell structures, induce endoplasmic reticulum swelling and fragmentation, and decreased the ratio of live cells to dead cells significantly. In addition, ZEA could increase reactive oxygen species and Ca2+ levels; upregulate the expression of GRP78, CHOP, PERK, ASK1 and JNK; activate JNK phosphorylation and its high expression in the nucleus; upregulate the expression Caspase 3 and Caspase 9; and increase the Bax/Bcl-2 ratio, resulting in increased apoptosis. After 3 h of 4-PBA-pretreatment, ZEA was added for mixed culture, which showed that the inhibition of ERS could reduce the cytotoxicity of ZEA toward ESCs. Compared with the ZEA group, ERS inhibition increased cell viability; downregulated the expression of GRP78, CHOP, PERK, ASK1 and JNK; and decreased the nuclear level of p-JNK. The Bax/Bcl-2 ratio and the expression of Caspase 3 and Caspase 9 were downregulated, significantly alleviating apoptosis. These results demonstrate that ZEA can alter the morphology of ESCs, destroy their ultrastructure, and activate the JNK signaling via the ERS pathway, leading to apoptosis.

Keywords: JNK signaling pathway; endoplasmic reticulum stress; porcine endometrial stromal cells; zearalenone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Caspase 3 / metabolism
  • Caspase 9 / metabolism
  • Endoplasmic Reticulum Stress*
  • MAP Kinase Signaling System
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Stromal Cells / metabolism
  • Swine
  • Zearalenone* / metabolism
  • bcl-2-Associated X Protein / metabolism

Substances

  • Zearalenone
  • 4-phenylbutylamine
  • Caspase 3
  • Caspase 9
  • bcl-2-Associated X Protein
  • Proto-Oncogene Proteins c-bcl-2

Grants and funding

This work was supported by the Project of Modern Agricultural Industry and Technology System of Anhui Province [grant number AHCYJSTX-05-03]; the Natural Science Foundation of Anhui Province [grant number 2208085MC81]; the College Students Innovation and Entrepreneurship Training Program of Anhui Province [grant number S202110364260]; and the College Students Innovation and Entrepreneurship Training Program of Anhui Agricultural University [grant number X202210364045]. The sponsors had no role in the study design; in the collection, analysis, and interpretation of the data; in the writing of the report; and in the decision to submit the article for publication.