HERP depletion inhibits zearalenone-induced apoptosis through autophagy activation in mouse ovarian granulosa cells. (February 2019)

• Record Type: Journal Article
• Authors: Chen, Fenglei; Wen, Xin; Lin, Pengfei; Chen, Huatao; Wang, Aihua; Jin, Yaping
• Is Part Of: Toxicology letters. Volume 301(2019)
• Page Start: 1
• Page End: 10
• Publication Date: 2019-02
• Subjects: 4-PBA 4-phenylbutyric acid -- 7-AAD 7-amino-actinomycin D 7 -- ANOVA one-way analysis of variance -- ATF activating transcription factor -- ATG autophagy-related proteins -- BAX BCL-2 associated X protein -- BCL-2 B-cell lymphoma-2 -- CHOP CCAAT/enhancer binding protein homologous protein -- CQ chloroquine -- DMEM/F12 Dulbecco's modified Eagle's medium: nutrient mixture F-12 -- DMSO dimethyl sulfoxide -- ECL enhanced chemiluminescence -- ELISA enzyme linked immunosorbent assay -- ER endoplasmic reticulum -- ERAD ER-associated protein degradation -- ERK1/2 extracellular signal-regulated kinase ½ -- FBS fetal bovine serum -- Fisher LSD Fisher's least significant difference test -- GRP78 glucose-regulated protein 78 -- HERP homocysteine-responsive ER-resident protein -- IRE1α inositol-requiring enzyme 1α -- LC3 microtubule-associated protein light chain 3 -- MTT 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide -- mTOR mammalian target of rapamycin -- PERK protein kinase (PKR)-like ER kinase -- pERK1/2 phosphorylation of ERK1/2 -- pS6K1 phosphorylation of S6K1 -- PVDF polyvinylidene difluoride -- S6K1 p70 ribosomal protein S6 kinase -- shHERP HERP shRNA -- shNC non-silencing negative control -- shRNAs short hairpin interfering RNAs -- SPSS Statistical Package for the Social Science -- UPR unfolded protein response -- ZEA zearalenone; HERP -- Autophagy -- Zearalenone -- Apoptosis -- Granulosa cells; Toxicology -- Periodicals; 363.179
• DOI: 10.1016/j.toxlet.2018.10.026
• Languages: English
• ISSNs: 0378-4274

Abstract

Highlights: ZEA decreases cell viability and increases cell apoptosis in ovarian granulosa cells in a dose-dependent manner. Autophagy and ER stress cooperate in cell apoptosis in ovarian granulosa cells that is induced by ZEA. HERP depletion inhibits ZEA-induced cell apoptosis in ovarian granulosa cells through autophagy activation and apoptosis inhibition. Abstract: HERP is an endoplasmic reticulum (ER) membrane protein and is strongly induced by stress conditions. A recent study has indicated that HERP cooperates in apoptosis during zearalenone (ZEA) treatment. However, regulatory mechanisms and the role of HERP in ZEA-induced apoptosis remain elusive in ovarian granulosa cells. In this study, MTT and flow cytometry assays demonstrated that ZEA gradually decreased cell viability and increased apoptosis in granulosa cells in a dose-dependent manner. Western blot analysis showed that ZEA significantly activated autophagy by upregulating LC3-II. Chloroquine (CQ) significantly increased LC3-II and induced granulosa cell apoptosis. Moreover, Western blot analysis showed that ZEA inhibited the mTOR and ERK1/2 signaling pathways. Furthermore, we found that ZEA activated ER stress by upregulating the ER stress-related proteins GRP78, HERP and CHOP. 4-PBA significantly decreased GRP78, HERP, CHOP and LC3-II. In addition, knockdown of HERP (shHERP) significantly protected ovarian granulosa cells from apoptosis induced by ZEA. We found that HERP depletion activated autophagy and ERK1/2 signaling pathways, while it inhibited the mTOR and caspase-dependent mitochondrial signaling pathways. In summary, autophagy and ER stress cooperated in apoptosis induced by ZEA; HERP depletion inhibits ZEA-induced apoptosis of ovarian granulosa cells through autophagy activation and apoptotic pathway inhibition.