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Butylated hydroxytoluene induces dysregulation of calcium homeostasis and endoplasmic reticulum stress resulting in mouse Leydig cell death

2019; Elsevier BV; Volume: 256; Linguagem: Inglês

10.1016/j.envpol.2019.113421

1873-6424

Jiyeon Ham, Whasun Lim, Kwang‐Youn Whang, Gwonhwa Song,

Carcinogens and Genotoxicity Assessment

Butylated hydroxytoluene (BHT) is a synthetic phenolic antioxidant that has been used as an additive for fat- or oil-containing foods. The exposure index value increases with extended usage of the chemical. Further, estimated total amount of BHT could exceed standard regulation, considering dietary intake or another exposure. Although BHT may induce side effects in reproductive systems, adequate research had not yet been performed to confirm them. In this study, we investigated the effects of BHT on mouse Leydig cells (TM3), which are components of testis. Our results indicated that BHT suppressed cellular proliferation and induced cell cycle arrest in TM3 cells. Moreover, BHT hampered cytosolic and mitochondrial calcium homeostasis in TM3 cells. Furthermore, BHT treatment led to endoplasmic reticulum (ER) stress and DNA fragmentation , simultaneously stimulating intrinsic apoptosis signal transduction . To elucidate the mode of action of BHT on Leydig cells, we performed western blot analysis and confirmed the activation of the PI3K/AKT and MAPK pathways. Collectively, our results demonstrated that BHT has toxic effects on mouse Leydig cells via induction of calcium dysregulation and ER-mitochondria dysfunction. Hypothetical mechanism of BHT in TM3 cells. Possible mode of action for BHT in TM3 including the MAPK, PI3K, and ER stress signaling pathways. Abbreviations: N, nuclei; Mito, mitochondria; ER, endoplasmic reticulum. [Ca 2+ ]i and [Ca 2+ ]m refer to cytosolic and mitochondrial calcium ion concentrations, respectively. • BHT induces cell cycle arrest and apoptosis of mouse Leydig cells. • BHT interferes cytosolic and mitochondrial calcium homeostasis in mouse Leydig cells. • BHT dysregulates genes related to steroidogenesis in mouse Leydig cells. • BHT leads to ER stress and DNA fragmentation of mouse Leydig cells. Butylated hydroxytoluene induces mouse Leydig cell death by suppressing the cell cycle and stimulating ER stress and mitochondrial apoptosis-related proteins.