(163) Tolylfluanid Disrupts Implantation of Blastocysts by Inducing Cell Death Through the Generation of Reactive Oxygen Species in Porcine Trophectoderm and Uterine Luminal Epithelial Cells

Abstract Authors: Tolylfluanid Disrupts Implantation of Blastocysts by Inducing Cell Death Through the Generation of Reactive Oxygen Species in Porcine Trophectoderm and Uterine Luminal Epithelial Cells

D. Song¹, T. Hong¹, S.M. Bae², G. Song³, F.W. Bazer⁴, W. Lim^1,2

1. Department of Biological Sciences, Sungkyunkwan University, Suwon, Republic of Korea

2. Department of MetaBioHealth, Sungkyunkwan University, Suwon, Republic of Korea

3. Department of Biotechnology, Korea University, Seoul, Republic of Korea

4. Department of Animal Science, Texas A&M University, College Station, TX, USA

Abstract Text: Tolylfluanid Disrupts Implantation of Blastocysts by Inducing Cell Death Through the Generation of Reactive Oxygen Species in Porcine Trophectoderm and Uterine Luminal Epithelial Cells

D. Song¹, T. Hong¹, S.M. Bae², G. Song³, F.W. Bazer⁴, W. Lim^1,2

1. Department of Biological Sciences, Sungkyunkwan University, Suwon, Republic of Korea

2. Department of MetaBioHealth, Sungkyunkwan University, Suwon, Republic of Korea

3. Department of Biotechnology, Korea University, Seoul, Republic of Korea

4. Department of Animal Science, Texas A&M University, College Station, TX, USA

Tolylfluanid, a pesticide and antifoulant used in agriculture and marine industries to prevent the attachment of fungi and marine organisms, has fungicidal effects on various crops. However, it is known to be a potential endocrine disruptor and may negatively impact placental development and reproductive health. This study investigated effects of tolylfluanid on implantation of blastocysts using porcine trophectoderm (pTr) and uterine luminal epithelial (pLE) cells as a model system. Tolylfluanid significantly reduced the survival rates of both pTr and pLE cells. Various flow cytometry analyses indicated that tolylfluanid increased the generation of reactive oxygen species (ROS), induced cell cycle arrest, and disrupted calcium homeostasis, leading to mitochondrial dysfunction. Moreover, the Mito Stress Assay confirmed that tolylfluanid significantly reduced the capacity for ATP production by mitochondria in both cell lines. Additionally, western blot analysis revealed that tolylfluanid altered the expression of proteins involved in the MAPK/PI3K pathway and apoptosis, thereby inducing cell death. The migratory ability of pTr and pLE cells was also diminished which adversely affected interactions between pTr and pLE cells. The generation of ROS induced by tolylfluanid was alleviated by addition of N-acetylcysteine, an ROS scavenger, to culture medium to improve cell viability and cell migration. In summary, these results indicate that tolylfluanid has the potential to interfere with the implantation of blastocysts by inducing cell death in both pTr and pLE cells.