Bisphenol a Disrupts Steroidogenesis and Induces Apoptosis in Human Granulosa Cells Cultured In Vitro

Bisphenol A (BPA) is a common synthetic chemical compound classified as an endocrine disruptor. It affects multiple physiological systems in the body, including the female reproductive system, particularly granulosa cells (GCs) in the ovaries, where steroidogenesis occurs. This study investigated the impact of various BPA concentrations (environmentally relevant concentrations of 0.001 µM and 0.1 µM and toxicological concentration of 100 µM) and exposure times (24 and 72 h) on cell viability and counts and in vitro production of estradiol and progesterone in human GCs collected from waste follicular fluid of IVF patients. Gene expression analysis of 182 genes associated with steroidogenesis and apoptosis was performed in GCs using PCR arrays, followed by protein expression analysis by Western blot. Our results demonstrate that after longer BPA exposure (72 h), a higher concentration of BPA (100 µM) negatively affects the cellular viability and counts and significantly alters steroid hormone biosynthesis in vitro, leading to reduced concentrations of estradiol and progesterone in the culture medium. We found that all BPA concentrations altered the expression of different steroidogenesis- and apoptosis-related genes in GCs. At 0.001 μM, BPA exposure decreased the expression of <i>TRIM25</i>, <i>UGT2B15</i>, <i>CASP3</i>, and <i>RPS6KA3</i> genes and increased the expression of <i>NR6A1</i> and <i>PPID</i> genes. At 0.1 μM, BPA increased the expression of <i>AR</i>, <i>HSD3B1</i>, <i>BID</i>, <i>IKBKG</i>, and <i>PPID</i> genes while reducing the expression of <i>TRIM25</i> and <i>CASP3</i> genes. At the highest concentration of 100 μM, BPA upregulated the expression of <i>AR</i>, <i>GPER30</i>, <i>BID</i>, <i>IKBKG</i>, and <i>PPID</i> genes and downregulated the expression of <i>FOXO1</i> and <i>UGT2B15</i> genes. These results highlight BPA's concentration-specific effects on steroidogenesis and apoptosis and show its potential to compromise GC function, with possible negative implications for female fertility and ovarian health, even at environmentally relevant concentrations.