In Vitro Studies on Induction of Ferroptosis Upon Low‐level Exposure of Bisphenol A on Rat Erythrocytes

ABSTRACT Bisphenol A (BPA), is a xenoestrogen used in plastic manufacturing companies. Some countries are limiting the use of BPA. The widespread use of BPA has resulted in its accumulation in the natural environment. This compound is extensively utilised in various industrial products and is linked to harmful effects on both wildlife and human health. In recent years, extensive research has focused on ferroptosis—an iron‐dependent form of programmed cell death characterised by lipid peroxidation. Also, several studies highlight the involvement of ferroptosis in mediating the toxic effects of environmental pollutants. This study aimed to deepen our understanding of BPA‐induced ferroptosis by investing its impact on the antioxidant defence system, the structural integrity (as indicated by the rate of haemolysis), and the functionality of red blood cells. The research focused on evaluating the toxic effects of BPA through biochemical and haematological analyses conducted on RBCs of rats. To evaluate the involvement of ferroptotic cell death in BPA‐induced toxicity, ferroptosis inducer RSL‐3 and ferroptosis inhibitor Ferrostatin were used in the study either alone or in co‐administration with highest concentration of BPA. The results showed that compared to the vehicle treated control group, after acute exposure of RBCs to BPA showed a notable decrease in Hb (haemoglobin) content and number of red blood cells. The ferroptotic markers viz. lipid peroxidation, GPx4 levels and ACSL4 activity were observed across all treatment groups. The results showed the possible involvement of ferroptosis on all these markers which were confirmed in the presence of ferroptosis inducer and inhibitor. Also, oxidative and nitrosative stress levels increased in a concentration‐dependent manner of BPA. Furthermore, the activity of two important antioxidant enzymes; superoxide dismutase, and catalase lowered notably after BPA treatment. Results also showed significant morphological alterations, and increased membrane damage of RBC upon of BPA treatment. The study concludes that BPA exhibits oxidative stress, which may be one of the mechanisms causing RBC toxicity. The present study indicated that ferroptosis plays a key role in the progression of BPA‐induced RBC toxicity; thus it can become a novel target in treatment of haemolysis.