Bisfenol A’nın SH-SY5Y Nöroblastoma Hücrelerindeki Asetilkolinesteraz Gen Ekspresyonuna Olan Etkilerinin Nitrik Oksit Sentaz Aracılığıyla İncelenmesi

Abstract

In this study, SH-SY5Y human neuroblastoma cells treated with low doses of Bisphenol A (BPA). Bisphenol A is a commonly used endocrine disruptor. The cytotoxicity of BPA to SH-SY5Y cells was determined by MTT and LDH assays, and NO levels were determined by the Griess method. The acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) gene expression levels of the cells were determined by RT-PCR and enzyme activities by the Ellman method. % Populations of cells were determined by Tali cytometer and TNFα and caspase-8 levels were measured with Elisa kits. It was determined that % viability of cells gave a non-monotonic dose response and a correlation between cytotoxicity results and NO levels. The IC50 values for AChE and BuChE enzymes were found to be 2.2 mM and 1.98 mM, respectively. The AChE gene expression level with 1 pM and 1 nM BPA at 48 hours was changed to 0.61 and 1.03 respectively and BuChE gene expression level was changed to 2.18 and 0.73 in comparison with the control. 1 pM BPA and 1 nM BPA decreased TNF-α levels to 0.073 and 0.084 respectively, at 48 hours. At 24 hours, it was determined that only 1 pM BPA treatment decreased TNF-α levels to 0.302. Caspase-8 levels were increased to 168.4%, 297.3% and 287%, 160.2% at 24 and 48 hours with 1 pM and 1 nM BPA, respectively. The apoptotic % cell population with 1 pM BPA and 1 nM BPA increased sequentially at 6 hours (7.3%, 7.7%) and decreased at 48 hours (1.3%, 1%) while the necrotic cell population decreased in 4. and 6. hours (20.3%, 18.3% -19.3%, 21.3%) and increased in 48 hours (23.7%, 27%). The results of this study demonstrate that cell switches from apoptosis to a necroptosis, a programmed death, at 48 hours by applying 1 pM and 1 nM BPA to SH-SY5Y cells.