Influence of Nitrofurantoin on Toll-Like Receptor Pathway in Human Trophoblastic Cells (Bewo) Infected with Toxoplasma Gondii

Abstract Toxoplasmosis is a common parasitic disease caused by the obligatory intracellular Toxoplasma gondii . To investigate the effect of nitrofurantoin (NF) on the signaling pathway of TLR (Toll-Like Receptor) in BeWo, a human trophoblastic cell line, infected with T. gondii . Primarily, to determine the concentration of NF, giemsa staining method, trypan blue staining method and MTS (3-(4,5-dimetiltiyazol-2-yl)-5-(3-karboksimetoksifenil)-2-(4-sülfofenil)-2H-tetrazolyum) test were used on negative-positive controls and experimental groups. After MTS measurement, IC50 (half maximal inhibitory concentration) and EC50 (half maximal effective concentration) values and the appropriate concentration as 1.25 µM were calculated using GraphPad Prism 8.0. In order to determine the effect of NF on the TLR pathway, samples were collected at three different times: 24, 48 and 72 hours. After RNA isolation, cDNA was synthesized and Real-Time PCR (RT-qPCR) analyzes were performed. Proportional changes in gene expression were calculated using reference genes. According to this, the most effective time period of the drug was found to be 48 hour period based on the decrease in expression levels in some apoptosis-related genes and, TLR6 (Toll-like receptor 6) and TLR8 (Toll-like receptor 8). There were significant differences after 48 (p = 0.015) and 72 (p = 0.049) hours in the parasitic load at experimental groups. As a result, it was shown that NF inhibited T. gondii infection by affecting genes in the TLR pathway. Furthermore, to know that these knowledges has shown the possibility that new preparations with less side effects can be developed as an alternative method in the treatment of the disease.