Alternative targets of pesticide toxicity: effect of chlorpyrifos in TLR expression in human trophoblasts

GOMEZ DIEGO SEBASTIAN; GUIÑAZU ALANIZ, NATALIA LORENA

Mar del Plata

Congreso; LXIV Reunión Anual de la Sociedad Argentina de Investigacion Clínica (SAIC) - Reunión Anual de Sociedades de Biociencias 2019; 2019

Sociedad Argentina de Investigacion Clinica

Organophosphate (OP) pesticides family have been widely used and even today are still among the most commonly used type of pesticide. It has been widely studied their cholinergic mechanism as well as their impact in the placenta. Besides, it has been proposed OPs may alter innate immune system as an alternative toxic mechanism. The human placenta is a highly specialized organ composed of both maternal and fetal cells that performs vital during gestation, in which tolerance and immune response are a key factor in outcome. In this sense, Toll-Like Receptors (TLRs) are an essential component of the innate immune response. It has been identified TLRs in trophoblast (TLR-2, TLR-4) suggesting that the placenta can act as part of innate immune response. The objective of this work was to evaluate the impact of the OP clorpyriphos (CP) in the expression of toll like receptors in first trimester tophoblast.First trimester trophoblast cell line HTR-8/SVneo was exposed to CP (0-100 µM) for 24 h. TLRs transcript expression levels (TLR-2 and TLR-4) were studied. Quantitative Polymerase Chain Reaction assays were performed and TLRs expression levels were normalized with 18 s as housekeeping gene. In addition, protein levels of TLR-4 after exposure were evaluated by western blot in 10% SDS-PAGE. β-actin was used as loading control. Statistical differences were analyzed by Dunnet´s pos hoc of three independent experiments.Preliminary results indicated that CP alters the expression of both TLR-2 and TLR-4.Higher CP concentration increased TLR-2 mRNA expression up to 8 folds compared to control values, whereas same CP concentration augmented TLR-4 transcript levels up to 3 folds compared to control values. TLR-4 protein levels did not show significant differences compared to control as a result of CP treatment.The increments in TLRs transcript are indicative of the impact of the OPs clorpyriphos in the innate immune response in first trimester trophoblast. Nevertheless, in order to understand this scenario is important to study the balance of cytokines as an indicator of inflammatory response in tophoblasts exposed to OP.