RESUMO
This study was performed to evaluate the effect of monobutyl phthalate (MBP) on GPR30-activated pathways in Sertoli cells. Additionally, we tested if GIM-1 (Panax ginseng metabolite) modulates MBP action. Human Sertoli cells (HSeC lineage) were exposed to MBP and/or GIM-1 for 30 min, 1, 12, and 48 h. Four experimental treatments were performed: control (DEMEM/F12 medium), MBP, GIM-1, and MBP + GIM-1. The results indicate that MBP activates GPR30, PKA, Src, EGFR, and the ERK1/2 proteins, while GIM-1 inhibits PKA, Src, ERK1/2, and the AKT pathway. MBP also enhances Cofilin expression, decreasing F-actin intensity on the cell surface in a short time. The combined exposure demonstrated a functional antagonism between compounds. Collectively, these data show that MBP activates GPR30 in Sertoli cells, and GIM-1 modulates this response, playing a protective role in Sertoli cells exposed to MBP.
Assuntos
Citoproteção/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Panax , Ácidos Ftálicos/toxicidade , Receptores de Estrogênio/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Células de Sertoli/efeitos dos fármacos , Linhagem Celular , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Humanos , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Células de Sertoli/metabolismo , Quinases da Família src/metabolismoRESUMO
This study evaluated oxidative stress markers in Human Sertoli cells cultivated on Geltrex® and exposed to Monobutyl Phthalate (MBP), and the potential cytoprotective role of GIM-1 on the antioxidant response. Exposure was performed at 30 min, 1, 12 and 48 h into 4 groups: control, MBP (10µM), GIM-1 (0,05µM) and MBP + GIM-1. Morphology was evaluated. Antioxidant enzymes were analyzed by colorimetric method; NRF-2, SIRT-1, 8- OHdG and Cleaved Caspase-3 by Western Blot. Larger spaces between cells were shown in MBP treatment; GIM-1 was similar to Control and MBP + GIM-1 showed an intermediate aspect. MBP reduced enzymatic activity of all enzymes and NRF-2 expression, increasing cleaved Caspase-3 expression; while GIM-1 increased antioxidants markers alone and attenuated MPB effects in MBP + GIM-1. MBP induced deleterious effects on Sertoli cells, increasing the oxidative stress, apoptosis and modifying their distribution in culture; however, GIM-1 acted as an important cytoprotective agent reversing our attenuating MBP effects.