The aim of this study was to investigate the effects of nonylphenol (NP) exposure on the expression of cell receptors and secretory function in mouse Sertoli TM4 cells. There were no significant changes in mRNA expression of estrogen receptor (ER)-α and toll like receptor (TLR)-4 in the cells exposed to NP for 24h. However, the mRNA expression levels of ER-β, progesterone receptor (PR) and androgen receptor (AR) were down-regulated in NP groups. Furthermore, NP treatment evoked significant changes in protein expression levels of ER-β and follicle-stimulating hormone receptor (FSHR). There were significant changes in the mRNA expression levels of vinculin, N-cadherin and occludin, but not vimentin. Levels of inhibin B, androgen binding protein (ABP) and transferrin (Trf) were found to change significantly in NP challenged cells. Additionally, the decrease of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) mRNA expression and increase of cytokine levels were simultaneously found in NP stimulated TM4 cells. In conclusion, these findings have shown that NP exposure affected expression of cell receptors and may damage specific secretory function of Sertoli TM4 cells, which may be associated with the male-specific reproductive toxicity of NP.