The effect of pathogen-associated molecular patterns on the swine granulosa cells.

Recent studies have demonstrated the surprising ability of reproductive endocrine cells to express receptors of innate immunity useful to sense danger in order to avoid disruption of tissue homeostasis. Present research demonstrates the presence of pattern recognition receptors, i.e. toll like receptors (TLR) TLR2, TLR4 and TLR 5 and NOD like receptors (NLR) NOD1 and NOD2 in swine granulosa cells from ovarian follicles> 5 mm. Therefore, our second goal was to expose granulosa cells to different concentrations (1000, 100 and 10 ng/ml) of lipopolysaccharide (LPS) and N-Palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)- propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]- lysine (Pam3CSK4), two substances associated with pathogen molecular patterns. Their potential effects on the main functional parameters were monitored: proliferation (through the incorporation of Bromo-deoxy-Uridine), cell viability (by testing the metabolization of MTT salt), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test, and the non-enzymatic reducing power, by FRAP test). The data collected show a significant inhibition (p < 0.01) of cell proliferation after treatment with both LPS and Pam3CSK4, while cell viability has not been modified. As for steroidogenesis, treatment with both LPS and Pam3CSK4 significantly inhibited (p < 0.05) the production of 17β-estradiol and progesterone. LPS and Pam3CSK4 stimulated (p < 0.05) the production of superoxide anion and nitric oxide, while inhibited (p < 0.05) the antioxidant power. In conclusion, the study shows that the functionality of granulosa cells is compromised by the exposure to molecular profiles associated with pathogens; the knowledge gathered could lay the theoretical basis for the definition of therapeutic treatments related to diseases that can affect normal reproductive processes.