(1056) The anti-inflammatory and antioxidant effects of exogenous oxytocin on human placental cells

Inflammatory response:  JAR and JEG-3 cells (ATCC) were plated at 2x10⁵ cells per mL: JAR cells were plated in RPMI 10% Fetal bovine serum (FBS) PSQ and JEG-3 cells were plated in DMEM 10%FBS PSQ. All assays were performed in triplicate and repeated three times. At confluence, cells were treated with vehicle (HBSS), OXT (,125500nM) or N-acetyl cysteine (NAC, 5mM). After 24 hours, the cells were then washed, collected, resuspended in 2 mL HBSS and labeled with 20 µM DCF-DA. After 24 hours, cells were treated with TNF (20ng/ml) to induce a pro-inflammatory response. After 24 hr., culture supernatants were analyzed for IL-6 by ELISA.

Oxidative stress: The cells were set up as described above and were then treated with vehicle (HBSS) or OXT (125-500nM). The plates were read in a VICTOR3™ plate reader after 10 min (basal reading). Hydrogen peroxide (H₂O₂) was then added to induce oxidative stress and the plates were re-read 1 hr. later.  

Cell viability: JAR and JEG-3 cells were plated in media as described above.  The cells were then treated with vehicle (HBSS) or OXT (125-500nM) and cytotoxicity was measured using the neutral red assay 48 hr. later.

Results: OXT does not induce cytotoxic effects on JAR or JEG-3 cells. OXT significantly reduced TNF induced IL-6 production by JAR (p=0.006) and JEG-3 (p=0.002) cells (Figure 1A-B). Similar to NAC (a well-known antioxidant), OXT exerts dose-dependent antioxidant effects by JAR and JEG-3 cells under basal and following H₂O₂ stimulation (p < 0.001; Figure 1C-F). The anti-inflammatory and antioxidant effects of OXT are not accompanied by cytotoxicity.

Conclusion: Exogenous OXT has strong anti-inflammatory and antioxidant effects on human placental cell lines.