Effects of oxidants and antioxidants on proliferation of endometrial stromal cells

Objective: To evaluate the effects of oxidative stress and antioxidants on proliferation of endometrial stromal cells. Design: In vitro study. Setting: Academic laboratory. Patient(s): Women, with and without endometriosis, of reproductive age. Intervention(s): Culture of endometrial stromal cells with antioxidants or with agents inducing oxidative stress. Main Outcome Measure(s): Proliferation of endometrial stromal cells as determined by thymidine incorporation assay and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay. Result(s): Antioxidants induced a dose-dependent inhibition of thymidine incorporation: vitamin E succinate was inhibitory at 10-100 muM (by 43%-95%), ebselen at 10-30 muM (by 29%-77%), and N-acetylcysteine at 10-30 mM (by 52%-85%). In contrast, modest oxidative stress induced by hypoxanthine/xanthine oxidase (I mM/3-30 muU/mL) stimulated proliferation by 40%-62%. H2O2 (1 muM) increased DNA synthesis by 56%. Comparable findings were obtained using MTT proliferation assay. Antioxidants inhibited proliferation: vitamin E succinate (100 muM) by 91%, ebselen (30 muM) by 81%, and N-acetylcysteine (30 mM) by 95%. Hypoxanthine/xanthine oxidase (1 mM/30 muU/mL) and H2O2 (1 muM) stimulated growth by 122% and 58%, respectively. Conclusion(s): Reactive oxygen species may modulate growth of endometrial stroma. Under pathologic conditions such as endometriosis, increased oxidative stress and depletion of antioxidants may contribute to excessive growth of endometrial stromal cells. (C) 2004 by American Society for Reproductive Medicine.