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Problem The intracellular antioxidant system, based on glutathione (GSH), plays a key role in endometrial detoxification reactions and has been proposed to be involved in the pathogenesis endometriosis. This study was designed to evaluate whether estradiol (E2) and proinflammatory cytokines have any effects on expression of glutathione in endometrial stromal cells (ESCs). Method of study Glutathione levels were measured utilizing high-performance liquid chromatography following in vitro culture and treatment of ESCs with estradiol, tumor necrosis factor-alpha (TNF-α) and interleukin 1-beta (IL-1β). Results The GSH level in E2 (10−8 m) treatment group was significantly higher than in the control group at 48 h ( P < 0.05). In vitro treatment of ESCs with TNF-α 10 ng/mL as well as E2 (10−8 m) plus TNF-α 10 ng/mL for 48 hr also led to a significant increase in GSH level ( P < 0.05; P < 0.05, respectively). Both IL-1β 10 ng/mL and E2 (10−8 m) plus IL-1β 10 ng/mL for 48 hr increased GSH level significantly ( P < 0.05; P < 0.05, respectively) as well. Conclusions These findings might suggest that increased production of estradiol and proinflammatory cytokines in the peritoneal cavity possibly leads to the establishment of endometriosis through increased level of GSH. [ABSTRACT FROM AUTHOR]