Fig. 4

Enhanced ROS production is linked to hydrogen treatment in endometrial cancer cells. a Hydrogen treatment in AN3CA, HEC1A, Ishikawa cells induced decreased cell viability compared to CM cultured cells in a time-dependent manner at 48,72,96 and 120 h measured spectrophotometrically at 490 nm by MTT assay. b Upregulation of ROS as detected by the DCFH-DA method, AN3CA endometrial cancer cells, HEC1A cells, and AN3CA cells showed higher ROS levels upon treatment with hydrogen after 48 h, as recorded with mean FL1-A, and further decreased with NAC treatment. c Upregulation of ROS was observed in H-CM-treated HEC1A cells compared with the CM group. d Generation of mtROS was upregulated in H-CM-treated Ishikawa, HEC1A, and AN3CA cells compared with the CM group as measured with the MitoSOX™Red assay method and index ratios. e Ishikawa cells treated with MCC950 and NAC showed decreased expression of mtROS, while mtROS levels were upregulated in the LPS-primed group compared with the H-CM group. Data are expressed as mean ± S.D. NAC (5 mM) blocked the generation of ROS. The NLRP3 inhibitor MCC950 (10 nM) is known to potently inhibit NLRP3, and LPS (100 ng/mL) can stimulate NLRP3. Data are expressed as mean ± S.D. *, ** P < 0.05 compared with the control group. Each experiment was performed in triplicate (See Additional file 6)