Figure 4. Effects of baicalin pretreatment following H₂O₂ stimulation on ROS production in bovine MEC.

BMECs were cultured for 24 h in medium without (control) or with different concentrations of H₂O₂ at increasing doses (0, 25, 50, 100, 200, 500, 1000 µM) in triplicate. Data shown as mean ± S.E. (A) BMECs were plated in 96 well plates at a density of 5,000/well. MTT (25 μl of five mg/mL) was added to check the cell viability assay (Optical density, OD). *p < 0.05 versus control (B) The ROS production was determined by a fluorimetric assay. Data are expressed in percentage of the control. *p < 0.05 versus control (C) and (D) BMECs were pretreated with different concentrations of baicalin (BC, 0–200 µg/mL). After 24 h, they were exposed to different concentrations of H₂O₂ (0, 25, 50, 100 µM) in triplicate (C) or to 100 µM in triplicate (D). The ROS production (C) was determined by a fluorimetric assay. Data are expressed in arbitrary unit (OD ×1000). *p < 0.05 versus control. Cell viability assay was also assessed (D). Results without Baicalin and with 100 µM H₂O₂ represent 100% viability. *p < 0.05 versus control.