Detection of ROS by flow cytometry.

RCECs were cultured in normal osmotic pressure(A), normal osmotic pressure added with NAC (B) or DPI (C), and 90 mM NaCl high osmotic pressure (HOP) group (D), 90 mM NaCl HOP added with NAC (E) or DPI (F). Intracellular ROS levels in RCECs were detected using DCFH-DA. Intracellular ROS levels increased in the HOP group. ROS levels decreases in the HOP added with NAC and DPI group. Statistics (G) showed that the fluorescence intensity in the 90 mM NaCl HOP group was significantly higher than that in the normal osmotic pressure group (P<0.01), and the fluorescence intensity in the HOP group added with ROS inhibitor NAC and DPI decreased to some extent. The decrease was statistically significant in the HOP group added with DPI(P<0.01). ** P<0.01 vs. A, ## P<0.01 vs. D.