Figure 4.
Effects of cHSP60 on ROS-related enzymes and mitochondrial membrane potential in HCAECs. Cells were treated with cHSP60 or HI-cHSP60 (2 µg/mL) or superoxide generator LY83583 (10 µM) in the presence or absence of antioxidant SeMet (20 µM) or MnTBAP (2.5 µM) for 24 h. (A) The mRNA levels of ROS-related enzymes (NOX1, NOX4, p22phox, CAT, SOD1, GPX1, and GPX7) were determined by real-time PCR. (B) NADPH oxidase activity assay. NAPDH, Tiron (intra cellular scavenger of superoxide anion) and DPI (a flavoprotein inhibitor of NADPH oxidase) were included in the assay. (C) SOD activity assay. (D) CAT activity assay. (E) JC-1 staining and flow cytometry analysis for mitochondrial membrane potential. (F) ATP contents were determined by ATPLite kit. *P < 0.05, **P < 0.01, n = 3, t-test.