Skip to main content
. 2017 Sep 26;7(11):1672–1685. doi: 10.1002/2211-5463.12284

Figure 4.

Figure 4

Advanced glycation end products induce intracellular reactive oxygen species (ROS) level and NADPH oxidase activity. (A) Images of control and AGE‐treated NRCMS stained with ROS‐sensitive dye DCF‐DA. (B) Quantification of fluorescence intensity indicated a significantly higher ROS level in NRCM treated with AGE (1 mg/mL, 24 h) (P < 0.001, n = 38–48 cells). (C) NADPH oxidase activity measurement in AGE‐treated and control NRCMs showed that AGE‐treated NRCM exhibited a significant increase (P < 0.01) in NADPH oxidase activity compared to the control cells (n = 7–9). (D) Addition of NADPH oxidase inhibitor diphenyleneiodonium (DPI, 25 μm) inhibited the difference between control and AGE‐treated NRCM (n = 5). (E) Examples of calcium traces from control and AGE‐treated NRCMs (1 mg/mL, 24 h) in the presence of 1 μm DPI. (F) Quantification of calcium amplitude showed no difference between control and AGE + DPI‐treated NRCM, indicating that DPI treatment suppressed the effect of AGE. (G) NRCM viability following treatment with AGE (1 mg/mL, 24 h) was assessed using MTT assay. There was no difference in the MTT colorimetric signal intensity between control and AGE‐treated cells (n = 12 in each group), suggesting that there was no difference in cell viability between these groups. (H) Consistently, analysis of caspase 3/7 activity showed that there is no difference in apoptosis level between AGE‐treated and control cells (n = 12 in each group).