Oxidative Medicine and Cellular Longevity / 2018 / Article / Fig 2

Research Article

Nicotine and Cotinine Inhibit Catalase and Glutathione Reductase Activity Contributing to the Impaired Osteogenesis of SCP-1 Cells Exposed to Cigarette Smoke

Figure 2

CSE induces oxidative stress with an increase in O2 and GSH reduction in SCP-1. SCP-1 cells were exposed to 5% CSE, and intracellular ROS and GSH levels were measured with different fluorescent probes: (a) DCFH-DA assay was used to detect O2, H2O2, HO, and ONOO. To trap different ROS, SCP-1 cells were coincubated with 25 μM α-tocopherol (O2 i), 10 mM sodium-pyruvate (H2O2i), 250 mM DMSO (HOi), or 100 μM uric acid (ONOO i); (b) DHE assay was used to detect O2, and (c) Ellman assay was used to detect total GSH levels. Results were normalized to control SCP-1 cells (Crl). 0.01% H2O2 was used as positive control (a) or negative control (b) of the assays. Each experiment was conducted at least four times independently with triplicate. The statistical significance was determined by the Kruskal-Wallis H test followed by Dunn’s posttest. Data are represented as the mean ± SEM, and the significance was represented as , , and vs the control group.

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