WJ-39, an Aldose Reductase Inhibitor, Ameliorates Renal Lesions in Diabetic Nephropathy by Activating Nrf2 Signaling

Figure 4

WJ-39 alleviated oxidative stress in rat mesangial cells (RMCs) cultured under high glucose (HG) conditions. (a) Cell viability was detected by the MTT assay after cells were exposed to 30 mM HG for 24, 48, and 72 h. (b) Cell viability was detected by the CCK-8 assay after cells were exposed to 5.6 mM normal glucose (NG) for 48 h. The results are expressed as the percentage of surviving cells. (c, e) Reactive oxygen species (ROS) levels in cells were detected by using a DCFH-DA probe, and the data are expressed as fluorescence intensity. (d) RMCs were treated with different concentrations of Nrf2 siRNA (30, 50, and 100 nM), and the protein levels of Nrf2 were detected by western blotting and quantified. (f) Protein levels of thioredoxin (TRX), heme oxygenase-1 (HO-1), and NAD(P)H dehydrogenase [quinone] 1 (NQO1) were detected by western blotting and quantified. Data are represented as the (SEM), . ^# and ^## vs. the NG group; and vs. the HG group; ^& vs. the WJ-39 (10 μM)+HG+control siRNA group. MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; CCK-8: Cell Counting kit-8; DCFH-DA: dichloro-dihydro-fluorescein diacetate.

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