Diphlorethohydroxycarmalol Attenuates Methylglyoxal-Induced Oxidative Stress and Advanced Glycation End Product Formation in Human Kidney Cells

Figure 5

DPHC did not affect antioxidant, detoxifying, ROS, AGEs, or protein carbonyl content in Nrf2-knockdown HEK cells. HEK cells were transfected with Nrf2 siRNA for 36 h. The transfected cells were incubated with 40 μM DPHC for 24 h. (a) qRT-PCR and Western blot were performed for nuclear factor (erythroid-derived 2)-like 2 (Nrf2) mRNA and protein expression. (b) qRT-PCR was performed for superoxide dismutase (SOD) 1 and 2, catalase (CAT), heme oxygenase-1 (HO-1), NAD(P)H quinone dehydrogenase (NQO)1, glutamylcysteine synthetase (GCL)c, and GCLm. (c–g) HEK cells were transfected with Nrf2 siRNA for 36 h. The transfected cells were incubated with or without 40 μM DPHC for 1 h and then further incubated with or without 1 mM MGO for 24 h. (c) ROS production was measured by flow cytometry. (d) AGE content was measured. (e) Protein carbonyl content was measured. (f) Nrf2 mRNA expression was measured by qRT-PCR. (g) Glo-1 (glyoxalase 1) mRNA expression was measured by qRT-PCR. Experiments were performed in triplicate. , , and . n.s.: no significance.

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