Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: Inhibition of thioredoxin 1 induces renal tubular cell oxidative injury. (A) Effects of Px-12 on cellular morphology. NRK-52E cells in 96-well plates were exposed to different concentrations of Px-12 (0, 5, 10, and 15 µM) for 24 h. Cell morphology was determined using an inverted microscope (×100). (B) Change of cellular viability induced by Px-12. NRK-52E cells in 96-well plates were insulted with different concentrations of Px-12 (0, 5, 10, and 15 µM) for 24 h. Then, cell viability was tested using a Cell Counting Kit-8 (CCK-8) assay. Data are expressed as the percentage of living cells vs. the untreated control (mean ± SD, n = 4). * p < 0.01 vs. the control. (C) Px-12 triggers apoptosis. NRK-52E cells in 96-well plates were exposed to different concentrations of Px-12 for 24 h. Then, cells were lyzed, and total protein was extracted. Caspase-3 and cleaved caspase-3 are detected via western blot analysis. Densitometric analysis of cleaved caspase-3 is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control). (D) Implications of Px-12 on reactive oxygen species (ROS)/O 2 − generation. NRK-52E cells in six-well plates were exposed to Px12 (10 µM) for 1 h, then incubated with a ROS/O 2 − probe for 30 min. Images have been acquired using an inverted fluorescence microscope. Mean fluorescence intensity is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control.).

Article Snippet: Intracellular ROS and O 2 − were determined using the ROS/O 2 − reagent kit (Enzo Life Sciences, New York, United States).

Techniques: Inhibition, Inverted Microscopy, Cell Counting, CCK-8 Assay, Western Blot, Incubation, Fluorescence, Microscopy

Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: Connexin 43 (Cx43) inhibition suppresses oxidative stress by regulating thioredoxin 1. (A) Effect of 18α-glycyrrhetinic acid (Ga) and sp600125 on reactive oxygens species (ROS)/O 2 − production. NRK-52E cells in a 96-well plate were pretreated with Ga and sp600125 separately for 1 h, challenged with Px-12 for another 1 h, and then incubated with a ROS/O 2 − probe for 30 min. Images were captured using an inverted fluorescence microscope (×100). Mean fluorescence intensity is shown at the bottom (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control). (B) Effects of Ga on thioredoxin 1. NRK-52E cells in six-well plates were pretreated with Ga for 1 h and then challenged with Px-12 for another 1 h. Then, cellular lysates were analyzed by western blotting for the expression of thioredoxin 1. Densitometric analysis of thioredoxin 1, as determined by ImageJ, is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the siRNA control group).

Article Snippet: Intracellular ROS and O 2 − were determined using the ROS/O 2 − reagent kit (Enzo Life Sciences, New York, United States).

Techniques: Inhibition, Incubation, Fluorescence, Microscopy, Western Blot, Expressing