Experimental Nephrology and Genetics: Research Article
Zhang X. · Wang Q. · Li F. · Li S. · Lin H. · Huo Y.
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Article / Publication Details
Abstract
Introduction: We here evaluated the efficacy of piceatannol (PIC) in high glucose (HG)-induced injury of renal tubular epithelial cells HK-2.
Methods: After the establishment of HG-induced cell injury model and the treatment with PIC at both high and low concentrations and/or Acetazolamide (ACZ, the inhibitor of carbonic anhydrase 2 (CA2)), MTT and flow cytometry assays were carried out to confirm the viability and apoptosis of HK-2 cells. The levels of oxidative stress markers lactate dehydrogenase (LDH), malondialdehyde (MDA), and reactive oxygen species (ROS), the ratio of glutathione/oxidized glutathione (GSH/GSSG), and the CA2 activity were determined. Both quantitative reverse-transcription polymerase chain reaction and Western blot were used to calculate the expressions of CA2 (the predicted target gene of PIC via intersecting the data from bioinformatic analyses), and AKT pathway- (Phosphatase and tensin homolog (PTEN), phosphorylated (p)-AKT, AKT) and apoptosis-related proteins (Bcl-2 and cleaved caspase-3).
Results: HG suppressed cell viability and the levels of GSH/GSSG ratio, CA2, pThr308-AKT/AKT, pSer473-AKT/AKT, and Bcl-2, while promoting cell apoptosis, the levels of LDH, MDA, and ROS and the expressions of PTEN and cleaved caspase-3. All effects of HG were reversed by PIC at a high concentration. CA2 was predicted and identified as the target of PIC. In HG-treated HK-2 cells, additionally, ACZ reversed the effects of PIC on the viability, apoptosis, and the levels of both oxidative stress markers and AKT pathway- and apoptosis-related factors.
Conclusion: PIC protects against HG-induced injury of HK-2 cells via regulating CA2.
S. Karger AG, Basel
Article / Publication Details
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