The effect of resveratrol on oxidative stress and Klotho expression in iopromide-induced human renal tubular epithelial HK-2 cells

ZHANG Bin; ZHONG Xiao-hua; LIAO Li-zhen

doi:10.3969/j.issn.1001-9448.2018.04.004

Abstract：Objective To investigate the effect of resveratrol on oxidative stress and Klotho expression in iopromide-induced human renal tubular epithelial HK-2 cells. Methods Different doses of iopromide ( 137. 5 mgI /mg, 275. 0 mgI /mg and 550. 0 mgI /mg) were added in human renal tubular epithelial HK-2 cells for 12 h to mimic contrast-induced nephropathy in vitro. MTT assay was used to measure the cell viability. Kit assay was used to measure lactate dehydrogenase enzyme ( LDH) vitality and flow cytometry was used to measure percentage of apoptosis. Different doses( 1 μmol /L, 10 μmol /L and 100 μmol /L) of resveratrol or N-acetylcysteine ( NAC) was added, and the cell viability, LDH vitality and percentage of apoptosis were detected. CM-H2DCFDA assay was used to measure cell fluorescence intensity of reactive oxygen species ( ROS) . Western bolt assay was used to measure the expression of Klotho protein. Results Iopromide treatment for 12 h in human renal tubular epithelial HK-2 cells caused cell viability decrease, and elevated LDH vitality and apoptosis in a dose-dependent manner. When different doses of resveratrol was added, the above cell changes were inhibited in a dose-dependent manner. The effect of resveratrol was similar to the NAC. In addition, iopromide treatment caused ROS fluorescence intensity upregulation, which was inhibited by both resveratrol and NAC. The Klotho protein expression was reduced in the iopromide group when compared with the control group, which was inhibited by resveratrol but not NAC. Conclusion Resveratrol alleviates iopromide-induced toxic effects on HK-2 cells as the same as NAC. It may be related to its effects on upregulation of Klotho protein and downregulation of oxidative stress in HK-2 cells.