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Protective Effect of Resveratrol on Mitochondria and Oxidative Stress induced by Cisplatin in HK‐2 Cells
Author(s) -
Valentovic Monica,
Murphy Rachel,
Lamyaithong Andre Benja,
Stafford Reagan,
Schnelle Anthony,
Ball John G
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.711.2
Subject(s) - cisplatin , viability assay , oxidative stress , chemistry , resveratrol , mtt assay , cytotoxicity , pharmacology , apoptosis , toxicity , biochemistry , medicine , in vitro , chemotherapy , organic chemistry
Cisplatin is used in treating many cancers including testicular, ovarian and cervical cancer. Impaired renal function is a major side effect of cisplatin and occurs in 33% of patients treated with cisplatin. Resveratrol (RES) is a polyphenolic compound that possesses anticancer activity. Part of the mechanism for reducing cisplatin toxicity by RES may be mediated by maintaining mitochondrial function and reducing oxidative stress. The purpose of this study was to evaluate the effect of cisplatin on mitochondrial function in a human proximal tubular kidney cell line (HK‐2). Specifically the levels of ATP and ADP were measured following exposure to cisplatin in the presence of RES or DMSO (vehicle). HK‐2 cells were pretreated for 1 h with 0–15 uM RES (vehicle control 10% DMSO). Renal cells were subsequently exposed to cisplatin at a final concentration of 0–30 uM for 24 or 48 h. Viability was assessed using MTT release and cell count with an n=6/treatment group and repeated 3 separate times. Cells pretreated with 10 uM RES were protected from cisplatin cytotoxicity at 24 and 48 h. Cisplatin caused a concentration dependent decline in MTT viability after 24 h exposure to 0, 7.5 and 15 uM cisplatin. RES pretreatment for 1 h with 10 or 15 uM RES increased HK‐2 viability relative to cells exposed only to RES Vehicle (DMSO). Additional studies examined whether RES stimulated cell growth as part of the mechanism for cellular protection. RES did not increase cell number as part of its protective mechanism. ATP levels were diminished by cisplatin causing an increase in the relative balance of ADP/ATP. RES (5 and 7.5 uM) did not alter ATP or ADP/ATP ratios. Additional studies examined Caspase cleavage, protein carbonylation and 4‐hydroxynonenal adduction (4‐HNE) following cisplatin exposure of HK‐2 cells pretreated with DMSO or RES. Western analysis detected an increase in protein carbonylation with 24 h cisplatin exposure which was reversed by RES. In summary, RES protects HK‐2 cells from cisplatin cytotoxicity and preserves mitochondrial ATP levels. Support or Funding Information (Supported by NIH Grants INBRE 3P20RR016477‐09S4; 5P20RR016477 and 8P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence).