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Resveratrol Protection of Mitochondrial Function in HK‐2 Cells Exposed to Cisplatin.
Author(s) -
Valentovic Monica,
Brown Katie,
Lamyaithong Andre Benja,
Dial Mason
Publication year - 2017
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.31.1_supplement.819.6
Subject(s) - cisplatin , viability assay , chemistry , oxidative stress , mtt assay , reactive oxygen species , pharmacology , resveratrol , cytotoxicity , mitochondrion , apoptosis , microbiology and biotechnology , biochemistry , biology , medicine , chemotherapy , in vitro
Cisplatin is a cancer chemotherapeutic agent widely used to control cancer to the testes, ovaries and cervix. Cisplatin is nephrotoxic and with a potential incidence of 33%. Resveratrol (RES) is a polyphenolic compound that possesses anticancer activity. Part of the mechanism for reducing cisplatin toxicity by RES may be mediated by preserving mitochondrial function and diminishing cisplatin induced oxidative stress. Our hypothesis is that RES will prevent mitochondrial impairment by cisplatin in HK‐2 cells. HK‐2 cells were selected as they are a human proximal tubular kidney cell line (HK‐2). Hk‐2 cells were plated for 48 followed by a 1 h pretreated with RES or 1% DMSO (vehicle). Renal cells were subsequently exposed to cisplatin at a final concentration of 0–50 uM for 24 h. Viability was assessed using MTT release and cell count with an n=6/group and repeated 3 separate times. Cells pretreated with 10 uM RES were protected from cisplatin cytotoxicity at 24 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). Mitochondrial respiration was measured using a Seahorse XFp analyzer and expressed as oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Mitochondrial function was further probed by monitoring ATP levels and cytochrome c leakage. Basal and uncoupled (FCCP) mitochondrial respiration were monitored in RES and cisplatin treated cells. Additional studies examined oxidative stress as increased 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. Mitochondrial function was diminished by cisplatin and protected by RES. RES protects human proximal tubular epithelial cells from cisplatin cytotoxicity, preserves mitochondrial integrity and mitochondrial respiration. Support or Funding Information (Supported by NIH Grants INBRE 3P20RR016477‐09S4; 5P20RR016477 and 8P20GM103434 and a WV NASA Undergraduate Research Fellowship to MD.