Oxidative stress-induced alterations in PPAR-γ and associated mitochondrial destabilization contribute to kidney cell apoptosis

The mechanism(s) underlying renoprotection by peroxisome proliferator-activated receptor (PPAR)-gamma agonists in diabetic and nondiabetic kidney disease are not well understood. Mitochondrial dysfunction and oxidative stress contribute to kidney disease. PPAR-gamma upregulates proteins required for mitochondrial biogenesis. Our aim was to determine whether PPAR-gamma has a role in protecting the kidney proximal tubular epithelium (PTE) against mitochondrial destabilisation and oxidative stress. HK-2 PTE cells were subjected to oxidative stress (0.2-1.0 mM H2O2) for 2 and 18 h and compared with untreated cells for apoptosis, mitosis (morphology/biomarkers), cell viability (MTT), superoxide (dihydroethidium), mitochondrial function (MitoTracker red and JC-1), ATP (luminescence), and mitochondrial ultrastructure. PPAR-gamma, phospho-PPAR-gamma, PPAR-gamma coactivator (PGC)-1 alpha, Parkin (Park2), p62, and light chain (LC)3 beta were investigated using Western blots. PPAR-gamma was modulated using the agonists rosiglitazone, pioglitazone, and troglitazone. Mitochondrial destabilization increased with H2O2 concentration, ATP decreased (2 and 18 h; P < 0.05), Mitotracker red and JC-1 fluorescence indicated loss of mitochondrial membrane potential, and superoxide increased (18 h, P < 0.05). Electron microscopy indicated sparse mitochondria, with disrupted cristae. Mitophagy was evident at 2 h (Park2 and LC3 beta increased; p62 decreased). Impaired mitophagy was indicated by p62 accumulation at 18 h (P < 0.05). PPAR-gamma expression decreased, phospho-PPAR-gamma increased, and PGC-1 alpha decreased (2 h), indicating aberrant PPAR-gamma activation and reduced mitochondrial biogenesis. Cell viability decreased (2 and 18 h, P < 0.05). PPAR-gamma agonists promoted further apoptosis. In summary, oxidative stress promoted mitochondrial destabilisation in kidney PTE, in association with increased PPAR-gamma phosphorylation. PPAR-gamma agonists failed to protect PTE. Despite positive effects in other tissues, PPAR-gamma activation appears to be detrimental to kidney PTE health when oxidative stress induces damage.