Stress-Specific Activation of Heat Shock Factor (HSF) and Hypoxia-Inducible Factor (HIF) in the Kidney

Abstract 1950 Nephrology Platform, Sunday, 5/2

Ischemic renal injury consists of at least two major stresses: i) a decrease of oxygen (O₂) supply (hypoxia) and ii) limited availability of cellular energy (ATP depletion). Each of these injuries is believed to rapidly activate stress-sensing transcription factors. Therefore, in LLC-PK₁ cells in vitro and ischemic rat kidneys in vivo, we investigated activation of two transcription factors: HSF which is known to induce production of a variety of heat-shock proteins and HIF which orchestrates and modulates the molecular response to hypoxia. Cells were grown to confluence, serum deprived for 24 hours, and then subjected to either hypoxia or substrate depletion. Hypoxia was achieved by incubation in 0.1% - 5% O₂. Substrate depletion occurred when Antimycin-A (a mitochondrial inhibitor) was added to serum and glucose-free media. HSF and HIF activation were analyzed by gel shift assays using nuclear extracts incubated with radiolabeled oligonucleotides comprising the HSE (heat shock element) or HRE (hypoxia-response element), respectively.

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