A Novel Specific Application of Pyruvate Protects the Mouse Retina against White Light Damage: Differential Stabilization of HIF-1α and HIF-2α

PURPOSE. To mimic hypoxia preconditioning by a novel specific pyruvate treatment and to study its retinal protection against white light damage. METHODS. Six-to-eight-week-old BALB/c mice were exposed to strong white light calculated to produce photoreceptor degeneration. Some were given injections of pyruvate in a preordained protocol because evidence exists that proves pyruvate can affect the concentration of hypoxia inducible factor (HIF). Western blotting and real-time PCR were used to determine the concentration of proteins and mRNAs in retinas. Morphology was analyzed with toluidine blue staining and was plotted using a spidergraph. A free nucleosome cell death assay was used to examine apoptosis. Retina explant cultures were used to investigate the background mechanism. RESULTS. Pyruvate administration stabilized hypoxia inducible factor (HIF)-1 alpha but not HIF-2 alpha. Expression of the downstream genes hemoxygenase-1 and erythropoietin mirrored the changes of the two HIFs, respectively. Importantly, pyruvate given not only before but also after exposure to light protected photoreceptors against apoptosis. In the retinal explant system, addition or depletion of pyruvate caused only changes of HIF-1 alpha and prolyl hydroxylase (PHD)-2, while HIF-2 alpha and PHD1 were not affected. However, under hypoxic conditions, HIF-2 alpha was stabilized by pyruvate but not HIF-1 alpha. CONCLUSIONS. Pyruvate evoked a hypoxia-like response under normoxic conditions and was retina-protective against strong white light. This response included stabilization of HIF-1 alpha but not HIF-2 alpha. This differential stabilization might be related to the distinct preference of their degrading enzyme of PHD2 and PHD1 in response to pyruvate treatment. (Invest Ophthalmol Vis Sci. 2011;52:3112-3118) DOI:10.1167/iovs.10-5605