title: Metabolic stress response implicated in diabetic retinopathy: the role of calpain, and the therapeutic impact of calpain inhibitor.
abstract: To describe how a high fat diet (HFD) and hyperglycemia initiate a sequence of calpain activation and oxidative stress associated with neuro-degenerative changes in diabetic retinopathy (DR), hyperglycemia was induced with streptozotocin in mice lacking the gene for calpastatin (CAST KO), and in mice lacking the gene for the transcription factor NF-E2 related factor 2 (Nrf2 KO). All animals were fed a HFD. Retinal ganglion cell (RGC) density was estimated by labeling with fluorogold and immunohistochemistry. A potent calpain inhibitor, SNJ-1945, was administered daily until the animals were sacrificed. In vitro, oxidative stress-induced RGC loss was evaluated in a high glucose culture medium with and without SNJ-1945. Retinal mRNA of calpain-1 and calpain-2 was measured by quantitative RT-PCR. Pre-apoptotic substrates of cleaved α-fodrin and synaptophysin protein were quantified by immunoblot analysis. Axonal damage was examined in transverse sections of the optic nerve. A HFD and hyperglycemia significantly increased RGC and axonal degeneration 3 weeks into the experiment. Levels of cleaved α-fodrin were increased. In the CAST KO mice, the neurotoxicity was augmented significantly. Gene manipulation of CAST and orally administered SNJ-1945 successfully modified calpain levels in the retina and prevented RGC death. In vitro, a high-glucose culture of retinal cells without antioxidants showed more RGC death than that with antioxidant treatment. The expression of synaptophysin was significantly suppressed by SNJ-1945 treatment. These results suggest that calpain plays a crucial role in metabolic-induced RGC degeneration caused by hyperglycemia and oxidative stress. Antioxidant and calpain inhibition offers important opportunities for future neuroprotective treatment against RGC death in various metabolic stress-induced diseases including DR.
authors.affiliation: Department of Ophthalmology, Tohoku University Graduate School of Medicine, Miyagi, Japan.
authors.forename: Ahmed Y
authors.initials: AY
authors.lastname: Shanab
authors.forename: Toru
authors.initials: T
authors.lastname: Nakazawa
authors.forename: Morin
authors.initials: M
authors.lastname: Ryu
authors.forename: Yuji
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Noriko
authors.initials: N
authors.lastname: Himori
authors.forename: Keiko
authors.initials: K
authors.lastname: Taguchi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yasuda
authors.forename: Ryo
authors.initials: R
authors.lastname: Watanabe
authors.forename: Jiro
authors.initials: J
authors.lastname: Takano
authors.forename: Takaomi
authors.initials: T
authors.lastname: Saido
authors.forename: Naoko
authors.initials: N
authors.lastname: Minegishi
authors.forename: Toshio
authors.initials: T
authors.lastname: Miyata
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Abe
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D002154:Calpain / Q000378:metabolism*; D003921:Diabetes Mellitus, Experimental / Q000150:complications; D003930:Diabetic Retinopathy / Q000378:metabolism*; D059305:Diet, High-Fat / Q000009:adverse effects; D006023:Glycoproteins / Q000494:pharmacology*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013312:Stress, Physiological / Q000187:drug effects / Q000502:physiology*
chemical_list: D006023:Glycoproteins; C039304:calpain inhibitors; D002154:Calpain