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Sirt3 regulates mitophagy level to promote diabetic corneal epithelial wound healing.
Experimental Eye Research 2019 Februrary 20
We aim to investigate how Sirt3 (silent mating type information regulation 2 homolog 3) promoting diabetic corneal epithelial wound healing by regulating mitophagy. The effect of HG(High Glucose, 25 mM D-glucose) on Sirt3 and LC3B(light chain 3 beta)which representing of mitophagy were investigated in TKE2 cells (a murine limbal/corneal epithelium-derived progenitor cell line) and corneal epithelium from C57BL/6J-Ins2Akita (Ins2Akita/+ ) mice using RT-PCR and Western blotting. How overexpression of Sirt3 promoting diabetic corneal epithelial wound healing was investigated with cell migration assay、immunofluorescence、 immunofluorescence colocalization and corneal injury model. We found that HG reduced the expression of Sirt3 as well the mitophagy both in TKE2 cells and corneas from Ins2Akita/+ mice. And overexpression of Sirt3 prominently promoted wound healing speed under HG condition via upregulating the level of mitophagy. Mitophagy level was increased dramatically when the Foxo3a (Forkhead box O3)/PINK1(PTEN Induced putative kinase protein 1)-Parkin pathway was activated by Sirt3 overexpression which suggested that the mitophagy was involved in cell injury under HG condition. This study demonstrated the mechanism of Sirt3 regulating mitophagy to promote diabetic corneal epithelial wound healing in vivo and in vitro, which suggested that Sirt3 may positively impact diabetic keratopathy(DK).
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