From the Department of Ophthalmology, Fujian Medical University Union Hospital, Fu Zhou, China.
From the Department of Ophthalmology, Fujian Medical University Union Hospital, Fu Zhou, China.
Exp Eye Res. 2019 Apr;181:223-231. doi: 10.1016/j.exer.2019.02.011. Epub 2019 Feb 19.
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 (Ins2) 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 Ins2 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).
我们旨在研究 Sirt3（沉默交配型信息调节 2 同源物 3）如何通过调节线粒体自噬来促进糖尿病性角膜上皮伤口愈合。使用 RT-PCR 和 Western blot 法在 TKE2 细胞（一种鼠角膜缘/上皮衍生祖细胞系）和 C57BL/6J-Ins2Akita（Ins2）小鼠角膜中研究了高糖（25 mM D-葡萄糖）对 Sirt3 和代表线粒体自噬的 LC3B（微管相关蛋白轻链 3B）的影响。通过细胞迁移测定、免疫荧光、免疫荧光共定位和角膜损伤模型研究了 Sirt3 过表达如何促进糖尿病性角膜上皮伤口愈合。我们发现，HG 降低了 TKE2 细胞和 Ins2 小鼠角膜中 Sirt3 的表达以及线粒体自噬水平。并且，Sirt3 的过表达显著促进了 HG 条件下的伤口愈合速度，通过上调线粒体自噬水平。当 Sirt3 过表达激活 Foxo3a（叉头框 O3）/PINK1（PTEN 诱导的假定激酶蛋白 1）-Parkin 通路时，线粒体自噬水平显着增加，这表明线粒体自噬参与了 HG 条件下的细胞损伤。这项研究证明了 Sirt3 通过调节线粒体自噬来促进体内和体外糖尿病性角膜上皮伤口愈合的机制，表明 Sirt3 可能对糖尿病性角膜病变（DK）产生积极影响。
