1. From: Overexpression of SIRT1 Promotes High Glucose–Attenuated Corneal Epithelial Wound Healing via p53 Regulation of the IGFBP3/IGF-1R/AKT Pathway
Invest. Ophthalmol. Vis. Sci ;54(5): doi: /iovs
Figure Legend:
The overexpression of SIRT1 promotes HG attenuation of corneal epithelial wound healing via regulation of the IGFBP3/IGF-1R/AKT pathway by p53 in vivo. (A) Ectopic expression of SIRT1 on Akita mouse corneal epithelia. After subconjunctival injection, SIRT1 was localized in the corneal epithelium. Compared with the corneal epithelia of the noninfected group (Ai), the corneal epithelia of the AD-SIRT1–infected group were characterized by high SIRT1 expression (Aii). (B) The effect of SIRT1 overexpression on corneal wound healing in Ins2Akita/+ mice. Left panels: the fluorescein-stained corneas at 24 and 48 hours after injury. Right panel: the remaining wound area at 24 and 48 hours after injury. To further investigate whether SIRT1 is involved in the pathogenesis of diabetic keratopathy, we examined the effects of ectopic SIRT1 expression on the corneal epithelial debridement wound healing using type 1 diabetic Ins2Akita/+ mice and control Ins2+/+ mice. Either AD-SIRT1 or AD-GFP viral preparations were injected into the subconjunctival site on the same day of corneal epithelium injury, and the corneal surface wound was monitored using fluorescein dye. At 48 hours postinjury, the wound area was significantly reduced in the mice administered AD-SIRT1 compared with the saline-treated or AD-GFP–infected groups. The image shows the remaining wound area at 24 and 48 hours after injury. Changes in the mean remaining wound areas in pixels were calculated using commercial editing software (*P < 0.05). (C) The effect of regulation of the IGFBP3/IGF-1/AKT pathway by p53 on corneal wound healing in mice after SIRT1 overexpression. Left panels: the representative gel results. Right panel: the graphs used for normalization. At 48 hours postinjury the corneas were harvested and derived lysates were subjected to Western blotting by using anti-SIRT1, anti-acety-p53 (K379), anti-p53, anti-IGFBP3, anti-IGF-1R, anti-p-AKT, or anti-AKT. GAPDH served as the loading control. The injured but untreated Akita mice cornea served as the control. Data are means ± SD (n = 3). Significant differences (*P < 0.05) between the control and AD-SIRT1 groups are identified by “*.”
Date of download: 1/3/2018
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