P66Shc expression in diabetic rat retina

BMC Ophthalmol. 2018 Feb 27;18(1):58. doi: 10.1186/s12886-018-0724-3.

Abstract

Background: P66Shc is partially localised within the mitochondrial fraction. It is primarily related to the generation of mitochondrial reactive oxygen species and apoptosis. Based on previous studies, we hypothesize that in the retina, p66Shc may exist and affect the development of diabetic retinopathy. The purpose of this study was to investigate p66Shc expression in retinal in streptozotocin-induced diabetic (SD) rats, which may provide a pathway to study the pathogenesis of diabetic retinopathy.

Methods: Reverse transcription-polymerase chain reaction (RT-PCR) and western blot were used to detect retinal p66Shc mRNA and protein expression in SD rats, respectively. Immunohistochemical staining was applied to detect the location of rat retinal p66Shc expression. TUNEL assay was applied to detect the number of apoptotic cells.

Results: P66Shc expression was found in the retina of normal and diabetic rats, and the level of mRNA and protein expression increased with the progression of diabetes mellitus (DM). P66Shc expression was mainly located in the retinal ganglion cell layer and inner nuclear layer. Compared with the normal group, retinal cell tissue apoptosis rate in the D12w group was significantly increased.

Conclusion: Rat retinal p66Shc expression was mainly in the ganglion cell layer and inner nuclear layer. As the degree of DM progressed, p66Shc expression gradually increased, and the number of apoptotic cells also increased.

Keywords: Apoptosis; Diabetic retinopathy; Oxidative stress; p66Shc, reactive oxygen species.

MeSH terms

  • Animals
  • Blotting, Western
  • Diabetes Mellitus, Experimental / metabolism*
  • Diabetic Retinopathy / metabolism*
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Male
  • Rats
  • Rats, Sprague-Dawley
  • Real-Time Polymerase Chain Reaction
  • Retina / metabolism*
  • Retinal Ganglion Cells / metabolism
  • Src Homology 2 Domain-Containing, Transforming Protein 1 / metabolism*

Substances

  • Shc1 protein, rat
  • Src Homology 2 Domain-Containing, Transforming Protein 1