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Zongbo WEI, Vincent M Monnier, Xingjun Fan; Lens GSH deficiency mouse Promote Alpha-smooth muscle actin expression. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2022.
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© ARVO (1962-2015); The Authors (2016-present)
The anterior subcapsular cataract and posterior capsular opacification (PCO) are contributed by lens epithelial cell proliferation, migration, and transdifferentiation. One of the key characteristic features of epithelial cell transdifferentiation and myofiborblast formation is the expression of alph smooth muscle actin (aSMA). The lens maintains unusually high level of glutathione (GSH), while no GSH is present in intro-ocular lens (IOL). We hypothesis that residual lens epithelial cells lacking interaction with lens GSH may promote its transdifferentiation and myofiborblast formation.
Wild type and lens conditional gamma glutamyl cysteine ligase, catalytic subunit (Gclc) knockout mouse (LEGSKO mouse) lenses were extracted, and the aSMA was determined by Western-blot analysis using aSMA antibody. The potential involvement of autophagy was also characterized by determining the LC3 I/II protein level via Western-blot analysis. The aSMA and LC3I/II were also determined in human lens epithelial cells (HLE-B3), mouse lens epithelial cells (17EM15), and primary mouse lens epithelial cells after Gclc knocking down by shRNAlenti-virus. Immunohistochemistry was also used to detect and quantitate alpha-SMA expression.
Preliminary data indicated that there was at least a three fold of aSMA elevation in LEGSKO mouse lens compared to wild type mouse (p<0.05). In contrast, the autophagy marker LC3 was reduced significantly in LEGSKO lens compared to wild type mouse.
These results implicate the important role of redox status in residual epithelial cells transdifferentiation after cataract surgery.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
a-SMA was increased in LEGSKO mice.
LC3 was decreased in LEGSKO mice
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