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Subrata k. das, Jr., Ling Luo, Nirbhai Singh, Hironori Uehara, Bonnie Archer, Jacquelyn M. Simonis, Ying Liu, Thomas Olsen, Balamurali K. Ambati; Role Of Vimentin In Corneal Fibrosis And Wound Healing Process. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6382.
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Vimentin is an intermediate filament protein that contributes to filamentous network between the cell periphery and the nucleus and it has a dynamic link between the nucleus and extracellular matrix. In addition, it is thought to be involved in structural processes such as wound healing. Our main purpose is to examine the role of vimentin for ocular fibrosis at the stage of wound healing.
C57BL6/j mice corneas were sutured (3 sutures) to induce corneal angiogenesis. Immunohistochemical double staining of α-smooth muscle actin (α-SMA) or vimentin or glial fibrillary acidic protein (GFAP) and CD31 was performed to determine the expression level of vimentin, GFAP and α-SMA in neovascularized and normal corneas 10 days after suture. We designed three distinct plasmids expressing shRNA targeting vimentin mRNA of both mice and humans. Cultured human umbilical vein endothelial cells (HUVEC) were transfected by vimentin shRNA plasmids and the same concentration of the plasmid coding nonspecific shRNA was used as control. The transfection efficiency was examined using GFP-expressing plasmids. Vimentin mRNA expression was evaluated by semi-quantitative RT-PCR.
: Immunostaining indicated that vimentin and GFAP increased in neovascularized (NV) cornea compared to normal cornea 10 days post-suture. Only α-SMA was detected in vessels of neovascularized cornea but not in normal cornea. We found that all the three vimentin shRNA plasmids decrease the expression level of vimentin compared to the control in HUVEC.
Vimentin and GFAP expression are correlated with fibrosis during wound healing. Vimentin shRNA knockdown have potential to be developed as anti-fibrosis treatments.
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