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  • Title: Connexin43 knockdown accelerates wound healing but inhibits mesenchymal transition after corneal endothelial injury in vivo.
    Author: Nakano Y, Oyamada M, Dai P, Nakagami T, Kinoshita S, Takamatsu T.
    Journal: Invest Ophthalmol Vis Sci; 2008 Jan; 49(1):93-104. PubMed ID: 18172080.
    Abstract:
    PURPOSE: To explore connexin43 (Cx43) knockdown as an efficient treatment for corneal endothelial injury in an in vivo rat corneal scrape injury model. METHODS: Scrape injury was induced in the corneal endothelium, and immunolabeling (ZO-1, alpha-SMA, Cx43) was performed to analyze changes in Cx43 expression during wound healing. Single injection of Cx43 antisense oligodeoxynucleotide (AS-ODN), small interfering RNA (siRNA), or adenovirus (CMV-Cx43-mRFP1) was applied into the anterior chamber simultaneously with the injury, and wound closure was examined by immunolabeling (ZO-1, Cx43) and propidium iodide staining. Corneal endothelium proliferation on day 1 after injury was studied by Ki67-immunolabeling. Cx43-knockdown treatment was performed also without injury, and its effect on Cx43 expression and Ki67 immunolabeling was examined. The postinjury appearance of myofibroblasts in Cx43 AS-ODN- and sense-ODN-treated corneas was compared by alpha-SMA-immunolabeling. RESULTS: Complete wound closures were observed in five of six corneas on day 3 after injury with either Cx43 AS-ODN or siRNA treatment, whereas no complete closure was observed on day 3 in the control corneas (S-ODN, zero of six; or nonsense siRNA, zero of six). Consistently, Cx43 overexpression using adenovirus delayed wound closure. Cx43 knockdown increased the number of Ki67-positive proliferating cells on day 1, whereas it decreased the number of alpha-SMA-positive myofibroblasts on day 5. Cx43 knockdown without injury decreased Cx43 expression and induced endothelial proliferation in vivo. CONCLUSIONS: These results show that Cx43 knockdown induces corneal endothelium proliferation but inhibits endothelial-mesenchymal transition/transformation after injury, suggesting that Cx43 knockdown is a new therapeutic approach for acceleration of wound closure and for prevention of retrocorneal fibrous membrane formation.
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