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. 2021 Apr 21;11:8647. doi: 10.1038/s41598-021-88118-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Inhibitory effects of topically administered nalfurafine on the expression of angiogenic factors in the neovascularized cornea. Corneal neovascularization was induced by placing intrastromal figure-of-eight suture knots into the central cornea of BALB/c mice for 14 days. Eye drops containing the kappa opioid receptor agonist nalfurafine were administered topically (0.1 µg/2 μL/eye) once or twice a day to the neovascularized cornea for 14 days. Significant increase in the mRNA expression level of Oprk1 encoding kappa opioid receptor 1 was observed, 14 days post treatment in the nalfurafine twice a day samples (a). Relative mRNA expression levels of angiogenic factors in the control and treated neovascularized corneal samples were analyzed using RT-qPCR (b–g). Significant decreases in mRNA levels were observed,14 days post treatment in samples that were exposed to nalfurafine once and/or twice a day for the following VEGF signaling components: Vegfa (b), Vegfc (c), Flt1 (d), Kdr (e), Flt4 (f), and Nrp1 (g). Data are presented as the mean ± standard error of the mean (n = 3 in all cases). Statistical significance of differences is illustrated as follows: *P < 0.05; **P < 0.01; ***P < 0.001 (one-way analysis of variance followed by the Bonferroni test). (h) Heatmap shows relative decreases in expression levels of angiogenic factors in the neovascularized cornea depending on the dosage of nalfurafine. ns, no significant difference.