Inomata Takenori, Fujimoto Keiichi, Okumura Yuichi, Zhu Jun, Fujio Kenta, Shokirova Hurramhon, Miura Maria, Okano Mikiko, Funaki Toshinari, Sung Jaemyoung, Negishi Naoko, Murakami Akira
Department of Ophthalmology, Juntendo University Graduate School of Medicine, 3-1-3 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
Department of Ophthalmology, Juntendo University Faculty of Medicine, Tokyo, Japan.
Sci Rep. 2020 Nov 13;10(1):19817. doi: 10.1038/s41598-020-76882-w.
Corneal allograft survival is mediated by the variety of immunological reactions and wound healing process. Our aim was to explore the effects of topical administration of ripasudil, a selective Rho-associated coiled-coil protein kinase inhibitor, on corneal allograft survival. Ripasudil was administered to mice thrice a day after allogeneic corneal transplantation. Corneal graft survival, opacity, neovascularization, re-epithelization, immune cell infiltration, and mRNA levels of angiogenic and pro-inflammatory factors in the grafted cornea and draining lymph nodes (dLNs) were evaluated with slit-lamp microscopy, immunohistochemistry, flow cytometry, and polymerase chain reaction. Graft survival was significantly prolonged with lower graft opacity and neovascularization scores in 0.4% and 2.0% ripasudil-treated groups, and mRNA levels of angiogenic and pro-inflammatory factors in ripasudil-treated grafted corneas were reduced. Moreover, 0.4% and 2.0% ripasudil reduced CD45-infiltrated leukocyte frequency, Cd11b and Cd11c mRNA levels, and the frequencies of mature dendritic cells, IFNγ-, and IL-17- producing CD4T cells in the dLNs of recipients. Re-epithelization rate of the grafted cornea was significantly higher in the 0.4% and 2.0% ripasudil groups than in the control. Topically applied ripasudil prolonged graft survival by downregulating neovascularization and inflammation factors, while promoting corneal re-epithelization, suggesting that ripasudil may be useful for suppressing immunological rejection in corneal transplantation.
同种异体角膜移植的存活受多种免疫反应和伤口愈合过程的介导。我们的目的是探讨局部应用选择性Rho相关卷曲螺旋蛋白激酶抑制剂ripasudil对同种异体角膜移植存活的影响。在同种异体角膜移植后，ripasudil每天给药小鼠三次。通过裂隙灯显微镜检查、免疫组织化学、流式细胞术和聚合酶链反应评估角膜移植的存活、混浊、新生血管形成、再上皮化、免疫细胞浸润以及移植角膜和引流淋巴结（dLNs）中血管生成和促炎因子的mRNA水平。在0.4%和2.0%的ripasudil治疗组中，移植存活时间显著延长，移植混浊和新生血管形成评分较低，ripasudil治疗的移植角膜中血管生成和促炎因子的mRNA水平降低。此外，0.4%和2.0%的ripasudil降低了受体dLNs中CD45浸润白细胞频率、Cd11b和Cd11c mRNA水平以及成熟树突状细胞、产生IFNγ和IL-17的CD4T细胞频率。0.4%和2.0%的ripasudil组移植角膜的再上皮化率显著高于对照组。局部应用ripasudil通过下调新生血管形成和炎症因子来延长移植存活时间，同时促进角膜再上皮化，这表明ripasudil可能有助于抑制角膜移植中的免疫排斥反应。
