Abstract：
Penetrating keratoplasty remains the most common treatment to restore vision for corneal diseases. Immune rejection after corneal transplantation is one of the major causes of graft failure. In recent years, Rho-associated protein kinase (ROCK) inhibitors have been found to be associated with the activation of the STATs pathway and are widely studied in autoimmune diseases. Therefore, it may be possible that the ROCK inhibitors also participate in the local and systemic immune regulation in corneal transplantation through activation of the STATs pathway and affect the CD4+ T cell differentiation. This study aimed to explore the role of ROCK-STATs pathway in the occurrence of immune rejection in corneal transplantation by applying Y27632, a ROCK inhibitor, to the recipient mice and peripheral CD4+ T cells. We found that Y27632 significantly up-regulated the phosphorylation level of STAT5 in both spleen and lymph nodes, down-regulated the phosphorylation level of STAT3 in the CD4+ T cells in the spleen. It also increased the proportion of CD4+CD25+Foxp3+Helios+ Tregs while decreased CD4+IL17A+ -Th17 cells. Moreover, Y27632 also reduced the proportion of dendritic cells in both spleen and lymph nodes, as well as the expression level of CD86 on their surfaces in the spleen, while the proportion of macrophages was not affected. The expression levels of ROCK1, ROCK2, CD11c and IL-17A mRNA were also found to be low in the graft tissue while the expression of Helios was upregulated. Rho-kinase inhibitor can modulate the balance of Tregs/Th17 by regulating the phosphorylation levels of both STAT3 and STAT5, thereby inhibiting the occurrence of immune rejection in allogeneic corneal transplantation.
摘要：
穿透性角膜移植术仍然是恢复角膜疾病视力的最常见治疗方法。角膜移植后的免疫排斥反应是导致移植失败的主要原因之一。近年来,已发现Rho相关蛋白激酶（ROCK）抑制剂与STATs途径的激活有关，并且在自身免疫性疾病中被广泛研究。因此,ROCK抑制剂也可能通过激活STATs通路参与角膜移植的局部和全身免疫调节,影响CD4+T细胞分化。本研究通过应用ROCK抑制剂Y27632,探讨ROCK-STATs通路在角膜移植免疫排斥反应发生中的作用,给受体小鼠和外周CD4+T细胞。我们发现Y27632显著上调脾脏和淋巴结中STAT5的磷酸化水平,下调脾脏CD4+T细胞中STAT3的磷酸化水平。它还增加了CD4+CD25+Foxp3+Helios+Tregs的比例，同时减少了CD4+IL17A+-Th17细胞。此外，Y27632还降低了脾脏和淋巴结中树突状细胞的比例，以及CD86在脾脏表面的表达水平，而巨噬细胞的比例没有受到影响。在移植组织中，ROCK1，ROCK2，CD11c和IL-17AmRNA的表达水平也较低，而Helios的表达上调。Rho激酶抑制剂可以通过调节STAT3和STAT5的磷酸化水平来调节Tregs/Th17的平衡，从而抑制同种异体角膜移植中免疫排斥的发生。
