Rho-associated protein kinase inhibitor, Y-27632, significantly enhances cell adhesion and induces a delay in G1 to S phase transition in rabbit corneal endothelial cells
Rho-associated protein kinase inhibitor, Y-27632, significantly enhances cell adhesion and induces a delay in G1 to S phase transition in rabbit corneal endothelial cells
复制标题
DOI：
10.3892/mmr.2015.3628
复制
发表时间：
2015-08-01
影响因子：
3.4
通讯作者：
Hong, Jing
中科院分区：
文献类型：
--
作者：
Diao, Yu-Mei;Hong, Jing
人角膜内皮细胞是一种非增殖性细胞类型。由于角膜内皮疾病的增加，已经进行了越来越多的研究以促进角膜内皮细胞增殖。本研究的目的是研究Rho相关蛋白激酶抑制剂Y-27632对兔角膜内皮细胞（rCEC）增殖的影响。在两个不同的时间点将Y-27632（1、10或30 μ M）加入到两组rCEC中。第一组在rCEC最初接种时接受Y-27632，第二组在细胞生长72小时后接受Y-27632。随后使用光学显微镜观察细胞形态和细胞粘附率。使用细胞计数试剂盒测量粘附于培养板的活细胞的数量。流式细胞仪检测细胞周期转换和Annexin V阳性凋亡细胞的水平。用1 μ M Y-27632和10 μ M Y-27632处理的细胞保持其细胞形状。在30 μ M Y-27632的浓度下，细胞形状变得不规则。1 μ M Y-27632、10 μ M Y-27632和30 μ M Y-27632的细胞粘附率分别为36.84%、84.21%和84.21%，均高于对照组（P
Human corneal endothelial cells are a non-prolif-erative cell type. As a result of the increase in corneal endothelium disease, increasing numbers of studies have been conducted in order to promote corneal endothelial cell proliferation. The aim of the present study was to investigate the proliferative effects of Rho-associated protein kinase inhibitor, Y-27632, on rabbit corneal endothelial cells (rCECs). Y-27632 (1, 10 or 30 mu M) was added at two different time points to two groups of rCECs. The first group received Y-27632 when rCECs were initially plated, and the second following 72 h of cell growth. Cell morphology and cell adhesion ratios were subsequently observed using light microscopy. A cell counting kit was used to measure the number of viable cells that adhered to culture plates. Cell cycle transitions and levels of Annexin V-positive apoptotic cells were detected using flow cytometry. Cells treated with 1 mu M Y-27632 and 10 mu M Y-27632 retained their cell shape. At a concentration of 30 mu M Y-27632, the cell shape became irregular. Cell adhesion ratios, in 1 mu M Y-27632 (36.84%), 10 mu M Y-27632 (84.21%) and 30 mu M Y-27632 (84.21%) were higher than the adhesion ratio in the control group (P