ZY-27632 is an orally potent, ATP-competitive inhibitor of ROCK-I and ROCK-II. Y-27632 also inhibits isolation-induced apoptosis in mouse prostate stem or progenitor cells.

ROCKI:220 nM(ki), ROCKII:300 nM(ki)

方法：人诱导多能干细胞 marmoset iPSC 用 Y-27632 (5-20 μM) 处理 7 天，使用 AKP 检测克隆形成情况。
结果：Y-27632 显著提高 marmoset iPSC 的克隆效率。[1]
方法：成人脂肪组织衍生干细胞 ADSCs 用 Y-27632 (5 μmol/L) 处理 1 h，检测 ADSCs 的形态变化。
结果：Y-27632 剂量依赖性诱导 ADSCs 的神经元分化，5 μmol/L Y-27632 处理 1 h 的 ADSCs的神经元样细胞百分比为（93.5±4.7）%。[2]
方法：食蟹猴胚胎干细胞 cyES 常规传代或用 Y-27632 (1-10 μM) 处理 24 h，使用 Flow Cytometry 方法进行活-死染色，使用试剂盒检测 BrdU。
结果：Y-27632 促进 cyES 存活细胞增加。Y-27632 没有促进细胞增殖，但保护细胞在单细胞消化后免于细胞死亡。[3]

方法：为研究 Y-27632 在运动神经元疾病的治疗潜力，将 Y-27632 (2 or 30 mg/kg in drinking water) 口服给 ALS 模型的 SOD1G93A 小鼠，持续 137 天。
结果：Y-27632 2 mg/kg 治疗的效果不佳，Y-27632 30 mg/kg 治疗可改善雄性小鼠的运动功能，雌性小鼠仅表现出有限的改善。[4]
方法：为研究 Y-27632 对肝纤维化的影响，将 Y-27632 (30 mg/kg) 口服给药给 dimethylnitrosamine (DMN) 诱导肝纤维化的大鼠，每天一次，持续四周。
结果：Y-27632 治疗显著减少了 DMN 诱导的肝纤维化的发生，并降低了肝脏中胶原和羟脯氨酸的含量以及 α-SMA 的表达。[5]

Recombinant ROCK-I, ROCK-II, PKN, or citron kinase is expressed in HeLa cells as Myc-tagged proteins by transfection using Lipofectamine, and is precipitated from the cell lysates by the use of 9E10 monoclonal anti-Myc antibody coupled to G protein-Sepharose. Recovered immunocomplexes are incubated with various concentrations of [32P]ATP and 10 mg of histone type 2 as substrates in the absence or presence of various concentrations of either Y-27632 or Y-30141 at 30°C for 30 min in a total volume of 30 μL of the kinase buffer containing 50 mM HEPES-NaOH, pH 7.4, 10 mM MgCl2, 5 mM MnCl2, 0.02% Briji 35, and 2 mM dithiothreitol. PKCa is incubated with 5 μM [32P]ATP and 200 μg/mL histone type 2 as substrates in the absence or presence of various concentrations of either Y-27632 or Y-30141 at 30°C for 10 min in a kinase buffer containing 50 mM Tris-HCl, pH 7.5, 0.5 mM CaCl2, 5 mM magnesium acetate, 25 μg/mL phosphatidyl serine, 50 ng/mL 12-O-tetradecanoylphorbol-13-acetate and 0.001% leupeptin in a total volume of 30 μL. Incubation is terminated by the addition of 10 μL of 43 Laemmli sample buffer. After boiling for 5 min, the mixture is subjected to SDS-polyacrylamide gel electrophoresis on a 16% gel. The gel is stained with Coomassie Brilliant Blue, and then dried. The bands corresponding to histone type 2 are excised, and the radioactivity is measured[1].

Y-27632 is dissolved in water and stored[1]. HeLa cells are plated at a density of 3×104 cells per 3.5-cm dish. The cells are cultured in DMEM containing 10% FBS in the presence of 10 mM Thymidine for 16 h. After the cells are washed with DMEM containing 10% FBS, they are cultured for an additional 8 h, and then 40 ng/mL of Nocodazole is added. After 11.5 h of the Nocodazole treatment, various concentrations of Y-27632 (0-300 μM), Y-30141, or vehicle is added and the cells are incubated for another 30 min[1].