| 名稱 | Y-27632 |
| 描述 | 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. |
| 細(xì)胞實驗 | 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]. |
| 激酶實驗 | 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]. |
| 體外活性 | 方法：人誘導(dǎo)多能干細(xì)胞 marmoset iPSC 用 Y-27632 (5-20 μM) 處理 7 天，使用 AKP 檢測克隆形成情況。
結(jié)果：Y-27632 顯著提高 marmoset iPSC 的克隆效率。[1]
方法：成人脂肪組織衍生干細(xì)胞 ADSCs 用 Y-27632 (5 μmol/L) 處理 1 h，檢測 ADSCs 的形態(tài)變化。
結(jié)果：Y-27632 劑量依賴性誘導(dǎo) ADSCs 的神經(jīng)元分化，5 μmol/L Y-27632 處理 1 h 的 ADSCs的神經(jīng)元樣細(xì)胞百分比為（93.5±4.7）%。[2]
方法：食蟹猴胚胎干細(xì)胞 cyES 常規(guī)傳代或用 Y-27632 (1-10 μM) 處理 24 h，使用 Flow Cytometry 方法進行活-死染色，使用試劑盒檢測 BrdU。
結(jié)果：Y-27632 促進 cyES 存活細(xì)胞增加。Y-27632 沒有促進細(xì)胞增殖，但保護細(xì)胞在單細(xì)胞消化后免于細(xì)胞死亡。[3] |
| 體內(nèi)活性 | 方法：為研究 Y-27632 在運動神經(jīng)元疾病的治療潛力，將 Y-27632 (2 or 30 mg/kg in drinking water) 口服給 ALS 模型的 SOD1G93A 小鼠，持續(xù) 137 天。
結(jié)果：Y-27632 2 mg/kg 治療的效果不佳，Y-27632 30 mg/kg 治療可改善雄性小鼠的運動功能，雌性小鼠僅表現(xiàn)出有限的改善。[4]
方法：為研究 Y-27632 對肝纖維化的影響，將 Y-27632 (30 mg/kg) 口服給藥給 dimethylnitrosamine (DMN) 誘導(dǎo)肝纖維化的大鼠，每天一次，持續(xù)四周。
結(jié)果：Y-27632 治療顯著減少了 DMN 誘導(dǎo)的肝纖維化的發(fā)生，并降低了肝臟中膠原和羥脯氨酸的含量以及 α-SMA 的表達。[5] |
| 存儲條件 | keep away from direct sunlight | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice./Shipping at ambient temperature. |
| 溶解度 | H2O : insoluble
10% DMSO+90% Saline : 2.47 mg/mL (9.99 mM), Solution.
DMSO : 45 mg/mL (181.94 mM), Sonication is recommended.
|
| 關(guān)鍵字 | Y-27632 | Y27632 | Y 27632 | ROK | ROCK2 | ROCK1 | ROCK | Rho-kinase | Rho-associated protein kinase | Rho-associated kinase | Inhibitor | inhibit | Apoptosis |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Cysteamine hydrochloride | Alginic acid | Flubendazole | L-Glutamic acid monosodium salt monohydrate | 5-Fluorouracil | Stavudine | Tributyrin | Dextran sulfate sodium salt (MW 4500-5500) | L-Ascorbic acid sodium salt | L-Ascorbic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 細(xì)胞周期化合物庫 | 激酶抑制劑庫 | 神經(jīng)再生化合物庫 | 細(xì)胞凋亡化合物庫 | TGF-β/Smad靶點化合物庫 | 膜蛋白靶向化合物庫 | 口服活性化合物庫 | 表型篩選靶點鑒定庫 |