| 名稱(chēng) | Y-27632 dihydrochloride |
| 描述 | Y-27632 dihydrochloride (Y-27632 2HCl) is an orally potent, ATP-competitive inhibitor of ROCK-I and ROCK-II. Y-27632 dihydrochloride also inhibits isolation-induced apoptosis in mouse prostate stem or progenitor cells. |
| 細(xì)胞實(shí)驗(yàn) | HeLa cells are plated at a density of 3×10^4 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) or vehicle is added and the cells are incubated for another 30 min [1]. |
| 激酶實(shí)驗(yàn) | Recombinant ROCK1/2, 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 phosphatidylserine, 50 ng/mL 12-O-tetradecanoyl phorbol-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]. |
| 動(dòng)物實(shí)驗(yàn) | A group of animals was injected with a single dose of pentylenetetrazole (PTZ, 65?mg/kg) to investigate if the two Rho-kinase inhibitors, fasudil, and Y-27632, changed the onset of PTZ seizures. Fasudil, Y-27632 or saline was given intraperitoneally 30?min before the PTZ injection. Each mouse was then observed for a 15-min period to measure the onset of the first myoclonic jerk, the onset of the first clonic convulsion and the occurrence of tonic hindlimb extension. Some of the animals died after tonic hindlimb extension, which is an expected outcome of acute PTZ injection. After the observation period, all animals were killed by halothane anesthesia [5]. Seven-week-old male Wistar rats were anesthetized with sodium pentobarbital. A silver clip (0.2 mm in diameter) was placed on the left renal artery in the preparation of the renal hypertensive rats. In the preparation of the DOCA-salt hypertensive rats, the left kidney was removed and a DOCA pellet (50 mg) was implanted subcutaneously. The DOCA rats were then fed an 8% salt diet. Rats from both groups were used after 8 weeks in the experiments, together with a male, 17–22-week old spontaneously hypertensive rats. The average systolic pressure in these groups of hypertensive rats ranged from 209 to 237 mm Hg, and no significant difference was found between groups. Eight-week-old male Wistar rats were used as controls. Their average systolic pressure was 139 mm Hg. Y-27632was administered orally. The systolic blood pressure was measured by the tail cuff method at 1, 3, 5, 7 and 24 h. The rats were prewarmed to 40 8C for 10 min before each measurement. No toxicity was found in rats treated with 30 mg kg?1 of Y-27632 administered per os once per day for 10 days [4]. |
| 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(cè)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(cè) ADSCs 的形態(tài)變化。
結(jié)果：Y-27632 劑量依賴(lài)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(jìn)行活-死染色，使用試劑盒檢測(cè) BrdU。
結(jié)果：Y-27632 促進(jìn) cyES 存活細(xì)胞增加。Y-27632 沒(méi)有促進(jìn)細(xì)胞增殖，但保護(hù)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(yùn)動(dòng)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(yùn)動(dòng)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(duì)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(dá)。[5] |
| 存儲(chǔ)條件 | keep away from moisture,keep away from direct sunlight,store at low temperature | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 50 mg/mL (156.12 mM)
H2O : 32.03 mg/mL (100 mM), Sonication is recommended.
|
| 關(guān)鍵字 | pluripotent | orally | Y 27632 Dihydrochloride | Y-27632 Dihydrochloride | active | inhibit | Apoptosis | epithelial-mesenchymal | Y27632 Dihydrochloride | Y27632 dihydrochloride | Rho-kinase | ROK | ATP-competitive | Rho-associated kinase | modulation | lineage | ROCK | transition-like | stem | cells | Inhibitor | Y 27632 | mesendodermal | Rho-associated protein kinase | hIPSCs | Y-27632 | Y27632 | Y 27632 dihydrochloride |
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