| 名稱(chēng) | Forskolin |
| 描述 | Forskolin (Coleonol) is a natural product, an adenylate cyclase activator (EC50=0.5 μM). Forskolin increases cAMP levels, activates PXR and FXR, and induces autophagy. Forskolin produces positive inotropic effects in the heart, and has platelet anticoagulant and antihypertensive effects. |
| 細(xì)胞實(shí)驗(yàn) | Kit 225 or MT-2 cells were treated with 1, 5, 10, 20, 50, or 100 μM Forskolin for 20 min at 37 °C. Cells were lysed and clarified by centrifugation, and the concentration of cAMP was detected by direct cAMP ELISA. Optical density was measured at 405 nm, and the concentration of intracellular cAMP was calculated using a weighted four parameter logistic curve according to the manufactures instructions [2]. |
| 激酶實(shí)驗(yàn) | For Jak3 kinase assays, Fsk-treated MT-2 cells were lysed, clarified, and immunoprecipitated using Jak3 antibody as described above. Kinase reactions were carried out as described previously at 30 °C for 20 min. For PKA kinase assays, untreated MT-2 cells were lysed, and Jak3 was immunoprecipitated and bound to PAS beads as described previously. Immunoprecipitated Jak3 was washed with kinase buffer (50 mM Hepes-NaOH (pH 7.4), 10 mM MgCl2, 0.5 mM EGTA, 0.5 mM DTT, 20 μg/ml aprotinin, 10 μg/ml leupeptin, 1 μg/ml pepstatin A) and incubated with 200 μM ATP and purified protein kinase A catalytic subunit (PKAc) as indicated in the figure legends. Kinase reactions were carried out at 32 °C for 30 min followed by vigorous washing of the beads with cold kinase wash buffer as described previously. For [γ-32P]ATP radiolabeled kinase assays using recombinant Jak3, Hek293 cells were transfected with wild type (WT) Jak3 or kinase-dead Jak3 K855A using Lipofectamine 2000 according to the manufacturer's instructions. Cells were lysed and immunoprecipitated with Jak3 antibody. Jak3-bound PAS beads were washed three times in cold lysis buffer followed by kinase buffer. Kinase reactions were initiated by adding 10 μCi [γ-32P]ATP, 10 μm unlabeled ATP, and 1 μg of purified PKAc to Jak3-bound PAS bead reaction mixtures. Kinase reactions were performed at 32 °C for 30 min. Jak3-bound PAS beads were washed three times in radioimmunoassay buffer (10 mM Tris-HCl, pH 7.4, 75 mM NaCl, 20 mM EDTA, 10 mM EGTA, 20 mM Na4P2O7, 50 mM NaF, 20 mM 2-glycerolphosphate, 1 mM p-nitrophenyl phosphate, 0.1% Triton X-100) and one time in kinase wash buffer. The reactions were stopped by adding 2× SDS-PAGE sample buffer followed by SDS-PAGE. Coomassie stainable Jak3 bands were excised from the PVDF membrane and subjected to phosphoamino acid analysis [2]. |
| 動(dòng)物實(shí)驗(yàn) | Forskolin was dissolved in dimethyl sulfoxide (DMSO) and injected intraperitoneally into neonatal mice at postnatal days 4 (P4) and 5 (P5). Mice injected with DMSO served as the controls. The treated mice were euthanized at P6, and their retinas were isolated for whole-mount immunohistochemistry (IHC). We first tested the effect of different concentrations of forskolin on the survival rate and retinal vasculature and determined the optimal concentration, 1.0 μg/50 μL (0.3 mg/kg) at P4 and 1.5 μg/50 μL (0.5 mg/kg) at P5, used to compare the retinal vascular phenotypes between WT mice and Mrp4-deficient mice [4]. . After acclimatization for 2 weeks, animals were randomly divided into four groups of eight rats each and treated for six consecutive weeks as follows: The first group was treated with CCl4 (50% CCl4/corn oil; 0.5 mL·kg?1, i.p.) twice a week to induce liver fibrosis. The second group was given forskolin only at a dose of 10 mg·kg?1, i.p., dissolved in a DMSO/saline solution (1:49) five times a week. The third group was given both CCl4 and forskolin. The dose of forskolin used here was based on the results of our preliminary study. The fourth group served as the normal control, receiving vehicles only. At 24 h after the last injection, blood samples were collected from the retro‐orbital plexus after light anesthesia with sodium pentobarbital (50 mg·kg?1, i.p.). Serum was separated by centrifugation at 3000× g for 10 min and was used for the assessment of liver functions. Rats were killed by cervical dislocation, and livers were removed and weighed. A portion of liver tissue was washed and homogenized to obtain a 20% (w·v?1) homogenate, which was used for assessment of oxidative stress, inflammatory and fibrogenic markers. Another portion was placed in formalin for immunohistochemical and histopathological analyses. The remainder was stored at ?80°C, together with the 20% homogenate, until needed [5]. |
| 體外活性 | 方法：大鼠腎上腺髓質(zhì)嗜鉻瘤細(xì)胞 PC12 用 Forskolin (0.01-10 μM) 處理 3-48 h，使用 MTT 方法檢測(cè)細(xì)胞生長(zhǎng)抑制情況。
結(jié)果：用 10μM Forskolin 處理后，細(xì)胞活力迅速下降，處理 6 h 后細(xì)胞活力下降為 88.4%，處理 48 h 后細(xì)胞活力下降為 60.5%。[1]
方法：人骨髓瘤細(xì)胞 U266、H929、INA-6、RPMI 8226 和 OPM-2 用 Forskolin (1-100 μM) 處理 72 h，使用 Flow Cytometry 方法檢測(cè)細(xì)胞死亡情況。
結(jié)果：Forskolin 劑量依賴(lài)性誘導(dǎo)人骨髓瘤細(xì)胞死亡，其中 U266、OPM-2 和 INA-6 比 H929 和 RPMI 8226 細(xì)胞更敏感。[2]
方法：人 IL-2 依賴(lài)性白血病細(xì)胞 Kit 225 和人白血病細(xì)胞 MT-2 用 Forskolin (1-100 μM) 處理 20 min，使用 ELISA 方法測(cè)定 cAMP 濃度。
結(jié)果：Forskolin 誘導(dǎo) cAMP 水平上調(diào)，在 50-100 μm 之間達(dá)到最大水平。[3] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Forskolin (4-5 mg/kg in PBS/DMSO solution (15:1)) 腹腔注射給攜帶鼠多發(fā)性骨髓瘤腫瘤 MOPC315 的 BALB/c nude 小鼠，在腫瘤細(xì)胞注射后的第 2/4/6 天給藥。
結(jié)果：所有小鼠最終都發(fā)生了腫瘤，但 Forskolin 顯著延緩了體內(nèi)腫瘤的生長(zhǎng)。提高 cAMP 的化合物可能在治療多發(fā)性骨髓瘤方面具有治療潛力。[4]
方法：為研究 Forskolin 對(duì)糖尿病條件下視網(wǎng)膜炎癥的影響，將 Forskolin (50 mg/kg) 灌胃給藥給 STZ 誘導(dǎo)糖尿病模型的 C57BL/6 小鼠，每周一次，持續(xù)十二周。
結(jié)果：與正常對(duì)照組相比，糖尿病對(duì)照組和 Forskolin 治療組的視網(wǎng)膜葡萄糖濃度均增加，但由于葡萄糖轉(zhuǎn)運(yùn)蛋白 1 表達(dá)下調(diào)，F(xiàn)orskolin 處理組僅為糖尿病對(duì)照組的約 68.06%。與正常對(duì)照組相比，F(xiàn)orskolin 治療組和糖尿病對(duì)照組的 ICAM?1 和 TNF-α 表達(dá)上調(diào)，但 Forskolin 處理組的這兩種炎癥因子表達(dá)水平分別為糖尿病對(duì)照的 68.75% 和 75.37%。[5] |
| 存儲(chǔ)條件 | keep away from direct sunlight,keep away from moisture,store at low temperature | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : Insoluble
DMSO : 55 mg/mL (133.98 mM)
Ethanol : 15 mg/mL (36.5 mM)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 3 mg/mL (7.31 mM), Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
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| 關(guān)鍵字 | notropic | pregnane | Inhibitor | exosome | cancer | antihypertensive | Adenylyl cyclase | antiaggregatory | X receptor | prostate | Forskolin | NR1H4 | Adenylate Cyclase | PXR | cAMP | inhibit | FXR | Autophagy |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | Taurine | Gefitinib | Aceglutamide | Hydroxychloroquine | Curcumin | Stavudine | Salicylic acid | Paeonol | Sodium 4-phenylbutyrate |
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