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.

Adenylyl cyclase:0.5 μM (cell free)

方法：大鼠肾上腺髓质嗜铬瘤细胞 PC12 用 Forskolin (0.01-10 µM) 处理 3-48 h，使用 MTT 方法检测细胞生长抑制情况。
结果：用 10μM Forskolin 处理后，细胞活力迅速下降，处理 6 h 后细胞活力下降为 88.4%，处理 48 h 后细胞活力下降为 60.5%。[1]
方法：人骨髓瘤细胞 U266、H929、INA-6、RPMI 8226 和 OPM-2 用 Forskolin (1-100 µM) 处理 72 h，使用 Flow Cytometry 方法检测细胞死亡情况。
结果：Forskolin 剂量依赖性诱导人骨髓瘤细胞死亡，其中 U266、OPM-2 和 INA-6 比 H929 和 RPMI 8226 细胞更敏感。[2]
方法：人 IL-2 依赖性白血病细胞 Kit 225 和人白血病细胞 MT-2 用 Forskolin (1-100 μM) 处理 20 min，使用 ELISA 方法测定 cAMP 浓度。
结果：Forskolin 诱导 cAMP 水平上调，在 50-100 μm 之间达到最大水平。[3]

方法：为检测体内抗肿瘤活性，将 Forskolin (4-5 mg/kg in PBS/DMSO solution (15:1)) 腹腔注射给携带鼠多发性骨髓瘤肿瘤 MOPC315 的 BALB/c nude 小鼠，在肿瘤细胞注射后的第 2/4/6 天给药。
结果：所有小鼠最终都发生了肿瘤，但 Forskolin 显著延缓了体内肿瘤的生长。提高 cAMP 的化合物可能在治疗多发性骨髓瘤方面具有治疗潜力。[4]
方法：为研究 Forskolin 对糖尿病条件下视网膜炎症的影响，将 Forskolin (50 mg/kg) 灌胃给药给 STZ 诱导糖尿病模型的 C57BL/6 小鼠，每周一次，持续十二周。
结果：与正常对照组相比，糖尿病对照组和 Forskolin 治疗组的视网膜葡萄糖浓度均增加，但由于葡萄糖转运蛋白 1 表达下调，Forskolin 处理组仅为糖尿病对照组的约 68.06%。与正常对照组相比，Forskolin 治疗组和糖尿病对照组的 ICAM‑1 和 TNF-α 表达上调，但 Forskolin 处理组的这两种炎症因子表达水平分别为糖尿病对照的 68.75% 和 75.37%。[5]

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].

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].

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].