| 名稱 | 3-Methyladenine |
| 描述 | 3-Methyladenine (3-MA) is a PI3K inhibitor that selectively inhibits class IB PI3Kγ (IC50=60 μM) and class III VPS34 (IC50=25 μM). 3-Methyladenine inhibits autophagy. |
| 細(xì)胞實(shí)驗(yàn) | Cells were seeded in an 8-well coverglass-bottomed chamber for 24 hours (6×10^3 cells per well). Images were acquired automatically at multiple locations on the coverglass using a Nikon TE2000E inverted microscope fitted with a 20× Nikon Plan Apo objective, a linearly-encoded stage, and a Hamamatsu Orca-ER CCD camera. A mercury-arc lamp with two neutral density filters (for a total 128-fold reduction in intensity) was used for fluorescence illumination. The microscope was controlled using NIS-Elements Advanced Research software and housed in a custom-designed 37°C chamber with a secondary internal chamber that delivered humidified 5% CO2. Fluorescence and differential interference contrast images were obtained every 10 min for a period of 48 hours. To analyze live cell imaging movies, the time-lapse records of live cell imaging experiments were exported as an image series and analyzed manually using NIS-Elements Advanced Research software. The criteria for analyses were described previously, and lagging chromosomes in prometaphase were defined as the red fluorescence-positive materials that lingered outside the roughly formed metaphase plate for more than 3 frames (30 min) [2]. |
| 動(dòng)物實(shí)驗(yàn) | All rats were fasted for 12 h with free access to water prior to operation. After anesthesia by intraperitoneal (i.p.) injection of 2% sodium pentobarbital (0.25 mL/100 g), they were laid and fixed on the table, routinely shaven, disinfected, and draped. The rat SAP model was induced by 0.1 mL/min speed uniformly retrograde infusion of a freshly prepared 3.5% sodium taurocholate solution (0.1 mL/100 g) into the biliopancreatic duct after laparotomy. Equivalent volume of normal saline solution was substituted for 3.5% sodium taurocholate solution in the sham-operation (SO) control group. The incision was closed with a continuous 3-0-silk suture, and 2 mL/100 g of saline was injected into the back subcutaneously to compensate for the fluid loss. 180 rats were randomly divided into four groups: (1) Acanthopanax treatment group (Aca group, n = 45) where the rats were injected with 0.2% Acanthopanax injection at a dose of 3.5 mg/100 g 3 h after successful modeling via the vena caudalis once, knowing that this dosage was effective as proven in our previous experiment; (2) 3-Methyladenine treatment group (3-methyladenine group, n = 45) where the rats were injected with 100 nmol/μL 3-methyladenine solution at a dose of 1.5 mg/100 g 3 h after successful modeling via the intraperitoneal route once, knowing that this dosage was effective as proven in the literature [6]; (3) SAP model group (SAP group, n = 45) where these rats received an equivalent volume of the normal saline instead of Acanthopanax injection 3 h after successful modeling via the vena caudalis once; (4) SO group (control, n = 45) where these rats received an equivalent volume of the normal saline instead of Acanthopanax injection 3 h after successful sham-operation via the vena caudalis once. The 45 animals in each of the four groups were equally randomized into 3, 12, and 24 h subgroups for postoperative observations [4]. |
| 體外活性 | 方法：人宮頸癌細(xì)胞 HeLa 用 3-Methyladenine (2.5-10 mM) 處理 48 h，使用 Trypan blue dye exclusion assay 檢測細(xì)胞生長抑制情況。
結(jié)果：3-Methyladenine 以時(shí)間和劑量依賴的方式降低 HeLa 細(xì)胞活力。[1]
方法：脂肪細(xì)胞 3T3-L1 在沒有血清的情況下用 3-Methyladenine (5 mM) 處理 4 h，使用 Western Blot 方法檢測靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：3-Methyladenine 顯著降低了自噬標(biāo)記物 LC3-II 的細(xì)胞內(nèi)水平，增加了 p62 的表達(dá)，表明 3-Methyladenine 有效抑制自噬。[2]
方法：小鼠黑色素瘤細(xì)胞 B16 用 2DG (5 mM)、rotenone (1 μM) 和 3-Methyladenine (1.2-5 mM) 處理 24 h，使用 LDH release assay 檢測細(xì)胞毒性。
結(jié)果：3-Methyladenine 劑量依賴性降低 2DG/rotenone 引起的 LDH 釋放上調(diào)，保護(hù)腫瘤細(xì)胞免受糖酵解和線粒體呼吸抑制。[3] |
| 體內(nèi)活性 | 方法：為研究 3-Methyladenine 對(duì)動(dòng)脈粥樣硬化的影響，將 3-Methyladenine (30 mg/kg) 腹腔注射給 HFD 喂養(yǎng)的 ApoE?/? 小鼠，每周兩次，持續(xù)八周。
結(jié)果：在高脂肪飲食喂養(yǎng)的小鼠中， 3-Methyladenine 治療顯著減少了動(dòng)脈粥樣硬化斑塊的大小，并增加了病變的穩(wěn)定性。3-Methyladenine 具有多種動(dòng)脈粥樣硬化保護(hù)作用，包括調(diào)節(jié)巨噬細(xì)胞自噬和泡沫細(xì)胞形成以及改變免疫微環(huán)境。[4]
方法：為研究自噬的調(diào)節(jié)作用，將 3-Methyladenine (15 mg/kg?) 單劑量腹腔注射給 LPS 誘導(dǎo)內(nèi)毒素休克的 C57/BL6 小鼠。
結(jié)果： LPS 聯(lián)合 3-Methyladenine 治療的動(dòng)物在內(nèi)毒素血癥后表現(xiàn)出存活率增加，血清炎癥介質(zhì) TNF-α 和 IL-6 降低。[5] |
| 存儲(chǔ)條件 | store at low temperature,keep away from direct sunlight,keep away from moisture | Powder: -20°C for 3 years | Shipping with blue ice. |
| 溶解度 | DMSO : 13.75 mg/mL (92.19 mM), Heating is recommended.(The compound is unstable in solution, please use soon.)
Ethanol : 4 mg/mL (26.81 mM)
H2O : 3 mg/mL (20.11 mM）, Sonication and heating are recommended. (The compound is unstable in solution, please use soon.)
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| 關(guān)鍵字 | 3 Methyladenine | inhibit | Mitochondrial Autophagy | 3Methyladenine | NSC66389 | PI3K | Endogenous Metabolite | 3-Methyladenine | Mitophagy | Autophagy | NSC-66389 | Phosphoinositide 3-kinase | Inhibitor |
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