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.

VPS34:25 μM (in HeLa cells), PI3Kγ:60 μM (in HeLa cells)

方法：人宫颈癌细胞 HeLa 用 3-Methyladenine (2.5-10 mM) 处理 48 h，使用 Trypan blue dye exclusion assay 检测细胞生长抑制情况。
结果：3-Methyladenine 以时间和剂量依赖的方式降低 HeLa 细胞活力。[1]
方法：脂肪细胞 3T3-L1 在没有血清的情况下用 3-Methyladenine (5 mM) 处理 4 h，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：3-Methyladenine 显著降低了自噬标记物 LC3-II 的细胞内水平，增加了 p62 的表达，表明 3-Methyladenine 有效抑制自噬。[2]
方法：小鼠黑色素瘤细胞 B16 用 2DG (5 mM)、rotenone (1 μM) 和 3-Methyladenine (1.2-5 mM) 处理 24 h，使用 LDH release assay 检测细胞毒性。
结果：3-Methyladenine 剂量依赖性降低 2DG/rotenone 引起的 LDH 释放上调，保护肿瘤细胞免受糖酵解和线粒体呼吸抑制。[3]

方法：为研究 3-Methyladenine 对动脉粥样硬化的影响，将 3-Methyladenine (30 mg/kg) 腹腔注射给 HFD 喂养的 ApoE−/− 小鼠，每周两次，持续八周。
结果：在高脂肪饮食喂养的小鼠中， 3-Methyladenine 治疗显著减少了动脉粥样硬化斑块的大小，并增加了病变的稳定性。3-Methyladenine 具有多种动脉粥样硬化保护作用，包括调节巨噬细胞自噬和泡沫细胞形成以及改变免疫微环境。[4]
方法：为研究自噬的调节作用，将 3-Methyladenine (15 mg/kg ) 单剂量腹腔注射给 LPS 诱导内毒素休克的 C57/BL6 小鼠。
结果： LPS 联合 3-Methyladenine 治疗的动物在内毒素血症后表现出存活率增加，血清炎症介质 TNF-α 和 IL-6 降低。[5]

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].

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].