3-Methyladenine (3-MA) is a selective PI3KV inhibitor, and the IC50s against ps34 and PI3Kγ were 25/60 μM in HeLa cells, respectively.

Although 3-MA shows some limited Vps34 preference in vitro, with an IC50 of 25 μM for Vps34 as compared with 60 μM for PtdIns3Kγ it is typically employed in cellular studies at a concentration of 10 mM, which can inhibit all PtdIns3Ks [1]. The treatment of cerebellar granule cells with either 20 or 10 mM 3-MA prevented both autophagosome proliferation and cell death, without affecting neuronal morphology nor protein synthesis [2]. Treatment with 5 mM 3-MA decreased the percentage of glucose-starved HeLa cells displaying GFP-LC3 puncta to 23%. Treatment of HeLa cells with 2.5 mM or 5 mM 3-MA for one day did not affect cell viability, whereas treatment with 10 mM 3-MA for one day caused a 25.0% decrease in cell viability. Treatment of cells with 2.5, 5 or 10 mM 3-MA for two days caused 11.5%, 38.0% or 79.4% decrease in viability, respectively [3].

In severe acute pancreatitis (SAP) group, the pathological change increased with time after modeling. Pathological change of the pancreas tissue in 3-methyladenine group was milder than those in the SAP group at both 12 and 24 h [4]. 3-MA pretreatment significantly aggravated neurological symptoms when compared with the SAH + vehicle group. a large number of dying neurons from the SAH + 3-MA group showed cell shrinkage, chromatin condensation at the nuclear membrane and nuclear and cellular fragmentation, which suggest that the neurons were undergoing apoptotic cell death [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].

5142-23-4

C6H7N5

149.157

NSC 66389;3-甲基腺嘌呤;3-MA;3-Methyladenine

[1]Hou H, et al. Inhibitors of phosphatidylinositol 3'-kinases promote mitotic cell death in HeLa cells. PLoS One. 2012;7(4):e35665. [2]Miller S, et al. Finding a fitting shoe for Cinderella: searching for an autophagy inhibitor. Autophagy. 2010 Aug;6(6):805-7. [3]Zhang H, Cui Z, Cheng D, et al. RNF186 regulates EFNB1 (ephrin B1)-EPHB2-induced autophagy in the colonic epithelial cells for the maintenance of intestinal homeostasis[J]. Autophagy . 2020 [4]Shang Z, Zhang T, Jiang M, et al. High-carbohydrate, High-fat Diet-induced Hyperlipidemia Hampers the Differentiation Balance of Bone Marrow Mesenchymal Stem Cells by Suppressing Autophagy via the AMPK/mTOR Pathway in Rat Models[J]. 2020. [5]hang C, Liu Z, Zhang Y, et al. Z“Iron free” zinc oxide nanoparticles with ion-leaking properties disrupt intracellular ROS and iron homeostasis to induce ferroptosis[J]. Cell Death & Disease. 2020, 11(3): 1-15. [6]Wang S, Li F, Qiao R, et al. Arginine-Rich Manganese Silicate Nanobubbles as a Ferroptosis-Inducing Agent for Tumor-Targeted Theranostics[J]. ACS nano. 2018 Dec 26;12(12):12380-12392. [7]Xia Y, Chen J, Yu Y, et al. Compensatory combination of mTOR and TrxR inhibitors to cause oxidative stress and regression of tumors[J]. Theranostics. 2021, 11(9): 4335. [8]Canu N, et al. Role of the autophagic-lysosomal system on low potassium-induced apoptosis in cultured cerebellar granule cells. J Neurochem. 2005 Mar;92(5):1228-42. [9]Jing CH, et al. Autophagy activation is associated with neuroprotection against apoptosis via a mitochondrial pathway in a rat model of subarachnoid hemorrhage. Neuroscience. 2012 Jun 28;213:144-53. [10]Wang X, et al. Acanthopanax versus 3-Methyladenine Ameliorates Sodium Taurocholate-Induced Severe Acute Pancreatitis by Inhibiting the Autophagic Pathway in Rats. Mediators Inflamm. 2016;2016:8369704.

[1]Wang S, Li F, Qiao R, et al. Arginine-Rich Manganese Silicate Nanobubbles as a Ferroptosis-Inducing Agent for Tumor-Targeted Theranostics. ACS nano. 2018 Dec 26;12(12):12380-12392. [2]Gao X, Jiang P, Zhang Q, et al. Peglated-H1/pHGFK1 nanoparticles enhance anti-tumor effects of sorafenib by inhibition of drug-induced autophagy and stemness in renal cell carcinoma. Journal of Experimental & Clinical Cancer Researc. 2019, 38(1): 1-15 [3]Zhao Deng,Qi Liu,Miaomiao Wang,Hong-Kui Wei,Jian Peng, et al. GPA Peptide-Induced Nur77 Localization at Mitochondria Inhibits Inflammation and Oxidative Stress through Activating Autophagy in the Intestine. Oxidative Medicine and Cellular Longevity. 2020 [4]Zhao Deng,Jiangjin Ni,Xiaoyu Wu,Hongkui Wei, et al. GPA peptide inhibits NLRP3 inflammasome activation to ameliorate colitis through AMPK pathway. Aging-us. 2020 [5]Wu J N, Lin L, Luo S B, et al. SphK1‐driven autophagy potentiates focal adhesion paxillin‐mediated metastasis in colorectal cancer. Cancer Medicine. 2021 [6]Chu S, Bi H, Li X, et al. Up-regulation of Nrf2/P62/Keap1 involves in the anti-fibrotic effect of combination of monoammonium glycyrrhizinate and cysteine hydrochloride induced by CCl4. European Journal of Pharmacology. 2021: 174628. [7]Wang C, Fu J, Wang M, et al. Bartonella quintana type IV secretion effector BepE ‐induced selective autophagy by conjugation with K63 polyubiquitin chain. Cellular Microbiology. 2019, 21(4): e12984 [8]Jing Q, Li G, Chen X, et al. Wnt3a promotes radioresistance via autophagy in squamous cell carcinoma of the head and neck. Journal of Cellular and Molecular Medicine. 2019 May 21 [9]Wu A G, Teng J F, Wong V K W, et al. Novel Steroidal Saponin Isolated from Trillium tschonoskii Maxim. Exhibits Anti-Oxidative Effect via Autophagy Induction in cellular and Caenorhabditis elegans models. Phytomedicine. 2019: 153088. [10]Xiong R, Zhou X G, Tang Y, et al. Lychee seed polyphenol protects the blood–brain barrier through inhibiting Aβ (25–35)‐induced NLRP3 inflammasome activation via the AMPK/mTOR/ULK1‐mediated autophagy in bEnd. 3 cells and APP/PS1 mice. Phytotherapy Research. 2020

H2O:8 mg/mL
DMSO:3 mg/mL (20.11 mM),warmed
Ethanol:4 mg/mL (26.81 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years