AbMole丨A-769662：AMPK激活剂在炎症、动物肥胖中的研究应用

A-769662（AbMole，M3444）是一种特异性的AMP活化蛋白激酶（AMPK）变构激活剂，能通过结合AMPK的变构位点激活AMPK，并抑制AMPK的Thr-172位点的去磷酸化[1]。A-769662在多种细胞系中展现出显著的生物学效应：在A549人非小细胞肺癌细胞中，对细胞增殖的半数抑制浓度（IC₅₀）为45.29 ± 2.14 μM[2]；在U373胶质瘤细胞、HTAS下丘脑星形胶质细胞及CRTAS皮层星形胶质细胞中，100 μM的 A-769662可显著提升AMPK Thr172位点的磷酸化水平，并诱导胞内钙离子浓度升高，进而促进胞外ATP（eATP）释放[3]；A-769662在3T3-L1脂肪细胞中，可以20 μM的浓度有效抑制葡萄糖摄取[4]；A-769662还在骨髓间充质干细胞（mBMSCs）中，通过AMPK的依赖机制促进成骨分化并抑制脂肪生成，该效应可被AMPKα1 的siRNA或Compound C（Dorsomorphin，一种AMPK选择性抑制剂）阻断[5]；在脑微血管内皮细胞（BMEC）中，100 μM的A-769662处理16小时可增加ACC Ser79和TSC2 Ser1387的磷酸化，降低脂质合成19%[6]；在LPS（Lipopolysaccharides，脂多糖）刺激的心肌组织或原代主动脉血管内皮细胞（VECs）中，A-769662可抑制TLR-4表达及炎症反应，该作用亦可被Compound C 逆转。在动物实验方面，A-769662（AbMole，M3444）在多种啮齿类模型中被广泛应用：在C57BL/6小鼠中，腹腔注射10 mg/kg可有效激活心肌AMPK通路并减轻LPS诱导的炎症反应[7]；A-769662在糖尿病大鼠模型中，6.0 mg/kg的剂量可恢复缺血后处理（IPO）的心肌保护作用，增强自噬并减少梗死面积[8]；在高脂饮食诱导的肥胖小鼠模型中，长期低剂量给予A-769662 可改善代谢紊乱、减轻体重增加并促进棕色脂肪组织（BAT）活化及白色脂肪组织（WAT）褐变[9]；此外，在SD大鼠脑缺血模型中，A-769662（CAS No.：844499-71-4）干预可显著降低血清肌酐（Scr）、尿素氮（BUN）及γ-氨基丁酸（γ-GABA）水平，提示其对神经－肾脏轴的调节作用[10]。

参考文献及鸣谢

[1] Aledavood, E. Moraes, G. Lameira, J. et al. Understanding the Mechanism of Direct Activation of AMP-Kinase: Toward a Fine Allosteric Tuning of the Kinase Activity. Journal of chemical information and modeling 2019, 59 (6), 2859-2870.

[2] Guo, Y.; Jiang, X. Chang, Q. et al. Novel pyrazolo[3,4-b]pyridine derivatives: Synthesis, structure-activity relationship studies, and regulation of the AMPK/70S6K pathway. Archiv der Pharmazie 2022, 355 (7), e2100465.

[3] Vlachaki Walker, J. M. Robb, J. L. Cruz, A. M. et al. AMP-activated protein kinase (AMPK) activator A-769662 increases intracellular calcium and ATP release from astrocytes in an AMPK-independent manner. Diabetes, obesity & metabolism 2017, 19 (7), 997-1005.

[4] Kopietz, F. Alshuweishi, Y.; Bijland, S. et al. A-769662 inhibits adipocyte glucose uptake in an AMPK-independent manner. The Biochemical journal 2021, 478 (3), 633-646.

[5] Abdallah, B. M. Alzahrani, A. M. A-769662 stimulates the differentiation of bone marrow-derived mesenchymal stem cells into osteoblasts via AMP-activated protein kinase-dependent mechanism. Journal of applied biomedicine 2021, 19 (3), 159-169.

[6] Huang, J. Guesthier, M. A. Burgos, S. A. AMP-activated protein kinase controls lipid and lactose synthesis in bovine mammary epithelial cells. Journal of dairy science 2020, 103 (1), 340-351.

[7] Rameshrad, M. Maleki-Dizaji, N. Soraya, H. et al. Effect of A-769662, a direct AMPK activator, on Tlr-4 expression and activity in mice heart tissue. Iranian journal of basic medical sciences 2016, 19 (12), 1308-1317.

[8] Zhou, B. Leng, Y.; Lei, S. Q. et al. AMPK activation restores ischemic post‑conditioning cardioprotection in STZ‑induced type 1 diabetic rats: Role of autophagy. Molecular medicine reports 2017, 16 (3), 3648-3656.

[9] Desjardins, E. M. Steinberg, G. R. Emerging Role of AMPK in Brown and Beige Adipose Tissue (BAT): Implications for Obesity, Insulin Resistance, and Type 2 Diabetes. Current diabetes reports 2018, 18 (10), 80.

[10] Yang, L. Gong, N. R. Zhang, Q. et al. Apparent Correlations Between AMPK Expression and Brain Inflammatory Response and Neurological Function Factors in Rats with Chronic Renal Failure. Journal of molecular neuroscience : MN 2019, 68 (2), 204-213.