A-769662作為AMPK激活劑在炎癥、動(dòng)物肥胖和腫瘤模型中的研究應(yīng)用

A-769662（AbMole，M3444）是一種特異性的AMP活化蛋白激酶（AMPK）變構(gòu)激活劑，能通過結(jié)合AMPK的變構(gòu)位點(diǎn)激活A(yù)MPK，并抑制AMPK的Thr-172位點(diǎn)的去磷酸化[1]。A-769662在多種細(xì)胞系中展現(xiàn)出顯著的生物學(xué)效應(yīng)：在A549人非小細(xì)胞肺癌細(xì)胞中，對細(xì)胞增殖的半數(shù)抑制濃度（IC₅₀）為45.29 ± 2.14 μM[2]；在U373膠質(zhì)瘤細(xì)胞、HTAS下丘腦星形膠質(zhì)細(xì)胞及CRTAS皮層星形膠質(zhì)細(xì)胞中，100 μM的 A-769662可顯著提升AMPK Thr172位點(diǎn)的磷酸化水平，并誘導(dǎo)胞內(nèi)鈣離子濃度升高，進(jìn)而促進(jìn)胞外ATP（eATP）釋放[3]；A-769662在3T3-L1脂肪細(xì)胞中，可以20 μM的濃度有效抑制葡萄糖攝取[4]；A-769662還在骨髓間充質(zhì)干細(xì)胞（mBMSCs）中，通過AMPK的依賴機(jī)制促進(jìn)成骨分化并抑制脂肪生成，該效應(yīng)可被AMPKα1 的siRNA或Compound C（Dorsomorphin，一種AMPK選擇性抑制劑）阻斷[5]；在腦微血管內(nèi)皮細(xì)胞（BMEC）中，100 μM的A-769662處理16小時(shí)可增加ACC Ser79和TSC2 Ser1387的磷酸化，降低脂質(zhì)合成19%[6]；在LPS（Lipopolysaccharides，脂多糖）刺激的心肌組織或原代主動(dòng)脈血管內(nèi)皮細(xì)胞（VECs）中，A-769662可抑制TLR-4表達(dá)及炎癥反應(yīng)，該作用亦可被Compound C 逆轉(zhuǎn)。在動(dòng)物實(shí)驗(yàn)方面，A-769662（AbMole，M3444）在多種嚙齒類模型中被廣泛應(yīng)用：在C57BL/6小鼠中，腹腔注射10 mg/kg可有效激活心肌AMPK通路并減輕LPS誘導(dǎo)的炎癥反應(yīng)[7]；A-769662在糖尿病大鼠模型中，6.0 mg/kg的劑量可恢復(fù)缺血后處理（IPO）的心肌保護(hù)作用，增強(qiáng)自噬并減少梗死面積[8]；在高脂飲食誘導(dǎo)的肥胖小鼠模型中，長期低劑量給予A-769662 可改善代謝紊亂、減輕體重增加并促進(jìn)棕色脂肪組織（BAT）活化及白色脂肪組織（WAT）褐變[9]；此外，在SD大鼠腦缺血模型中，A-769662（CAS No.：844499-71-4）干預(yù)可顯著降低血清肌酐（Scr）、尿素氮（BUN）及γ-氨基丁酸（γ-GABA）水平，提示其對神經(jīng)－腎臟軸的調(diào)節(jié)作用[10]。
 
參考文獻(xiàn)及鳴謝
[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.