【摘要】 目的:基于单磷酸腺苷活化蛋白激酶/环氧合酶-2(AMPK/COX-2)通路探究银杏双黄酮（GBB）对黑色素瘤细胞发生凋亡的影响。方法：体外培养人黑色素瘤细胞株A875，用不同浓度（50μmol/L、100μmol/L、200μmol/L）GBB、10μmol/L AMPK抑制剂Compound C、200μmol/L GBB+10μmol/L Compound C作用于A875细胞24 h。采用倒置显微镜观察细胞形态，Annexin V-FITC/PI双染色法检测GBB对A875细胞凋亡的影响，Western blotting法检测磷酸化乙酰辅酶A羧化酶（p-ACC）、磷酸化AMPK(p-AMPK)及COX-2蛋白表达。结果：随着GBB浓度的增加，细胞凋亡率升高（P<0.05），细胞形态回缩变小、变圆、变亮，细胞间接触消失，部分细胞脱落裂解为碎片状，悬浮细胞增多，折光性减弱，呈现典型的细胞凋亡形态。GBB可提高pACC、p-AMPK蛋白表达，降低COX-2蛋白表达（均P<0.05）;Compound C逆转了GBB对p-ACC、p-AMPK蛋白表达的促进作用及对COX-2蛋白表达的抑制作用（均P<0.05）。结论：GBB可诱导人黑色素瘤A875细胞凋亡，且呈剂量依赖性，此过程可能是通过介导AMPK/COX-2通路实现的。更多还原

【Abstract】 Objective:To investigate the effect of ginkgo biloba biflavones(GBB) on the apoptosis of melanoma cells based on adenosine 5’-monophosphate activated protein kinase/cyclooxygenase-2(AMPK/COX-2) pathway.Methods: Human melanoma cell line A875 cells were cultured in vitro, and treated with 0 μmol/L, 50 μmol/L,100 μmol/L, 200 μmol/L GBB, 10 μmol/L Compound C(AMPK inhibitor), or 200 μmol/L GBB + 10 μmol/L Compound C respectively for 24 h. An inverted microscope was used to observe the cell morphology. Annexin VFITC/PI double staining was used to detect the effect of GBB on apoptosis of A875 cells. Western blotting was used to detect the expressions of phosphorylated(p-) acetyl-CoA carboxylase(ACC), p-AMPK and COX-2 protein. Results: With the increase of GBB concentration, the apoptosis rate of A875 cells increased(P<0.05). The cell morphology shrank, became smaller, rounder and brighter, and the contact between cells disappeared. Some cells fell off and lysed into fragments, the suspended cells increased, and the refraction also weakened, showing a typical phenomenon of apoptosis. GBB could induce apoptosis of A875 cells, and with the increase of GBB concentration, the rate of apoptosis also increased(P<0.05). GBB increased the expression of p-ACC and p-AMPK while reduced the expression of COX-2 protein(P<0.05). Compound C reversedthe promotive effect of GBB on the expressions of p-ACC and p-AMPK and inhibitive effect on COX-2 protein(P<0.05). Conclusion: GBB can induce apoptosis of human melano-ma A875 cells in a dose-dependent manner, which may be achieved by mediating AMPK/COX-2 pathway.更多还原