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Abstract：
GDM, as a metabolic disease during pregnancy, regulates GLUT3 translocation by AMPK, thereby affecting glucose uptake in trophoblasts. It provides a new research idea and therapeutic target for alleviating intrauterine hyperglycemia in GDM. STZ was used to construct GDM mice, inject AICAR into pregnant mice, and observe fetal and placental weight; flow cytometry was employed for the detection of glucose uptake by primary trophoblast cells; immunofluorescence was applied to detect the localization of GLUT3 and AMPK in placental tissue; Cocofal microscope was used to detect the localization of GLUT3 in trophoblast cells;qRT-PCR and Western blot experiments were carried out to detect the expression levels of GLUT3 and AMPK in placental tissue; CO-IP was utilized to detect the interaction of GLUT3 and AMPK. Compared with the normal pregnancy group, the weight of the fetus and placenta of GDM mice increased (P < 0.001), and the ability of trophoblasts to take up glucose decreased (P < 0.001). In addition, AMPK activity in trophoblasts and membrane localization of GLUT3 in GDM mice were down-regulated compared with normal pregnant mice (P < 0.05). There is an interaction between GLUT3 and AMPK. Activating AMPK in trophoblasts can up-regulate the expression of GLUT3 membrane protein in trophoblasts of mice (P < 0.05) and increase the glucose uptake of trophoblasts (P < 0.05). We speculate that inhibition of AMPK activity in GDM mice results in aberrant localization of GLUT3, which in turn attenuates glucose uptake by placental trophoblast cells. AICAR activates AMPK to increase the membrane localization of GLUT3 and improve the glucose uptake capacity of trophoblasts.
摘要：
GDM,作为怀孕期间的代谢疾病，通过AMPK调节GLUT3易位，从而影响滋养细胞的葡萄糖摄取。为缓解GDM宫内高血糖提供了新的研究思路和治疗靶点。STZ用于构建GDM小鼠，将AICAR注射到怀孕的小鼠体内，并观察胎儿和胎盘重量；流式细胞术用于检测原代滋养细胞对葡萄糖的摄取；免疫荧光用于检测GLUT3和AMPK在胎盘组织中的定位；Cocofal显微镜用于检测GLUT3在滋养细胞中的定位；进行qRT-PCR和Westernblot实验以检测GLUT3和AMPK在胎盘组织中的表达水平；并利用CO-IP检测GLUTK的相互作用。与正常妊娠组相比,GDM小鼠的胎儿和胎盘重量增加（P<0.001），滋养细胞吸收葡萄糖的能力下降(P<0.001)。此外,与正常妊娠小鼠相比，GDM小鼠滋养细胞AMPK活性和GLUT3的膜定位下调（P<0.05）。GLUT3和AMPK之间存在相互作用。激活滋养层AMPK可上调小鼠滋养层GLUT3膜蛋白的表达(P<0.05),增加滋养层葡萄糖的摄取(P<0.05)。我们推测，GDM小鼠中AMPK活性的抑制导致GLUT3的异常定位，进而减弱胎盘滋养层细胞对葡萄糖的摄取。AICAR激活AMPK以增加GLUT3的膜定位并提高滋养细胞的葡萄糖摄取能力。
