南方医科大学学报2017，Vol.37Issue(5)：614-621,8.DOI:10.3969/j.issn.1673-4254.2017.05.08
球状脂联素对卵巢微血管内皮细胞增殖、迁移及管状结构形成的影响
Effect of globular adiponectin on proliferation, migration and tube formation of ovarian microvascular endothelial cells
摘要
Abstract
Objective To analyze the effect of globular adiponectin on angiogenesis of ovarian microvascular endothelial cells (OMECs).Methods Mouse OMECs were isolated and purified by density gradient centrifugation with Percoll and identified by immunofluorescence analysis of follicle-stimulating hormone receptor (FSHR),luteinizing hormone receptor (LHR),and endothelial cell marker von Willebrand factor (vWF).The capillary-like tube formation of OMECs was determined by vascular endothelial growth factor A (VEGFA) treatment in Matrigel matrix.OMECs treated with recombinant globular adiponectin protein were examined for cell proliferation with MTS assay and cell migration with scratch wound healing assay,and capillary-like tube formation was tested in Matrigel matrix.Western blotting was performed to detect the effect of globular adiponectin on AMPK phosphorylation.Results The signals of LHR and vWF,but not that of FSHR,were detected in the isolated cells.VEGFA treatment of the cells induced capillary-like tube formation,indicating their properties of ovarian-specific endothelial cells.Treatment with 1 and 3 μg/mL of recombinant globular adiponectin significantly increased the number of OMECs by (158.72±14.50) ％ and (186.50±4.20)％ (P＜0.01) and resulted in scratch wound closure rates of (49.43±3.43)％ (P＜0.05) and (69.67±1.2) ％ (P＜0.01) respectively.The cells treated with 3 μg/mL globular adiponectin formed a capillary-tube length 6.63± 0.66 folds greater than that formed by the control cells (P＜0.01).Treatment of the cells with 3 μg/mL globular adiponectin for 15 and 30 min resulted in pAMPK/AMPK ratios of 0.86 ± 0.08 and 0.66 ± 0.13,respectively significantly higher than that in the control cells (0.13± 0.12,P＜0.01).Compound C obviously suppressed the tube formation and AMPK phosphorylafion induced by globular adiponectin.Conclusion Globular adiponectin promotes angiogenesis of OMECs through activation of the AMPK signal pathway.关键词
球状脂联素/卵巢/血管新生/血管内皮细胞/AMPKKey words
globular adiponectin/ovary/angiogenesis/vascular endothelial cells/AMPK引用本文复制引用
陈雷,卢小圣,李雅兰,毛周飞,肖銮娟,禹艳红..球状脂联素对卵巢微血管内皮细胞增殖、迁移及管状结构形成的影响[J].南方医科大学学报,2017,37(5):614-621,8.基金项目
国家自然科学基金(30900232) （30900232）
中央高校基本科研业务费专项资金(21610202) （21610202）
天河区科技计划项目(201404KW041) （201404KW041）
广东省优秀博士学位论文作者资助项目(sybzzxm201033) Supported by National Natural Science Foundation of China (30900232). （sybzzxm201033）
