Real Time Monitoring of Inhibition of Adipogenesis and Angiogenesis by (-)-Epigallocatechin-3-Gallate in 3T3-L1 Adipocytes and Human Umbilical Vein Endothelial Cells.
Real Time Monitoring of Inhibition of Adipogenesis and Angiogenesis by (-)-Epigallocatechin-3-Gallate in 3T3-L1 Adipocytes and Human Umbilical Vein Endothelial Cells.
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DOI：
10.3390/nu7105437
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发表时间：
2015-10-27
期刊：
影响因子：
5.9
通讯作者：
Shen X
中科院分区：
文献类型：
--
作者：
Tang W;Song H;Cai W;Shen X
表没食子儿茶素没食子酸酯(−)对脂肪细胞血管生成的影响目前知之甚少。本研究旨在检测EGCG对脂肪细胞血管内皮生长因子(VEGF)表达的影响。采用酶联免疫吸附试验、实时定量聚合酶链式反应和免疫荧光染色分别检测3T3-L1细胞的血管内皮生长因子分泌水平、血管内皮生长因子信使核糖核酸表达水平和血管内皮生长因子蛋白表达水平。采用xCELLigence实时细胞分析系统对3T3-L1前脂肪细胞的生长分化进行研究。采用共培养系统检测3T3-L1细胞对人脐静脉内皮细胞(HUVECs)增殖的影响。用加或不加EGCG的3T3-L1细胞的条件培养液培养人脐静脉内皮细胞，进行成管实验。通过检测与脂肪生成和血管生成相关的两个转录因子--过氧化物酶体增殖物激活受体γ(PPARγ)和CCAAT/增强子结合蛋白α(C/EBPα)，探讨其可能的机制。我们发现，经EGCG处理后，脂肪细胞中血管内皮生长因子表达的三项指标(mRNA、蛋白和分泌物)均降低。与3T3-L1细胞共培养的HUVECs生长明显增加，EGCG处理的3T3-L1脂肪细胞的条件培养液抑制了HUVECs的管状形成。EGCG可降低脂肪细胞PPARγ和C/EBPα的表达。综上所述，本研究结果提示，EGCG可能通过调节脂肪细胞中血管内皮生长因子的表达和分泌来抑制血管生成。
Little is known about the effect of (−)-epigallocatechin-3-gallate (EGCG) on angiogenesis in adipocytes. We aimed to test the effect of EGCG on the expression of vascular endothelial growth factor (VEGF) in adipocytes. The levels of VEGF secretion, the expression of VEGF message ribonucleic acid (mRNA) and VEGF protein in 3T3-L1 cells were measured by enzyme linked immunosorbent assay (ELISA), real time polymerase chain reaction (PCR), and immunofluorescence staining, respectively. The xCELLigence real time cell analysis system was used to study the growth and differentiation of 3T3-L1 preadipocytes. A coculture system was used to test the effects of 3T3-L1 cells on proliferation of human umbilical vein endothelial cells (HUVECs). The conditioned media derived from 3T3-L1 cells treated with or without EGCG was used to culture the HUVECs for a tube formation assay. Peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα), two transcription factors related to both adipogenesis and angiogenesis, were examined to explore the potential mechanism. We found that all the three measurements of VEGF expression in adipocytes (mRNA, protein and secretion in media) were reduced after EGCG treatment. The growth of HUVECs co-cultured with 3T3-L1 cells was significantly increased and the conditioned media from EGCG treated 3T3-L1 adipocytes inhibited tube formation in HUVECs. Both PPARγ and C/EBPα expression in adipocytes were decreased with EGCG treatment. In conclusion, findings from this study suggest that EGCG may inhibit angiogenesis by regulating VEGF expression and secretion in adipocytes.