HCG对人子宫内膜细胞VEGF表达调控的初步研究

发布时间：2018-06-30 17:34

本文选题：人子宫内膜细胞 + HCG　；参考：《现代妇产科进展》2015年12期

【摘要】：目的:探讨人子宫内膜细胞体外培养模型中,HCG对VEGF表达的调控作用和机制。方法:采用机械分离加梯度离心法分别获得腺上皮与基质细胞进行体外原代细胞混合培养;实验设对照组、HCG组(对照组使用常规DF/12培养液提取总蛋白;HCG组在DF/12培养液中加终浓度为20U/ml HCG)。采用Q-PCR测定子宫内膜细胞中VEGF mRNA表达水平。免疫组化或荧光法进行LH-R和VEGF蛋白质定位表达,Western blot法测定蛋白半定量表达。结果:子宫内膜腺上皮与基质细胞均存在LH/HCG-R和VEGF蛋白表达。HCG对子宫内膜细胞VEGF蛋白的峰值作用时间点为24h。与对照组相比,20IU/ml HCG体外干预24h后,子宫内膜细胞VEGF mRNA和蛋白质水平均显著上调(Q-PCR:1 vs 2.0132±0.6024;WB:0.4069±0.1897 vs 0.7781±0.1651)(P均0.05)。结论:成功建立了人子宫内膜细的体外培养模型,并完成HCG的干预实验。HCG与HCG/LH-R结合后上调子宫内膜细胞VEGF的基因和蛋白质水平表达。
[Abstract]:Aim: to investigate the regulation and mechanism of HCG on VEGF expression in human endometrial cells in vitro. Methods: the glandular epithelium and stromal cells were obtained by mechanical separation and gradient centrifugation respectively. The control group was established as HCG group (the control group used the conventional DF- / 12 medium to extract total protein HCG and the final concentration of HCG was 20U / ml HCG in DF- / 12 medium). The expression of VEGF mRNA in endometrial cells was measured by Q-PCR. The expression of LH-R and VEGF protein was detected by immunohistochemistry or fluorescence method. Western blot method was used to detect the semi-quantitative expression of LH-R and VEGF protein. Results: the expression of LH / HCG-R and VEGF protein in endometrial glandular epithelium and stromal cells. Compared with the control group, the mRNA and protein levels of VEGF in endometrial cells were significantly up-regulated (Q-PCR: 1 vs 2.0132 卤0.6024 WB: 0.4069 卤0.1897 vs 0.7781 卤0.1651) after intervention with 20IUU / ml HCG for 24 hours in vitro. Conclusion: the model of human endometrial fine culture in vitro was successfully established, and the HCG intervention experiment. HCG combined with HCG / LH-R upregulated the expression of VEGF gene and protein in endometrial cells.
【作者单位】：中山大学附属第一医院生殖中心;湖北医药学院附属人民医院生殖中心;
【基金】：国家自然科学基金资助项目(No:2008-30,872,762)
【分类号】：R714

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