IL-1β和leptin对子宫内膜细胞分泌ICAM-1和MMP9的影响

发布时间：2018-07-10 09:00

本文选题：白细胞介素-1β + 瘦素　；参考：《青岛大学》2011年硕士论文

【摘要】：目的通过研究不同浓度的IL-1β(Interleukin-1β, IL-1β)和瘦素(leptin)对子宫内膜细胞所分泌的与胚胎着床密切相关的细胞间粘附分子-1 (Intercellular Adhesion Mo-lecule-1, ICAM-1)和基质金属蛋白酶9(Matrix Metalloproteinase 9, MMP9)表达的影响,来探讨IL-1β和leptin对胚胎着床的影响,为提高胚胎种植率提供理论依据。方法取育龄妇女分泌中期的子宫内膜进行体外培养,分别用不同浓度的IL-1β、leptin干预,采用免疫细胞化学方法明确ICAM-1和MMP9在子宫内膜腺体细胞或基质细胞的定位表达；采用ELISA方法测定干预后培养液中ICAM-1和MMP9的表达,并用半定量RT-PCR方法检测子宫内膜细胞中IC AM-1 mRNA和MMP9mRNA的表达。结果(1)免疫细胞化学：所培养的细胞为子宫内膜来源的细胞(包括腺体细胞和基质细胞),并且发现不论干预前后,ICAM-1主要在腺体细胞中表达,而基质细胞只有很少的表达,而MMP9在基质和腺体细胞中均有表达；(2) ELISA:与对照组相比,ICAM-1经lOng/ml的IL-1β干预后表达上调(P0.05),而分别经0.1ng/ml、1ng/ml的IL-1β干预后表达无显著性差异(P0.05)；MMP9分别经1ng/ml、lOng/ml的IL-1β干预后表达上调(P0.05),而经0.1ng/ml的IL-1β干预后表达无显著性差异(P0.05)。ICAM-1经100ng/ml的leptin干预后表达上调(P0.05),而经lng/ml、lOng/ml的leptin干预后表达无显著性差异(P0.05);MMP9分别经1ng/ml、10ng/ml、100ng/ml的leptin干预后表达均上调(P0.05)。(3) RT-PCR:0.1ng/ml IL-1β干预后ICAM-1 mRNA的表达量与对照组无差异(P0.05), 1ng/ml IL-1β、10ng/ml IL-1β干预后,与对照组相比,ICAM-1 mRNA的表达增加,且随浓度的升高,表达量呈上升趋势(P0.05),分别经不同浓度的IL-1β(0.1ng/ml、1ng/ml、10ng/ml), MMP9 mRNA较对照组相比表达均增加,且随浓度的升高,表达量呈上升趋势(P0.05)。而经不同浓度的leptin(1ng/ml、10ng/ml、100ng/ml)干预后,ICAM-1、MMP9 mRNA较对照组相比表达均增加,且随浓度的升高,表达量呈上升趋势(P0.05)。结论一定量的IL-1β和leptin可以增加体外培养分泌中期子宫内膜细胞ICAM-1和MMP9表达,且有剂量依赖性。
[Abstract]:Objective to investigate the effects of different concentrations of IL-1 尾 and leptin (leptin) on the expression of intercellular Adhesion Molecule-1 (ICAM-1) and matrix metalloproteinase 9 (MMP9) secreted by endometrial cells. To explore the effect of IL-1 尾 and leptin on embryo implantation and provide theoretical basis for increasing embryo implantation rate. Methods the endometrium of women of childbearing age was cultured in vitro, and the expression of ICAM-1 and MMP9 in endometrial glandular cells or stromal cells was determined by immunocytochemistry with different concentrations of IL-1 尾 -leptin. The expression of ICAM-1 and MMP9 was detected by Elisa and the expression of ICAM-1 mRNA and MMP9 mRNA in endometrial cells was detected by semi-quantitative RT-PCR. Results (1) Immunocytochemistry: the cultured cells were endometrial cells (including glandular cells and stromal cells), and it was found that ICAM-1 was mainly expressed in glandular cells before and after intervention, but only a little in stromal cells. The expression of MMP9 was up-regulated in stromal and glandular cells (P0.05) compared with control group (P0.05), but there was no significant difference after 0.1 ng / ml IL-1 尾 (P0.05). The expression of MMP9 was upregulated after 1 ng / ml of IL-1 尾 (P0.05), but there was no significant difference after the intervention of IL-1 尾 of 0.1ng/ml (P0.05). ICAM-1 was up-regulated by leptin of 100ng/ml (P0.05), but there was no significant difference in the expression of MMP9 after the intervention of leptin (P0.05). The expression of ICAM-1 mRNA was up-regulated (P0.05). (3). The expression of ICAM-1 mRNA was not different between the control group and the control group (P0.05) after the intervention of RT-PCR: 0.1ng / ml IL-1 尾 (P0.05). The expression of ICAM-1 mRNA was not different after the intervention of 1ng/ml IL-1 尾 10 ng / ml IL-1 尾. Compared with the control group, the expression of ICAM-1 mRNA increased, and the expression of ICAM-1 mRNA increased with the increase of the concentration (P0.05). After different concentrations of IL-1 尾 (0.1 ng / ml 1 ng / ml), the expression of MMP9 mRNA increased compared with the control group, and the expression of MMP9 mRNA increased with the increase of the concentration (P0.05). The expression of ICAM-1mMP9 was increased after different concentrations of leptin (1ng / ml 10ng / ml) compared with the control group, and the expression increased with the increase of the concentration (P0.05). Conclusion IL-1 尾 and leptin can increase the expression of ICAM-1 and MMP9 in endometrial cells in vitro in a dose-dependent manner.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R321

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