生长激素对薄型内膜患者临床结局的影响及机制探究

发布时间：2018-03-23 02:17

本文选题：生长激素　切入点：薄型内膜　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:在辅助生殖技术(Assisted Reproductive Technology,ART)中,子宫内膜厚度是影响妊娠结局的重要独立因素,多项研究显示HCG注射日内膜厚度与妊娠率正相关。对于薄型子宫内膜目前尚无公认确切有效的治疗方法。在我们临床工作中发现对薄型内膜患者应用生长激素(growth hormone,GH)可增厚内膜厚度并改善患者妊娠结局。有研究表明,内膜腺上皮中GH受体在种植窗期表达上调,这说明GH不仅通过增厚内膜的方式,还可能在胚胎种植过程发挥作用来改善妊娠结局。本研究回顾性分析临床数据,并通过体外培养的方法探讨GH对RL95-2细胞增殖、血管化及容受性指标的影响,分析可能机制。方法:回顾性分析93例薄型内膜患者数据,其中40例应用GH,53例未添加GH,GH添加方法为内膜增殖期加用GH 5IU/日,比较两组移植日内膜厚度、胚胎种植率和临床妊娠率等指标,并将GH组既往周期与添加GH周期最大内膜厚度进行比较;体外培养RL95-2细胞,检测GH对细胞增殖的影响(包括行CCK-8检测细胞增殖、流式细胞术检测细胞周期),并检测GH对细胞血管化及容受性指标的影响(行实时荧光定量PCR、Western blot检测所选取得8个与种植相关的容受性分子基因及蛋白表达)。结果:1.GH组较对照组移植日内膜厚度、胚胎种植率和临床妊娠率显著增加;2.GH组在当周期添加GH后最大内膜厚度显著高于既往未添加GH周期的最大内膜厚度;3.GH可使G1期减少、S期增加,促进细胞增殖;4.实时荧光定量PCR中GH作用后VEGF、Itg B3、IGF-I三者m RNA显著上调;5.Western blot中GH作用后VEGF、Itg B3、IGF-I三者蛋白表达增加;6.GH促进细胞增殖、促进VEGF、Itg B3、IGF-I三者基因及蛋白表达的作用均可被JAK/STAT通路抑制剂AG490抑制。7.GH对Itg AV、LIF、EGF、HOXA10、SPP1基因及蛋白表达无明显影响。结论:FET周期中添加GH可能可以增加薄型内膜者内膜厚度并改善其妊娠结局;GH可促进RL95-2细胞增殖、血管化指标VEGF及容受性分子Itg B3及IGF-I的表达,且GH改善薄型内膜者内膜厚度机制很可能与JAK/STAT通路有关。
[Abstract]:Objective: in assisted Reproductive assisted technique (art), endometrial thickness is an important independent factor affecting pregnancy outcome. A number of studies have shown that the thickness of endometrium on the day of HCG injection is positively correlated with pregnancy rate. There is no known effective treatment for thin endometrium. In our clinical work, growth hormone growth was found in patients with thin endometrium. Hormoney GHB can increase intimal thickness and improve pregnancy outcomes in patients. The expression of GH receptor in endometrial glandular epithelium is up-regulated during implantation window, which indicates that GH may play a role in improving pregnancy outcome not only by thickening the intima, but also during embryo implantation. The effects of GH on proliferation, vascularization and receptivity of RL95-2 cells were studied in vitro. Methods: the data of 93 patients with thin intima were analyzed retrospectively. Among them, 40 cases were treated with GH 5IU/ during endometrial proliferation. The thickness of endometrium, embryo implantation rate and clinical pregnancy rate were compared between the two groups. The effects of GH on the proliferation of RL95-2 cells were measured by CCK-8. Flow cytometry was used to detect cell cycle, and the effects of GH on cell vascularization and receptivity were detected. The expression of eight receptive genes and proteins related to implantation was obtained by real-time fluorescence quantitative PCR Western blot. The thickness of endometrium in GH group was higher than that in control group. The embryo implantation rate and clinical pregnancy rate were significantly increased. 2. The maximum intimal thickness of GH group was significantly higher than that of no GH cycle. 3. GH decreased the increase of S phase in G1 phase. The expression of VEGFG Itg B3IGF-I in real-time fluorescent quantitative PCR was significantly up-regulated, and the expression of VEGF Itg B3 IGF-I protein increased significantly after GH treatment in Western blot. 6. GH promoted cell proliferation, and increased the expression of VEGFG B3IGF-I protein in the cell proliferation of VEGFG B3IGF-I, and increased the expression of IGF-I protein in VEGFG B3IGF-I. The effects of VEGFG B3IGF-I gene and protein expression can be inhibited by JAK/STAT pathway inhibitor AG490. 7. GH has no significant effect on the expression of SPP1 gene and protein in Itg AVLIFEGFIFFHXA10. Conclusion the addition of GH in the JAK/STAT cycle may increase the intimal thickness and increase the intimal thickness of the thin endometrium. Improving the outcome of pregnancy can promote the proliferation of RL95-2 cells. The expression of vascularization index VEGF, receptive molecule Itg B3 and IGF-I, and the mechanism of GH improving intimal thickness of thin endometrium may be related to JAK/STAT pathway.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.8

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