發(fā)布時(shí)間：2018-03-23 02:17

  本文選題：生長激素　切入點(diǎn)：薄型內(nèi)膜　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:在輔助生殖技術(shù)(Assisted Reproductive Technology,ART)中,子宮內(nèi)膜厚度是影響妊娠結(jié)局的重要獨(dú)立因素,多項(xiàng)研究顯示HCG注射日內(nèi)膜厚度與妊娠率正相關(guān)。對(duì)于薄型子宮內(nèi)膜目前尚無公認(rèn)確切有效的治療方法。在我們臨床工作中發(fā)現(xiàn)對(duì)薄型內(nèi)膜患者應(yīng)用生長激素(growth hormone,GH)可增厚內(nèi)膜厚度并改善患者妊娠結(jié)局。有研究表明,內(nèi)膜腺上皮中GH受體在種植窗期表達(dá)上調(diào),這說明GH不僅通過增厚內(nèi)膜的方式,還可能在胚胎種植過程發(fā)揮作用來改善妊娠結(jié)局。本研究回顧性分析臨床數(shù)據(jù),并通過體外培養(yǎng)的方法探討GH對(duì)RL95-2細(xì)胞增殖、血管化及容受性指標(biāo)的影響,分析可能機(jī)制。方法:回顧性分析93例薄型內(nèi)膜患者數(shù)據(jù),其中40例應(yīng)用GH,53例未添加GH,GH添加方法為內(nèi)膜增殖期加用GH 5IU/日,比較兩組移植日內(nèi)膜厚度、胚胎種植率和臨床妊娠率等指標(biāo),并將GH組既往周期與添加GH周期最大內(nèi)膜厚度進(jìn)行比較;體外培養(yǎng)RL95-2細(xì)胞,檢測(cè)GH對(duì)細(xì)胞增殖的影響(包括行CCK-8檢測(cè)細(xì)胞增殖、流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期),并檢測(cè)GH對(duì)細(xì)胞血管化及容受性指標(biāo)的影響(行實(shí)時(shí)熒光定量PCR、Western blot檢測(cè)所選取得8個(gè)與種植相關(guān)的容受性分子基因及蛋白表達(dá))。結(jié)果:1.GH組較對(duì)照組移植日內(nèi)膜厚度、胚胎種植率和臨床妊娠率顯著增加;2.GH組在當(dāng)周期添加GH后最大內(nèi)膜厚度顯著高于既往未添加GH周期的最大內(nèi)膜厚度;3.GH可使G1期減少、S期增加,促進(jìn)細(xì)胞增殖;4.實(shí)時(shí)熒光定量PCR中GH作用后VEGF、Itg B3、IGF-I三者m RNA顯著上調(diào);5.Western blot中GH作用后VEGF、Itg B3、IGF-I三者蛋白表達(dá)增加;6.GH促進(jìn)細(xì)胞增殖、促進(jìn)VEGF、Itg B3、IGF-I三者基因及蛋白表達(dá)的作用均可被JAK/STAT通路抑制劑AG490抑制。7.GH對(duì)Itg AV、LIF、EGF、HOXA10、SPP1基因及蛋白表達(dá)無明顯影響。結(jié)論:FET周期中添加GH可能可以增加薄型內(nèi)膜者內(nèi)膜厚度并改善其妊娠結(jié)局;GH可促進(jìn)RL95-2細(xì)胞增殖、血管化指標(biāo)VEGF及容受性分子Itg B3及IGF-I的表達(dá),且GH改善薄型內(nèi)膜者內(nèi)膜厚度機(jī)制很可能與JAK/STAT通路有關(guān)。
[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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R714.8

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