VEGF和ADAMTS1在不育男性精浆中的表达及其与精液参数相关性分析

发布时间：2018-03-21 12:08

本文选题：不育　切入点：特发性弱精子症　出处：《郑州大学》2016年硕士论文　论文类型：学位论文

【摘要】：背景和目的不育症目前已发展成为了全球性问题,世界卫生组织(World Health Organization,WHO)的调查数据显示,在不孕不育夫妇中,50%以上存在男方生殖功能缺陷。导致男性不育的因素众多,其中若仅表现为精液参数中前向运动(Progressive motility,PR)精子的百分率32%且病因不明者,称为特发性弱精子症(idiopathic asthenospermia)。目前,特发性弱精子症的病因研究仍是生殖医学中的热点和难点。人类精浆中存在多种蛋白酶和细胞因子,它们对精子的运动能力有着一定的调节作用。血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)是一种分泌型的糖基化蛋白,由两个分子量为17-22KD大小的多肽链以二硫键的方式连接形成,其分子本身具有高度保守性,对热和酸稳定。VEGF是人体内的一种多功能细胞因子,能够强有力地刺激血管内皮细胞增生及血管生成。VEGF在人类的生殖系统中,通过受体介导发挥其生物学效应,促进生殖系统中各器官血管的生成,并参与调节血管通透性。在大鼠精子的顶体、尾部颈段、中段及主段均发现了VEGF受体Flt-1和KDR蛋白的表达,说明精子本身即为VEGF的靶细胞。VEGF在男性精浆中也有一定程度的表达,且当精液中的VEGF水平较高时,其女方的妊娠率也会相应增加。ADAMTS1(a disintegrin-like and metalloproteinase with thrombospond in type 1motifs)即含Ι型血小板结合蛋白基序的解聚蛋白样金属蛋白酶,它是一类广泛存在于细胞外基质(extracellularmatrix,ecm)中的zn2+依赖的分泌型金属蛋白酶,它可以通过降解brevican、versican和aggrecan等蛋白聚糖分子来参与ecm的重构,在人体的卵巢、子宫及泌尿生殖系统中均有不同程度的表达。adamts1可通过阻断vegf受体磷酸化来抑制内皮增殖和血管新生。同时,adamts1还可活化基质糖蛋白thrombospondin(tsp),而后者有抗血管再生的作用。agarwal等人发现,弱精子症患者精浆中的adamts1的表达水平较高于正常男性。s.e.aydos等人的研究也指出,精子的数量与活力均与精浆中adamts1的表达水平呈负相关。adamts1的重要功能之一是抑制血管新生,而vegf是目前体内所知的刺激血管新生的最强因子,但两者的表达是否相关,迄今尚不完全明确,基于它们对男性生殖系统的调节都有着重要意义,推测精浆中vegf、adamts1表达水平的改变可能影响了精子活力。本研究采用酶联免疫吸附法检测精浆中vegf、adamts1的表达水平,并分别将其与精液参数和助孕治疗结局进行相关性分析,探讨人类精浆中vegf、adamts1的表达水平对精子活力的影响。材料与方法所选取的研究对象为2015年11月-2016年1月期间在郑州大学第一附属医院生殖医学中心就诊行首次夫精人工授精(artificalinseminationbyhusband’ssperm,aih)助孕治疗的不育患者。其中,实验组为36名特发性弱精子症患者,正常对照组为31名精液分析各项参数正常且既往有生育史的男性,收集新鲜精液,行常规精液分析后离心沉淀,吸取上层精浆,采用酶联免疫吸附法检测两组患者精浆中vegf、adamts1的表达水平。结果1.特发性弱精子症组与正常对照组精浆中vegf的表达水平分别为:77.51±12.47pg/ml、147.05±15.26pg/ml,特发性弱精子症组与正常对照组相比较,精浆中vegf的表达水平下降,差异有统计学意义(p0.01)。2.特发性弱精子症组与正常对照组精浆中adamts1的表达水平分别为:92.38±16.77ng/ml、46.87±13.12ng/ml,特发性弱精子症组与正常对照组相比较,精浆中adamts1的表达水平升高,差异有统计学意义(p0.01)。3.特发性弱精子症组与正常对照组精浆中VEGF的表达水平与ADAMTS1的表达水平呈负相关(r=-0.433,P0.01)。4.特发性弱精子症组与正常对照组精浆中VEGF的表达水平与精液优化前的前向运动精子百分率呈正相关(r=0.741,P0.01)。5.特发性弱精子症组与正常对照组精浆中ADAMTS1的表达水平与精液优化前的前向运动精子百分率呈负相关(r=-0.347,P0.01)。6.特发性弱精子症组和正常对照组的临床妊娠率分别为11.11%、12.90%,差异无统计学意义(P0.05)。7.特发性弱精子症组和正常对照组精浆中的VEGF和ADAMTS1的表达水平与精液优化后的前向运动精子百分率无明显相关性(P0.05),二者与临床妊娠率均无明显相关性(P0.05)。结论1.精子活力与精浆中VEGF、ADAMTS1的表达水平有关,特发性弱精子症患者精子活力下降与VEGF降低、ADAMTS1升高密切相关。2.精液优化后精子活力、临床妊娠率与精浆中VEGF、ADAMTS1的表达水平无关。
[Abstract]:Background and objective: infertility has become a global problem, WHO (World Health Organization, WHO) survey data show that in infertile couples, more than 50% of the existence of defects. The reproductive function of the man many factors leading to male infertility, which showed only if the semen parameters of the forward motion (Progressive motility, PR) sperm the percentage of 32% and of unknown etiology, called idiopathic asthenospermia (idiopathic asthenospermia). At present, idiopathic asthenospermia in reproductive medicine etiology research is still hot and difficult. There are many proteases and cytokines in human seminal plasma, has a moderating effect on sperm motility the ability of vascular endothelial growth factor (vascular endothelial, growth factor, VEGF) is a glycosylated protein secreted by two, the molecular weight of the polypeptide chain of 17-22KD size The two way to connect the formation of disulfide bond, the molecule itself is highly conservative, stable to heat and acid.VEGF is a multifunctional cytokine in the human body, can strongly stimulate the proliferation of vascular endothelial cells and formation of.VEGF in the human reproductive system, mediated by the receptor exerts its biological effect, promote the formation of the vessels the reproductive organs of the system, and is involved in the regulation of vascular permeability in rats. The acrosome of sperm tail, cervical, middle and primary segments were found in the expression of VEGF receptor Flt-1 and KDR protein, that sperm itself is the target cell.VEGF VEGF also expresses in the seminal plasma, and when semen in the higher VEGF level, the woman's pregnancy rate will be a corresponding increase in.ADAMTS1 (a disintegrin-like and metalloproteinase with thrombospond in type 1motifs) including type 1 platelet binding protein motif A disintegrin and metalloprotease, which are widely distributed in the extracellular matrix (extracellularmatrix, ECM) in the zn2+ dependent secretory metalloprotease, it can be degraded by brevican, versican and aggrecan proteoglycan molecules to participate in the reconstruction of ECM in the human body, the ovary,.Adamts1 was expressed in uterus and genitourinary system in different degrees can inhibit the endothelial cell proliferation and angiogenesis by blocking VEGF receptor phosphorylation. At the same time, ADAMTS1 can also be activated matrix glycoprotein thrombospondin (TSP), while the latter has anti angiogenesis for.Agarwal et al found that the expression level of asthenospermia seminal plasma in ADAMTS1 was higher than that of normal male.S.e.aydos et al. Also pointed out that the quantity and vitality of sperm and the expression level of ADAMTS1 in seminal plasma is one of the important functions of.Adamts1 is negatively related to inhibition of angiogenesis, which is VEGF The strongest factor to stimulate angiogenesis in vivo is known, but the expression is related, so far is still not completely clear, based on their regulation on male reproductive system are of great significance, suggesting that VEGF in seminal plasma, and the change of ADAMTS1 expression may affect sperm motility. In this study, using enzyme-linked immunosorbent adsorption VEGF detection of seminal plasma, the expression level of ADAMTS1, and the semen parameters and pregnancy outcomes were analyzed respectively, to investigate the effect of VEGF in human seminal plasma, the expression level of ADAMTS1 on sperm motility. Materials and methods: the research object selected for the November 2015 -2016 year in January in the hospital for reproductive medicine center the First Affiliated Hospital of Zhengzhou University for the first time, artificial insemination (artificalinseminationbyhusband ssperm AIH) assisted reproductive treatment of infertile patients. Among them, the experimental group of 36 idiopathic asthenospermia In patients with normal control group, the parameters for normal and previous reproductive history of 31 male semen analysis, fresh semen was collected for routine semen analysis, after centrifugation, learn from the upper seminal plasma were detected by enzyme-linked immunosorbent assay in two patients in the seminal plasma of VEGF, ADAMTS1 expression. Results 1. idiopathic of asthenospermia group and normal control group, the expression level of VEGF in seminal plasma were 77.51 + 12.47pg/ml, 147.05 + 15.26pg/ml, idiopathic asthenospermia group compared with the normal control group, the expression level of VEGF in seminal plasma decreased, the difference was statistically significant (P0.01.2.) of idiopathic asthenospermia disease group and normal control group, the expression level of ADAMTS1 in seminal plasma were 92.38 + 16.77ng/ml, 46.87 + 13.12ng/ml, idiopathic asthenospermia group compared with the normal control group, the high expression level of ADAMTS1 in seminal plasma, there was significant difference (P0.01) of idiopathic.3. Asthenospermia group was negatively correlated with the expression level of the normal control group VEGF and the expression level of ADAMTS1 in seminal plasma of (r=-0.433, P0.01).4. idiopathic asthenospermia group and normal control expression level and semen optimization of VEGF group in the seminal plasma before the percentage of motile sperm was positive (r=0.741, P0.01).5. idiopathic asthenospermia group and normal control group ADAMTS1 expression level and semen optimization in seminal plasma before the sperm forward movement was negatively correlated (r=-0.347, P0.01).6. idiopathic weak sperm disease clinical group and normal control group, the pregnancy rate was 11.11%, 12.90%, the difference was not statistically significant (P0.05).7. idiopathic asthenospermia group and normal control group in seminal plasma VEGF and the expression of ADAMTS1 and semen after optimization of sperm forward movement had no significant correlation (P0.05), and the clinical pregnancy rate of two were not significantly correlated (P0.05). 1., sperm motility is related to the expression level of VEGF and ADAMTS1 in seminal plasma. The decrease of sperm motility and VEGF decrease and ADAMTS1 increase are closely related to sperm motility in patients with idiopathic asthenospermia. Sperm motility and clinical pregnancy rate after.2. semen optimization are not related to the expression level of VEGF and ADAMTS1 in seminal plasma.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R698.2

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