本文選題：不育　切入點：特發(fā)性弱精子癥　出處：《鄭州大學》2016年碩士論文　論文類型：學位論文

【摘要】：背景和目的不育癥目前已發(fā)展成為了全球性問題,世界衛(wèi)生組織(World Health Organization,WHO)的調(diào)查數(shù)據(jù)顯示,在不孕不育夫婦中,50%以上存在男方生殖功能缺陷。導致男性不育的因素眾多,其中若僅表現(xiàn)為精液參數(shù)中前向運動(Progressive motility,PR)精子的百分率32%且病因不明者,稱為特發(fā)性弱精子癥(idiopathic asthenospermia)。目前,特發(fā)性弱精子癥的病因研究仍是生殖醫(yī)學中的熱點和難點。人類精漿中存在多種蛋白酶和細胞因子,它們對精子的運動能力有著一定的調(diào)節(jié)作用。血管內(nèi)皮細胞生長因子(vascular endothelial growth factor,VEGF)是一種分泌型的糖基化蛋白,由兩個分子量為17-22KD大小的多肽鏈以二硫鍵的方式連接形成,其分子本身具有高度保守性,對熱和酸穩(wěn)定。VEGF是人體內(nèi)的一種多功能細胞因子,能夠強有力地刺激血管內(nèi)皮細胞增生及血管生成。VEGF在人類的生殖系統(tǒng)中,通過受體介導發(fā)揮其生物學效應,促進生殖系統(tǒng)中各器官血管的生成,并參與調(diào)節(jié)血管通透性。在大鼠精子的頂體、尾部頸段、中段及主段均發(fā)現(xiàn)了VEGF受體Flt-1和KDR蛋白的表達,說明精子本身即為VEGF的靶細胞。VEGF在男性精漿中也有一定程度的表達,且當精液中的VEGF水平較高時,其女方的妊娠率也會相應增加。ADAMTS1(a disintegrin-like and metalloproteinase with thrombospond in type 1motifs)即含Ι型血小板結(jié)合蛋白基序的解聚蛋白樣金屬蛋白酶,它是一類廣泛存在于細胞外基質(zhì)(extracellularmatrix,ecm)中的zn2+依賴的分泌型金屬蛋白酶,它可以通過降解brevican、versican和aggrecan等蛋白聚糖分子來參與ecm的重構(gòu),在人體的卵巢、子宮及泌尿生殖系統(tǒng)中均有不同程度的表達。adamts1可通過阻斷vegf受體磷酸化來抑制內(nèi)皮增殖和血管新生。同時,adamts1還可活化基質(zhì)糖蛋白thrombospondin(tsp),而后者有抗血管再生的作用。agarwal等人發(fā)現(xiàn),弱精子癥患者精漿中的adamts1的表達水平較高于正常男性。s.e.aydos等人的研究也指出,精子的數(shù)量與活力均與精漿中adamts1的表達水平呈負相關(guān)。adamts1的重要功能之一是抑制血管新生,而vegf是目前體內(nèi)所知的刺激血管新生的最強因子,但兩者的表達是否相關(guān),迄今尚不完全明確,基于它們對男性生殖系統(tǒng)的調(diào)節(jié)都有著重要意義,推測精漿中vegf、adamts1表達水平的改變可能影響了精子活力。本研究采用酶聯(lián)免疫吸附法檢測精漿中vegf、adamts1的表達水平,并分別將其與精液參數(shù)和助孕治療結(jié)局進行相關(guān)性分析,探討人類精漿中vegf、adamts1的表達水平對精子活力的影響。材料與方法所選取的研究對象為2015年11月-2016年1月期間在鄭州大學第一附屬醫(yī)院生殖醫(yī)學中心就診行首次夫精人工授精(artificalinseminationbyhusband’ssperm,aih)助孕治療的不育患者。其中,實驗組為36名特發(fā)性弱精子癥患者,正常對照組為31名精液分析各項參數(shù)正常且既往有生育史的男性,收集新鮮精液,行常規(guī)精液分析后離心沉淀,吸取上層精漿,采用酶聯(lián)免疫吸附法檢測兩組患者精漿中vegf、adamts1的表達水平。結(jié)果1.特發(fā)性弱精子癥組與正常對照組精漿中vegf的表達水平分別為:77.51±12.47pg/ml、147.05±15.26pg/ml,特發(fā)性弱精子癥組與正常對照組相比較,精漿中vegf的表達水平下降,差異有統(tǒng)計學意義(p0.01)。2.特發(fā)性弱精子癥組與正常對照組精漿中adamts1的表達水平分別為:92.38±16.77ng/ml、46.87±13.12ng/ml,特發(fā)性弱精子癥組與正常對照組相比較,精漿中adamts1的表達水平升高,差異有統(tǒng)計學意義(p0.01)。3.特發(fā)性弱精子癥組與正常對照組精漿中VEGF的表達水平與ADAMTS1的表達水平呈負相關(guān)(r=-0.433,P0.01)。4.特發(fā)性弱精子癥組與正常對照組精漿中VEGF的表達水平與精液優(yōu)化前的前向運動精子百分率呈正相關(guān)(r=0.741,P0.01)。5.特發(fā)性弱精子癥組與正常對照組精漿中ADAMTS1的表達水平與精液優(yōu)化前的前向運動精子百分率呈負相關(guān)(r=-0.347,P0.01)。6.特發(fā)性弱精子癥組和正常對照組的臨床妊娠率分別為11.11%、12.90%,差異無統(tǒng)計學意義(P0.05)。7.特發(fā)性弱精子癥組和正常對照組精漿中的VEGF和ADAMTS1的表達水平與精液優(yōu)化后的前向運動精子百分率無明顯相關(guān)性(P0.05),二者與臨床妊娠率均無明顯相關(guān)性(P0.05)。結(jié)論1.精子活力與精漿中VEGF、ADAMTS1的表達水平有關(guān),特發(fā)性弱精子癥患者精子活力下降與VEGF降低、ADAMTS1升高密切相關(guān)。2.精液優(yōu)化后精子活力、臨床妊娠率與精漿中VEGF、ADAMTS1的表達水平無關(guān)。
[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.

【學位授予單位】：鄭州大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R698.2

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