本文選題：kallistatin + 男性不育癥�。� 參考：《昆明理工大學》2017年碩士論文

【摘要】：目的:不孕不育癥已成為影響育齡期夫妻健康與生活的重要疾病。隨著生殖醫(yī)學的發(fā)展與人們觀念的改變,男性不育癥的問題越來越多的關注。但男性不育癥的病因復雜,病理機制尚未完全明確,并且目前無特效治療藥物,探索新的治療靶點就成為當下的研究熱點之一。男性生精功能是生殖功能中極為重要的一部分,精子的密度、活動力、形態(tài)等決定著精子能否正常存活與能否完成受精,而精子的生成是一系列復雜而有序的變化過程。激肽釋放酶一激肽系統(tǒng)(kallikrein-kinin system,KKS)是人體內重要的炎性調節(jié)系統(tǒng),研究發(fā)現(xiàn)其可能在精子發(fā)生過程中具有調控作用。該系統(tǒng)中組織激肽釋放酶結合蛋白(kallistatin,KS)在體內分布廣泛,在多個系統(tǒng)和多種疾病中發(fā)揮著抗氧化應激、抗纖維化、舒張血管、抗血管生成、抗凋亡、抗腫瘤、脂代謝調節(jié)等多重生物學效應,在前期研究中我們發(fā)現(xiàn),kallistatin基因與大鼠生精功能具有相關性,kallistatin轉基因大鼠睪丸組織生精功能顯著增強。因此我們推測kallistatin亦能通過睪丸局部血管舒張效應、抗氧化、抗凋亡、抗纖維化以及組織損傷修復等發(fā)揮男性生殖功能保護作用,因此本研究將通過探討kallistatin在男性不育患者精漿和睪丸組織以及不同生理年齡SD大鼠睪丸組織中的表達水平,以明確其與男性不育癥的關系,為探討kallistatin能否成為男性不育癥新的治療靶點奠定基礎。方法:收集2015年10月至2017年2月在云南省第一人民醫(yī)院生殖醫(yī)學科就診的40例男性不育患者精液標本,和10例正常精液標本,通過ELISA技術檢測精漿中kallistatin蛋白的表達水平,初步探討kallistatin與男性不育癥的相關性;同時收集不同類型男性不育癥患者(正常精子、梗阻性無精子癥、非梗阻性無精子癥和嚴重少弱畸精子癥)睪丸組織40例以及8個不同生理年齡組SD大鼠(4周、8周、12周、20周、30周、40周、60周、80周)睪丸組織共80例,通過RT-PCR、Western blot和免疫組織化學技術檢測kallistatin表達情況,以進一步明確kallistatin在生精功能中的作用。結果:kallistatin蛋白在男性不育癥患者精漿中的表達水平低于正常對照組,少精子癥患者精漿中的表達水平顯著低于正常對照組(42.5%)。kallistatin在不同類型男性不育癥患者睪丸組織中表達水平依次為:正常對照組多梗阻性無精子癥嚴重少弱畸精子癥非梗阻性無精子癥(P0.05)。kallistatin在不同年齡段的大鼠睪丸中均有表達,表達量與大鼠年齡增長相關,幼年期表達量較低,后隨年齡增長逐漸增高,在20周左右達到峰值并維持較高水平至40周,后隨年齡增長而降低。結論:通過本課題研究發(fā)現(xiàn),kallistatin與男性不育癥有相關性,在少精癥患者精漿中以及非梗阻性無精癥患者睪丸中kallistatin的表達水平均顯著降低,提示kallistatin可能與生精功能相關。kallistatin在不同年齡大鼠睪丸組織中的表達量與大鼠年齡改變具有相關性,在幼齡期較低,隨后逐漸增高,從進入繁殖期開始表達量達到高峰,并維持到最大生育年齡中期,后隨年齡增長下降至較低水平,提示kallistatin可能在男性生殖功能發(fā)展與衰退中起到調控作用。
[Abstract]:Objective: infertility has become important diseases affecting the health and life of childbearing age couples. With the development of reproductive medicine and changing people's idea of male infertility problem more and more attention. But the cause of male infertility complicated pathological mechanism is not completely clear, and there is no specific treatment, to explore a new therapeutic target for it has become a hot research topic at the moment. Male spermatogenesis is one of the most important part of reproductive function, sperm density, motility, morphology determines whether the normal sperm survival and fertilization can be completed, and the generation of sperm is a series of complicated and orderly process. The kallikrein kinin system (kallikrein-kinin system KKS) is an important inflammatory regulation system in the human body, the study found that may have a regulatory role in spermatogenesis. The tissue kallikrein system Binding proteins (kallistatin, KS) are widely distributed in the body in multiple systems and a variety of diseases play anti oxidative stress, anti fibrosis, vasorelaxation, anti angiogenesis, anti apoptosis, anti tumor, lipid metabolism and other multiple biological effects, we found in the previous study, the kallistatin gene is associated with spermatogenesis the function of rat testis kallistatin transgenic rat spermatogenic function significantly enhanced. So we speculate that kallistatin can also by testicular local vasodilating effects, antioxidation, anti apoptosis, anti fibrosis and tissue repair may play a protective role in male reproductive function, this study will explore through the expression level of kallistatin in seminal plasma of infertile patients and testis tissue and different physiological age of SD rat testis tissue and to clarify its relationship with male infertility, to investigate whether kallistatin will become a man Lay the foundation for a new target in the treatment of infertility. Methods: from October 2015 to February 2017 in 40 cases of male infertility patients with semen samples of first people's Hospital of Yunnan Province Department of reproductive medicine, and 10 normal semen samples, the expression level of kallistatin protein detection precision ELISA technology in pulp, preliminary Study on correlation between kallistatin and male infertility; at the same time to collect different types of patients with male infertility (normal sperm, obstructive azoospermia, non obstructive azoospermia and severe oligoasthenoteratozoospermia) of testicular tissue in 40 cases and 8 different physiological age groups of SD rats (4 weeks, 8 weeks, 12 weeks, 20 weeks, 30 weeks, 40 weeks, 60 weeks, 80 week) a total of 80 cases of testicular tissue, by RT-PCR, Western blot and immunohistochemistry to detect the expression of kallistatin, in order to further clarify the kallistatin in spermatogenic function. Results: kallistatin protein in male The expression level of seminal in lower than normal control group, the expression level of semen of oligospermia patients was significantly lower than that in normal control group (42.5%).Kallistatin expression in testicular tissue of patients with different types of male infertility are: normal control group of obstructive azoospermia severe oligoasthenoteratozoospermia (non obstructive azoospermia P0.05.Kallistatin) in different age rat testis tissues. The expression of rat age related to childhood low expression level, after gradually increased with age, reached a peak in 20 weeks and 40 weeks to maintain a high level, then decreased with the increasing of age. Conclusion: through this the research found that the relationship between kallistatin and male infertility, sperm in oligospermic candidates and non obstructive azoospermia patients with testicular kallistatin expression level was significantly decreased, suggesting that K Allistatin may be related to spermatogenesis related.Kallistatin expression in different age rat testis in rats and correlation with age changes in the young, low, then increased gradually, from entering the breeding period began the expression reached the peak, and to maintain the maximum fertility age after the mid low with the increase of age the level, suggesting that kallistatin may play a regulatory role in male reproductive function development and decline.

【學位授予單位】：昆明理工大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R698.2

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