kallistatin基因在男性不育症中的初步研究

发布时间：2018-04-13 08:55

本文选题：kallistatin + 男性不育症　；参考：《昆明理工大学》2017年硕士论文

【摘要】：目的:不孕不育症已成为影响育龄期夫妻健康与生活的重要疾病。随着生殖医学的发展与人们观念的改变,男性不育症的问题越来越多的关注。但男性不育症的病因复杂,病理机制尚未完全明确,并且目前无特效治疗药物,探索新的治疗靶点就成为当下的研究热点之一。男性生精功能是生殖功能中极为重要的一部分,精子的密度、活动力、形态等决定着精子能否正常存活与能否完成受精,而精子的生成是一系列复杂而有序的变化过程。激肽释放酶一激肽系统(kallikrein-kinin system,KKS)是人体内重要的炎性调节系统,研究发现其可能在精子发生过程中具有调控作用。该系统中组织激肽释放酶结合蛋白(kallistatin,KS)在体内分布广泛,在多个系统和多种疾病中发挥着抗氧化应激、抗纤维化、舒张血管、抗血管生成、抗凋亡、抗肿瘤、脂代谢调节等多重生物学效应,在前期研究中我们发现,kallistatin基因与大鼠生精功能具有相关性,kallistatin转基因大鼠睾丸组织生精功能显著增强。因此我们推测kallistatin亦能通过睾丸局部血管舒张效应、抗氧化、抗凋亡、抗纤维化以及组织损伤修复等发挥男性生殖功能保护作用,因此本研究将通过探讨kallistatin在男性不育患者精浆和睾丸组织以及不同生理年龄SD大鼠睾丸组织中的表达水平,以明确其与男性不育症的关系,为探讨kallistatin能否成为男性不育症新的治疗靶点奠定基础。方法:收集2015年10月至2017年2月在云南省第一人民医院生殖医学科就诊的40例男性不育患者精液标本,和10例正常精液标本,通过ELISA技术检测精浆中kallistatin蛋白的表达水平,初步探讨kallistatin与男性不育症的相关性;同时收集不同类型男性不育症患者(正常精子、梗阻性无精子症、非梗阻性无精子症和严重少弱畸精子症)睾丸组织40例以及8个不同生理年龄组SD大鼠(4周、8周、12周、20周、30周、40周、60周、80周)睾丸组织共80例,通过RT-PCR、Western blot和免疫组织化学技术检测kallistatin表达情况,以进一步明确kallistatin在生精功能中的作用。结果:kallistatin蛋白在男性不育症患者精浆中的表达水平低于正常对照组,少精子症患者精浆中的表达水平显著低于正常对照组(42.5%)。kallistatin在不同类型男性不育症患者睾丸组织中表达水平依次为:正常对照组多梗阻性无精子症严重少弱畸精子症非梗阻性无精子症(P0.05)。kallistatin在不同年龄段的大鼠睾丸中均有表达,表达量与大鼠年龄增长相关,幼年期表达量较低,后随年龄增长逐渐增高,在20周左右达到峰值并维持较高水平至40周,后随年龄增长而降低。结论:通过本课题研究发现,kallistatin与男性不育症有相关性,在少精症患者精浆中以及非梗阻性无精症患者睾丸中kallistatin的表达水平均显著降低,提示kallistatin可能与生精功能相关。kallistatin在不同年龄大鼠睾丸组织中的表达量与大鼠年龄改变具有相关性,在幼龄期较低,随后逐渐增高,从进入繁殖期开始表达量达到高峰,并维持到最大生育年龄中期,后随年龄增长下降至较低水平,提示kallistatin可能在男性生殖功能发展与衰退中起到调控作用。
[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.

【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R698.2

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