本文選題：維生素D　切入點(diǎn)：精子活力　出處：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的在世界范圍內(nèi),男性不育占不孕不育總?cè)藬?shù)的40-50%,它影響大約7%的所有男性。在男性不育的眾多原因中,維生素D被認(rèn)為與男性的生育能力有直接關(guān)系。本研究旨在探索維生素D與男性精液質(zhì)量的關(guān)系,為研究維生素D對(duì)男性生殖的具體作用機(jī)制提供更全面的研究方向。并在體外以維生素D的活性物質(zhì)--1,25(OH)2D3處理精子,研究其對(duì)精子的作用機(jī)制。方法第一部分血清和精漿維生素D與精子參數(shù)的臨床相關(guān)性分析根據(jù)納入標(biāo)準(zhǔn)和排除標(biāo)準(zhǔn)收集我院生殖醫(yī)學(xué)中心男科門(mén)診就診的男性患者以及正常體檢者精液標(biāo)本,按WHO《人類(lèi)精液檢查與處理實(shí)驗(yàn)室手冊(cè)》(第五版)標(biāo)準(zhǔn),應(yīng)用WLJY-9000型偉力彩色精子質(zhì)量檢測(cè)系統(tǒng)檢測(cè)精子濃度、精子總數(shù)、精子總活率、前向運(yùn)動(dòng)精子百分率以及相應(yīng)運(yùn)動(dòng)學(xué)參數(shù)[曲線速度(VCL)、直線速度(VSL)、平均路徑速度(VAP)];吖啶橙染色法檢測(cè)精子DNA碎片指數(shù)(DFI)和高DNA可染性(HDS);應(yīng)用改良巴氏染色法以WHO標(biāo)準(zhǔn)嚴(yán)格判讀精子形態(tài),評(píng)估正常形態(tài)精子百分率;應(yīng)用電化學(xué)發(fā)光法檢測(cè)血清和精漿維生素D水平。分析血清和精漿維生素D與精液各參數(shù)之間的相關(guān)性。第二部分1,25(OH)2D3對(duì)精子運(yùn)動(dòng)能力的影響及其作用機(jī)制根據(jù)納入標(biāo)準(zhǔn)和排除標(biāo)準(zhǔn)收集我院生殖醫(yī)學(xué)中心男科門(mén)診就診的男性患者以及正常體檢者的精液標(biāo)本,用percoll液密度梯度離心法提取納入研究人群的精子,以不同濃度(0、0.01nM、0.1nM、1nM、10nM)的 1,25(OH)2D3處理精子,在不同時(shí)間點(diǎn)(0、15min、30min、60min、90min)用WLJY-9000型偉力彩色精子質(zhì)量檢測(cè)系統(tǒng)檢測(cè)精子各運(yùn)動(dòng)參數(shù);根據(jù)前期結(jié)果,選擇最適濃度的1,25(OH)2D3處理精子30min后,檢測(cè)精子的爬高能力、cAMP的含量以及精子PKA的活性、NADH和ATP的含量,分析各指標(biāo)的變化。結(jié)果1.臨床試驗(yàn)中,血清維生素D的水平與精子各參數(shù)之間無(wú)相關(guān)性(p0.05);精漿維生素D與精子總活率(r = 0.308,p = 0.026)、前向運(yùn)動(dòng)精子百分率(r= 0.299,p = 0.031)、曲線速度(r = 0.386,p = 0.005)、直線速度(r = 0.348,p = 0.011)、路徑速度(r = 0.383,p = 0.005)之間呈顯著正相關(guān)。2.1,25(OH)2D3處理精子后,在作用30分鐘且濃度為0.1nM時(shí),精子的各項(xiàng)運(yùn)動(dòng)參數(shù)增加顯著(p0.001),精子的爬高高度顯著增加(p0.01),精子NADH、ATP和cAMP含量顯著增加(P0.05),精子PKA活性顯著增強(qiáng)(p0.05)。結(jié)論1.精漿維生素D可能參與精子運(yùn)動(dòng)能力的調(diào)節(jié)。2.活性形式的維生素D可能通過(guò)cAMP/PKA和NADH途徑促進(jìn)精子線粒體電子呼吸鏈的作用,增加ATP的產(chǎn)量,從而增強(qiáng)精子的活力。
[Abstract]:Objective in the world, male infertility accounts for 40-50% of the total number of infertile women, which affects about 7% of all men.Among the many causes of male infertility, vitamin D is thought to be directly related to men's fertility.The purpose of this study was to explore the relationship between vitamin D and male semen quality, and to provide a more comprehensive research direction for the study of the specific mechanism of vitamin D on male reproduction.The spermatozoa were treated with the active substance of vitamin D in vitro, and the mechanism of its action on spermatozoa was studied.Methods the clinical correlation between serum and seminal plasma vitamin D and sperm parameters was analyzed. According to the inclusion and exclusion criteria, semen samples were collected from male patients in andrology clinic and normal persons in our center of reproductive medicine.According to the WHO Manual for the Laboratory of Human semen examination and processing (fifth edition), the sperm concentration, total sperm count and total sperm motility were detected by using the WLJY-9000 type Weili color sperm quality detection system.Forward motility sperm percentage and its corresponding kinematic parameters [curve velocities, linear velocities, VSLs, average path velocities]; acridine orange staining for the detection of sperm DNA fragment index (DNA) and high DNA dyeability (WHO); using modified pasteurian staining method to detect WHOThe standard strictly interprets sperm morphology,The percentage of normal spermatozoa was evaluated and the levels of vitamin D in serum and seminal plasma were measured by electrochemiluminescence (ECL).The correlation between serum and seminal plasma vitamin D and semen parameters was analyzed.The second part: the effect of OHH 2D3 on sperm motility and its mechanism of action. According to the inclusion and exclusion criteria, semen samples were collected from male patients from andrology outpatient clinics of our Reproductive Medicine Center, as well as normal physical examination subjects.The content of camp in sperm and the content of nadh and ATP in sperm PKA were determined after the treatment of 30min with the optimum concentration of 1o 25 OHH 2D3, and the changes of each index were analyzed.Result 1.In clinical trials,There was a significant positive correlation between path velocities (r = 0.383, p = 0.005).At the concentration of 0.1nM for 30 minutes, the motility parameters of spermatozoa increased significantly (p0.001), the climbing height of spermatozoa increased significantly (p0.01), the contents of NADH-ATP and cAMP increased significantly, and the activity of PKA increased significantly (p0.05).Conclusion 1.Seminal plasma vitamin D may be involved in the regulation of sperm motility.The active form of vitamin D may promote the action of mitochondrial electron respiratory chain through cAMP/PKA and NADH pathway, and increase the production of ATP, thus enhancing sperm motility.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R698.2

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本文編號(hào)：1723272