大豆異黃酮對(duì)男（雄）性精液質(zhì)量及性激素水平的影響
發(fā)布時(shí)間：2018-11-17 11:04
【摘要】：目的通過(guò)對(duì)重慶地區(qū)普通成年男性流行病學(xué)調(diào)查及實(shí)驗(yàn)室檢測(cè),了解該地區(qū)成年男性大豆異黃酮內(nèi)暴露水平,明確大豆異黃酮內(nèi)暴露水平與男性精液質(zhì)量和性激素水平之間的關(guān)聯(lián)。通過(guò)動(dòng)物實(shí)驗(yàn)研究探討不同劑量下的大豆異黃酮代謝物對(duì)血清性激素水平及精液質(zhì)量的影響。為探討大豆異黃酮對(duì)男性生殖健康的不良影響提供參考依據(jù)。方法1、人群研究:研究對(duì)象為2015年在重慶市豐都縣社區(qū)招募的普通成年男性,通過(guò)復(fù)合式問(wèn)卷收集研究對(duì)象的基本信息,現(xiàn)場(chǎng)采集志愿者的血液、尿液及精液樣本。采用高效液相色譜(High performance liquid chromatography,HPLC)分析人群尿液大豆異黃酮代謝物水平,計(jì)算機(jī)輔助精子分析系統(tǒng)(Computer-aided sperm analyzer,CASA)進(jìn)行精液參數(shù)分析,Diff-Quick試劑盒染色檢測(cè)精子形態(tài),染色質(zhì)結(jié)構(gòu)分析法(Sperm chromatin structure assay,SCSA)檢測(cè)精子核DNA碎片指數(shù),實(shí)時(shí)熒光定量PCR方法測(cè)定精子線粒體DNA(Mitochondrial DNA,mtDNA)拷貝數(shù),長(zhǎng)鏈PCR方法檢測(cè)mtDNA完整性,化學(xué)發(fā)光法檢測(cè)人群血清性激素水平(睪酮、卵泡刺激素、黃體生成素、雌二醇、孕酮、泌乳素)。采用SPSS16.0軟件分析了大豆異黃酮尿代謝物水平與精液質(zhì)量及性激素水平之間的關(guān)聯(lián)。研究對(duì)象的納入標(biāo)準(zhǔn)為:(1)禁欲天數(shù)為2~7天;(2)年齡18~55周歲男性;(3)無(wú)泌尿系統(tǒng)感染史、附睪炎病史、睪丸損傷疾病史、精索靜脈曲張治療史、睪丸下降不全史;(4)無(wú)喉結(jié)、陰毛、乳腺發(fā)育異常、陰莖異常、睪丸損傷、附睪炎癥、精索靜脈曲張、隱睪術(shù)后;(5)無(wú)五項(xiàng)精液參數(shù)均缺失者;(6)尿肌酐(Creatinine,CR)濃度在0.3~3g/L。最終納入161例作為研究對(duì)象。2、動(dòng)物實(shí)驗(yàn)研究:采用60只4周齡(青春期)SD雄性大鼠進(jìn)行亞慢性染毒,隨機(jī)分為6組,分別是正常對(duì)照組、染料木素5mg/kg、50mg/kg、500 mg/kg和2000mg/kg組、陽(yáng)性對(duì)照己烯雌酚0.1mg/kg組,連續(xù)灌胃29天。分別在染毒結(jié)束時(shí)(8周齡)以及動(dòng)物成年后(11周齡)進(jìn)行取材,觀察指標(biāo)包括臟器系數(shù)、睪丸組織結(jié)構(gòu)、性激素水平及精液質(zhì)量。采用蛋白質(zhì)印跡(westernblot,wb)檢測(cè)睪丸erβ的表達(dá)水平。結(jié)果1、人群研究:本次研究檢測(cè)了人群尿液染料木素、大豆苷元,黃豆黃素,雌馬酚,二氫大豆苷元代謝物水平。jonckheere-terpstra檢驗(yàn)結(jié)果顯示,隨著總的大豆異黃酮濃度等級(jí)的增加,精液體積和精子密度呈現(xiàn)遞減趨勢(shì),結(jié)果有統(tǒng)計(jì)學(xué)意義(p=0.044和0.018),隨著雌馬酚水平的增加,精液體積呈增加的趨勢(shì)(p=0.034)。在性激素水平上,染料木素與黃體生成素(p=0.019),黃豆黃素與睪酮(p㩳0.001),二氫大豆苷元與孕酮(p=0.021)均呈負(fù)相關(guān)。采用多元線性回歸方法校正潛在混雜因素(年齡、體質(zhì)指數(shù)、禁欲天數(shù)、吸煙狀態(tài)、飲酒狀態(tài))后,大豆異黃酮尿代謝物水平與上述精液參數(shù)關(guān)聯(lián)消失,與性激素水平關(guān)聯(lián)依舊存在。本研究未發(fā)現(xiàn)大豆異黃酮與dna損傷指標(biāo)(dna碎片指數(shù),mtdna拷貝數(shù),mtdna完整性)之間存在關(guān)聯(lián)。此外多元線性回歸結(jié)果顯示,精子dna損傷指標(biāo)與精液質(zhì)量之間存在關(guān)聯(lián)。2、動(dòng)物實(shí)驗(yàn)研究:大鼠青春期攝入染料木素后,染毒結(jié)束時(shí)及成年期染料木素各劑量組大鼠體重及臟器系數(shù)較正常對(duì)照組比,差異無(wú)統(tǒng)計(jì)學(xué)意義。he染色結(jié)果顯示染毒結(jié)束時(shí)染料木素2000mg/kg組睪丸組織生精小管細(xì)胞管腔中細(xì)胞稀疏,排列紊亂,成年期染料木素2000mg/kg組大鼠生精小管管腔內(nèi)精子數(shù)量較正常對(duì)照組比減少,其余各染料木素劑量組睪丸組織形態(tài)無(wú)明顯病理學(xué)改變。性激素檢測(cè)表明,染毒結(jié)束時(shí)染料木素5mg/kg和50mg/kg劑量組的血清雌二醇水平較正常對(duì)照組比降低(p㩳0.05),染料木素50mg/kg劑量組的血清卵泡刺激素和睪酮水平升高(p㩳0.05);染料木素2000mg/kg劑量組在染毒結(jié)束時(shí)未觀察到激素水平的改變,但到達(dá)成年期時(shí)其雌二醇濃度升高,睪酮濃度降低(p㩳0.05)。精液質(zhì)量分析發(fā)現(xiàn),染料木素2000mg/kg劑量組大鼠在成年期出現(xiàn)精子密度和精子總數(shù)的降低(p㩳0.05)。此外,染毒結(jié)束時(shí)染料木素各劑量組的睪丸erβ表達(dá)未出現(xiàn)明顯改變,但成年期大鼠睪丸β-雌激素受體(estrogenreceptor-β,erβ)的表達(dá)水平隨著染料木素劑量的增高而增高。結(jié)論人群結(jié)果提示,正常膳食攝入大豆異黃酮對(duì)男性精液質(zhì)量沒(méi)有明顯影響,但是對(duì)血清性激素水平會(huì)有一定影響,如降低血清睪酮、黃體生成素、孕酮。動(dòng)物實(shí)驗(yàn)結(jié)果表明,一定水平的染料木素(染料木素5mg/kg組、50mg/kg組、500mg/kg劑量組)對(duì)雄性生殖健康無(wú)明顯不良影響,但是當(dāng)染料木素劑量達(dá)到2000mg/kg時(shí),血清睪酮水平、精子密度、精子總數(shù)下降及精子畸形率增加。
[Abstract]:Objective To study the epidemiological investigation and laboratory test of adult male in Chongqing, and to know the exposure level of soybean isoflavone in adult male in the region, and to determine the correlation between the exposure level of soybean isoflavone and the level of male semen and sex hormone. The effect of soybean isoflavone metabolite on the level of serum sex hormone and the quality of semen was studied by animal experiment. In order to study the negative effects of soybean isoflavone on the reproductive health of male, a reference is made to the effect of soybean isoflavone on the reproductive health of male. Method 1. Population study: The study object was the general adult male recruited in the community of Fengdu county in Chongqing in 2015. The basic information of the study object was collected by the compound questionnaire, and the blood, urine and semen samples of the volunteers were collected on site. The semen parameters were analyzed by high performance liquid chromatography (HPLC), and the semen parameters were analyzed by computer-aided sperm analysis system (CASA), and the sperm morphology and chromatin structure assay were detected by Dff-Quick kit staining. SCSA was used to detect the DNA fragment index of the sperm nucleus. The copy number of the mitochondrial DNA (mtDNA) and the copy number of the mitochondrial DNA (mtDNA) were determined by real-time fluorescence quantitative PCR. The level of serum sex hormone (testosterone, follicle-stimulating hormone, luteinizing hormone, estradiol, progesterone, Prolactin). The relationship between the level of the metabolite of soybean isoflavone and the level of semen and sex hormone was analyzed by using the SPSS16. 0 software. The inclusion criteria of the study object are: (1) the number of abstinence is 2-7 days; (2) male with 18-55 years of age; (3) no history of urinary system infection, history of epididymitis, history of testicular injury, history of varicocele treatment, incomplete testicular decline; (4) no throat and pubic hair, There were no five semen parameters, and (6) the concentration of creatinine (CR) was 0.3 ~ 3g/ L. A total of 161 cases of SD male rats were randomly divided into 6 groups, the normal control group, the genistein 5mg/ kg, the 50mg/ kg, the 500 mg/ kg and the 2000mg/ kg group, and the positive control diethylstilbestrol was 0.1mg/ kg, respectively. Gavage was given for 29 consecutive days. Materials were obtained at the end of the exposure (8-week-old) and after adult (11-week-old), including the organ coefficient, the structure of the testis, the level of sex hormone and the quality of the semen. The expression level of the testis of the testis was detected by Western blot and wb. Results 1. The study of the population: this study examined the level of the metabolite of the human urine genistein, the soybean element, the soybean emodin, the equol and the dihydro-soybean. The results of the jonckheere-terpstra test showed that with the increase of the total soybean isoflavone concentration, the semen volume and the sperm density showed a decreasing trend, and the results were of statistical significance (p = 0.044 and 0.018), and the semen volume increased with the increase of the level of the estriol (p = 0.034). In the sex hormone level, the genistein and the luteinizing hormone (p = 0.019), the soybean emodin and the testosterone (p-0.001), the dihydro-soybean element and the progesterone (p = 0.021) were negatively correlated. After correcting the potential confounders (age, body mass index, abstinence number, smoking status and drinking state) by the multiple linear regression method, the relationship between the level of the soybean isoflavone and the above-mentioned semen parameters and the level of sex hormone still existed. There was no correlation between the soybean isoflavone and the dna damage index (dna fragment index, mtDNA copy number, mtDNA integrity). In addition, the results of multiple linear regression showed that there was an association between the index of sperm dna damage and the quality of semen. The difference was not statistically significant. The results showed that in the end of the exposure, the number of the cells in the cell-tube cavity of the testis of the genistein 2000mg/ kg group was less than that of the normal control group. There were no obvious pathological changes in the form of the testis. The levels of serum estradiol and testosterone in the 50 mg/ kg and 50 mg/ kg group were lower than that in the normal control group (p <0.05), and the level of the serum follicle-stimulating hormone and testosterone in the dose group of the dye-lignan in the dose group was higher than that in the normal control group (p-0.05). The level of testosterone was not observed at the end of the exposure, but the concentration of estradiol increased and the concentration of testosterone decreased (p-0.05) at the end of the exposure. The semen quality analysis found that the density of the sperm and the total number of the sperm in the dose group of the genistein 2000mg/ kg group were lower (p <0.05). In addition, in the end of the exposure, the expression of the testis-estrogen receptor in each dose group of the genistein did not change significantly, but the expression level of the testosterone-estrogen receptor in the adult rat was increased with the increase of the dose of the dye. Conclusion The results suggest that the normal dietary intake of soybean isoflavone has no significant effect on the quality of male semen, but it may have some effect on the serum sex hormone level, such as the reduction of serum testosterone, luteinizing hormone and progesterone. The results of animal experiments showed that a certain level of genistein (mg/ kg, 50mg/ kg, 500mg/ kg) had no significant adverse effects on the reproductive health of the male, but when the dose of the genistein reached 2000mg/ kg, the serum testosterone level, the sperm density, the total number of sperm and the rate of sperm malformation were increased.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R114
[Abstract]:Objective To study the epidemiological investigation and laboratory test of adult male in Chongqing, and to know the exposure level of soybean isoflavone in adult male in the region, and to determine the correlation between the exposure level of soybean isoflavone and the level of male semen and sex hormone. The effect of soybean isoflavone metabolite on the level of serum sex hormone and the quality of semen was studied by animal experiment. In order to study the negative effects of soybean isoflavone on the reproductive health of male, a reference is made to the effect of soybean isoflavone on the reproductive health of male. Method 1. Population study: The study object was the general adult male recruited in the community of Fengdu county in Chongqing in 2015. The basic information of the study object was collected by the compound questionnaire, and the blood, urine and semen samples of the volunteers were collected on site. The semen parameters were analyzed by high performance liquid chromatography (HPLC), and the semen parameters were analyzed by computer-aided sperm analysis system (CASA), and the sperm morphology and chromatin structure assay were detected by Dff-Quick kit staining. SCSA was used to detect the DNA fragment index of the sperm nucleus. The copy number of the mitochondrial DNA (mtDNA) and the copy number of the mitochondrial DNA (mtDNA) were determined by real-time fluorescence quantitative PCR. The level of serum sex hormone (testosterone, follicle-stimulating hormone, luteinizing hormone, estradiol, progesterone, Prolactin). The relationship between the level of the metabolite of soybean isoflavone and the level of semen and sex hormone was analyzed by using the SPSS16. 0 software. The inclusion criteria of the study object are: (1) the number of abstinence is 2-7 days; (2) male with 18-55 years of age; (3) no history of urinary system infection, history of epididymitis, history of testicular injury, history of varicocele treatment, incomplete testicular decline; (4) no throat and pubic hair, There were no five semen parameters, and (6) the concentration of creatinine (CR) was 0.3 ~ 3g/ L. A total of 161 cases of SD male rats were randomly divided into 6 groups, the normal control group, the genistein 5mg/ kg, the 50mg/ kg, the 500 mg/ kg and the 2000mg/ kg group, and the positive control diethylstilbestrol was 0.1mg/ kg, respectively. Gavage was given for 29 consecutive days. Materials were obtained at the end of the exposure (8-week-old) and after adult (11-week-old), including the organ coefficient, the structure of the testis, the level of sex hormone and the quality of the semen. The expression level of the testis of the testis was detected by Western blot and wb. Results 1. The study of the population: this study examined the level of the metabolite of the human urine genistein, the soybean element, the soybean emodin, the equol and the dihydro-soybean. The results of the jonckheere-terpstra test showed that with the increase of the total soybean isoflavone concentration, the semen volume and the sperm density showed a decreasing trend, and the results were of statistical significance (p = 0.044 and 0.018), and the semen volume increased with the increase of the level of the estriol (p = 0.034). In the sex hormone level, the genistein and the luteinizing hormone (p = 0.019), the soybean emodin and the testosterone (p-0.001), the dihydro-soybean element and the progesterone (p = 0.021) were negatively correlated. After correcting the potential confounders (age, body mass index, abstinence number, smoking status and drinking state) by the multiple linear regression method, the relationship between the level of the soybean isoflavone and the above-mentioned semen parameters and the level of sex hormone still existed. There was no correlation between the soybean isoflavone and the dna damage index (dna fragment index, mtDNA copy number, mtDNA integrity). In addition, the results of multiple linear regression showed that there was an association between the index of sperm dna damage and the quality of semen. The difference was not statistically significant. The results showed that in the end of the exposure, the number of the cells in the cell-tube cavity of the testis of the genistein 2000mg/ kg group was less than that of the normal control group. There were no obvious pathological changes in the form of the testis. The levels of serum estradiol and testosterone in the 50 mg/ kg and 50 mg/ kg group were lower than that in the normal control group (p <0.05), and the level of the serum follicle-stimulating hormone and testosterone in the dose group of the dye-lignan in the dose group was higher than that in the normal control group (p-0.05). The level of testosterone was not observed at the end of the exposure, but the concentration of estradiol increased and the concentration of testosterone decreased (p-0.05) at the end of the exposure. The semen quality analysis found that the density of the sperm and the total number of the sperm in the dose group of the genistein 2000mg/ kg group were lower (p <0.05). In addition, in the end of the exposure, the expression of the testis-estrogen receptor in each dose group of the genistein did not change significantly, but the expression level of the testosterone-estrogen receptor in the adult rat was increased with the increase of the dose of the dye. Conclusion The results suggest that the normal dietary intake of soybean isoflavone has no significant effect on the quality of male semen, but it may have some effect on the serum sex hormone level, such as the reduction of serum testosterone, luteinizing hormone and progesterone. The results of animal experiments showed that a certain level of genistein (mg/ kg, 50mg/ kg, 500mg/ kg) had no significant adverse effects on the reproductive health of the male, but when the dose of the genistein reached 2000mg/ kg, the serum testosterone level, the sperm density, the total number of sperm and the rate of sperm malformation were increased.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R114
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