大豆异黄酮对男（雄）性精液质量及性激素水平的影响
发布时间：2018-11-17 11:04
【摘要】：目的通过对重庆地区普通成年男性流行病学调查及实验室检测,了解该地区成年男性大豆异黄酮内暴露水平,明确大豆异黄酮内暴露水平与男性精液质量和性激素水平之间的关联。通过动物实验研究探讨不同剂量下的大豆异黄酮代谢物对血清性激素水平及精液质量的影响。为探讨大豆异黄酮对男性生殖健康的不良影响提供参考依据。方法1、人群研究:研究对象为2015年在重庆市丰都县社区招募的普通成年男性,通过复合式问卷收集研究对象的基本信息,现场采集志愿者的血液、尿液及精液样本。采用高效液相色谱(High performance liquid chromatography,HPLC)分析人群尿液大豆异黄酮代谢物水平,计算机辅助精子分析系统(Computer-aided sperm analyzer,CASA)进行精液参数分析,Diff-Quick试剂盒染色检测精子形态,染色质结构分析法(Sperm chromatin structure assay,SCSA)检测精子核DNA碎片指数,实时荧光定量PCR方法测定精子线粒体DNA(Mitochondrial DNA,mtDNA)拷贝数,长链PCR方法检测mtDNA完整性,化学发光法检测人群血清性激素水平(睾酮、卵泡刺激素、黄体生成素、雌二醇、孕酮、泌乳素)。采用SPSS16.0软件分析了大豆异黄酮尿代谢物水平与精液质量及性激素水平之间的关联。研究对象的纳入标准为:(1)禁欲天数为2~7天;(2)年龄18~55周岁男性;(3)无泌尿系统感染史、附睾炎病史、睾丸损伤疾病史、精索静脉曲张治疗史、睾丸下降不全史;(4)无喉结、阴毛、乳腺发育异常、阴茎异常、睾丸损伤、附睾炎症、精索静脉曲张、隐睾术后;(5)无五项精液参数均缺失者;(6)尿肌酐(Creatinine,CR)浓度在0.3~3g/L。最终纳入161例作为研究对象。2、动物实验研究:采用60只4周龄(青春期)SD雄性大鼠进行亚慢性染毒,随机分为6组,分别是正常对照组、染料木素5mg/kg、50mg/kg、500 mg/kg和2000mg/kg组、阳性对照己烯雌酚0.1mg/kg组,连续灌胃29天。分别在染毒结束时(8周龄)以及动物成年后(11周龄)进行取材,观察指标包括脏器系数、睾丸组织结构、性激素水平及精液质量。采用蛋白质印迹(westernblot,wb)检测睾丸erβ的表达水平。结果1、人群研究:本次研究检测了人群尿液染料木素、大豆苷元,黄豆黄素,雌马酚,二氢大豆苷元代谢物水平。jonckheere-terpstra检验结果显示,随着总的大豆异黄酮浓度等级的增加,精液体积和精子密度呈现递减趋势,结果有统计学意义(p=0.044和0.018),随着雌马酚水平的增加,精液体积呈增加的趋势(p=0.034)。在性激素水平上,染料木素与黄体生成素(p=0.019),黄豆黄素与睾酮(p㩳0.001),二氢大豆苷元与孕酮(p=0.021)均呈负相关。采用多元线性回归方法校正潜在混杂因素(年龄、体质指数、禁欲天数、吸烟状态、饮酒状态)后,大豆异黄酮尿代谢物水平与上述精液参数关联消失,与性激素水平关联依旧存在。本研究未发现大豆异黄酮与dna损伤指标(dna碎片指数,mtdna拷贝数,mtdna完整性)之间存在关联。此外多元线性回归结果显示,精子dna损伤指标与精液质量之间存在关联。2、动物实验研究:大鼠青春期摄入染料木素后,染毒结束时及成年期染料木素各剂量组大鼠体重及脏器系数较正常对照组比,差异无统计学意义。he染色结果显示染毒结束时染料木素2000mg/kg组睾丸组织生精小管细胞管腔中细胞稀疏,排列紊乱,成年期染料木素2000mg/kg组大鼠生精小管管腔内精子数量较正常对照组比减少,其余各染料木素剂量组睾丸组织形态无明显病理学改变。性激素检测表明,染毒结束时染料木素5mg/kg和50mg/kg剂量组的血清雌二醇水平较正常对照组比降低(p㩳0.05),染料木素50mg/kg剂量组的血清卵泡刺激素和睾酮水平升高(p㩳0.05);染料木素2000mg/kg剂量组在染毒结束时未观察到激素水平的改变,但到达成年期时其雌二醇浓度升高,睾酮浓度降低(p㩳0.05)。精液质量分析发现,染料木素2000mg/kg剂量组大鼠在成年期出现精子密度和精子总数的降低(p㩳0.05)。此外,染毒结束时染料木素各剂量组的睾丸erβ表达未出现明显改变,但成年期大鼠睾丸β-雌激素受体(estrogenreceptor-β,erβ)的表达水平随着染料木素剂量的增高而增高。结论人群结果提示,正常膳食摄入大豆异黄酮对男性精液质量没有明显影响,但是对血清性激素水平会有一定影响,如降低血清睾酮、黄体生成素、孕酮。动物实验结果表明,一定水平的染料木素(染料木素5mg/kg组、50mg/kg组、500mg/kg剂量组)对雄性生殖健康无明显不良影响,但是当染料木素剂量达到2000mg/kg时,血清睾酮水平、精子密度、精子总数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.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：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.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R114
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