乙烯利对青春期SD大鼠生精细胞凋亡及性激素水平的影响

发布时间：2018-06-05 23:41

本文选题：乙烯利 + 生精细胞　；参考：《新乡医学院》2016年硕士论文

【摘要】：背景近半个世纪来,不孕不育已经成为一个十分严重的社会问题,男性不育在所有不孕不育中高达50%,但是大部分不育患者原因不明,其中环境因素被认为是引起男性不育主要原因,并引发社会学者的极大关注。乙烯利是应用最为广泛的植物生长调节剂之一,可促进植物开花、果实成熟,主要用于提高植物产量和采摘后果实的催熟。在生产过程中存在大剂量违规应用现象,研究发现可在食物中大量残留。国外学者研究发现乙烯利可使蛋白磷酸化,引起细胞生物活性改变,国内学者发现乙烯利对生物的生殖系统、内分泌系统、免疫系统、神经系统均有影响,但目前未见对生物青春期雄性生殖系统影响的研究报道,因此本研究通对青春期雄性SD大鼠采用不同剂量的乙烯利并应用不同时间,观察体质量、睾丸细胞凋亡及性激素水平变化,探讨乙烯利对雄性大鼠的生殖毒性及性激素水平的影响,为青少年生长发育及青壮年的不育提供一定理论基础。目的建立接触不同剂量乙烯利的SD大鼠模型,观察大鼠一般生活状态、体重、血清性激素水平、大鼠睾丸组织结构的变化及生精细胞凋亡变化情况,探讨乙烯利对其生殖毒性及性激素水平的影响,以及对人体的安全性提供科学的理论依据,为青少年生长发育及青壮年的不育提供一定理论基础。方法随机取Sprague-Dawley(SD)雄性大鼠96只随机分为四组:高剂量组、中剂量组、低剂量组、对照组,每组分别为24只。各实验组分别给予乙烯利2000 mg/kg、1000 mg/kg、500 mg/kg[约为乙烯利半数致死量(LD50)的1/2、1/4、1/8],对照组给予等量生理盐水。采用经口灌胃方法,连续灌胃,1次/d,每3d称重1次,根据体质量的变化调整给药量。每组分别在7d、14d、21d及停用乙烯利30天时处死6只。各组处死时测体质量后心脏穿刺抽血3ml,采血离心后血清置-20℃冰箱备测,将去掉被膜的睾丸称重后,经10%福尔马林固定后,根据常规方法制作成石蜡切片。采用化学发光免疫法(chemiluminescence immunoassay,clia)、测血清雌二醇(estrodiol,E_2)、睾酮(testosterone,t)水平,睾丸切片经苏木素-伊红染色后光镜下观察:平均曲细精管直径(meantubulardiametar,mtd)。采用dna聚合酶和末端脱氧核糖核酸转移酶标记dna链断端的核酸(tdt-mediateddutpnick-endlabelingtunel)技术检测睾丸生殖细胞凋亡,高倍镜计算凋亡指数(apoptosisindex,ai)。所有数值均用平均数±标准差(喁±s)表示,使用spss20.0软件进行统计学分析,每一组间数值比较用单因素方差分析(anova),以p0.05作为显著性检验标准。结果(同一时间点)1各实验组体质量、睾丸质量、mtd平均值、生殖细胞凋亡指数较对照组降低(p0.05),其中高剂量组、中剂量组较对照组显著降低(p0.01);高剂量组较中剂量组相比差异无统计学意义(p0.05)。2在7天和14天时,高剂量组血清E_2水平与中剂量组、低剂量组、对照组相比较降低,具有统计学意义(p0.05);21天时高剂量组、中剂量组血清E_2水平与低剂量、对照组相比差异有统计学意义(p0.05);低剂量组与对照组相比,大鼠生殖血清E_2无统计学意义(p0.05)。3高剂量组、中剂量组血清t水平与低剂量组、对照组相比较降低,具有统计学意(p0.05);高剂量组较低剂量、对照组相比具有显著差异,差异有统计学意义(p0.01);低剂量组与对照组相比无统计学意义(p0.05)。(同一剂量组)1与对照组相比,大鼠体质量、睾丸质量、mtd平均值随着乙烯利接触时间的延长而下降,生殖细胞凋亡指数随时间的延长而增加;单因素方差分析显示,乙烯利对大鼠体质量、睾丸质量增长、mtd平均值增长、生精细胞凋亡率有影响(P0.05);2高剂量组、中剂量组大鼠血清E2、T水平随时间的延长,逐渐增高,但趋势较平稳,单因素方差分析显示,一定剂量乙烯利对血清E2和T水平有一定影响(P0.05)。(停止应用乙烯利30天后)各实验组较对照组体质量、睾丸质量、睾丸MTD平均值、生殖细胞凋亡指数、血清E2和T水平无显著差别,逐渐恢复至接近正常水平(P0.05)。结论乙烯利对大鼠有一定的生殖毒性作用,可引起睾丸组织学结构改变、性激素水平降低,增加生精细胞凋亡,导致生精能力降低,这可能是导致男性不育的主要原因之一。
[Abstract]:Infertility has become a very serious social problem for nearly half a century. Male infertility is up to 50% in all infertility, but most of the infertile patients are unexplained. Environmental factors are considered to be the main causes of male infertility and cause great concern for social scholars. Ethephon is the most widely used. One of the plant growth regulators, which can promote flowering and ripening of plants, mainly used to improve plant yield and ripening of fruit after harvesting. There is a large dose of illegal application in the process of production. The study found that there is a large amount of residue in the food. Domestic scholars have found that ethephon has an effect on the reproductive system, endocrine system, immune system and nervous system of biological organisms. However, there is no report on the effects of Ethephon on the reproductive system of male adolescent male reproductive system. Therefore, this study used different doses of ethephon for adolescent male SD rats and used different time to observe body mass and testosterone. The effect of Ethephon on the reproductive toxicity and sex hormone level of the male rats was discussed in order to provide a theoretical basis for the growth and growth of young adults and the sterility of young adults. Objective to establish a SD rat model with different doses of ethephon, and to observe the general state of life, weight, and serum sex hormone levels in rats. The changes in the structure of the testis and the changes of spermatogenic cell apoptosis in the rats, the effects of Ethephon on its reproductive toxicity and sex hormone levels, and the scientific basis for the safety of the human body were provided to provide a theoretical basis for the growth and development of young adults and the infertility in young adults. Methods Sprague-Dawley (SD) male rats were taken randomly. 96 rats were randomly divided into four groups: high dose group, medium dose group, low dose group, and control group, 24 rats in each group respectively. The experimental groups were given ethephon 2000 mg/kg, 1000 mg/kg, 500 mg/kg[about 1/2,1/4,1/8] of ethephon half lethal dose (LD50), and the control group was given equal amount of biological saline. Through oral gavage, continuous gavage, 1 times /d, each 3D The weight was adjusted to 1 times according to the change of body mass. Each group died of 6 after 30 days of 7D, 14d, 21d and discontinuation of ethephon. Each group died of body mass after cardiac puncture and pumped 3ml. After the centrifugation, the serum was set at -20 centigrade, then the testicle was weighed off, and after 10% formalin was fixed, it was made into stone according to the conventional method. Paraffin slices. Chemiluminescence immunoassay (CLIA) was used to measure serum estradiol (estrodiol, E_2), testosterone (testosterone, t) levels. The testicular slices were observed under the light microscope after hematoxylin eosin staining: the average diameter of the tubules (meantubulardiametar, MTD). Using DNA polymerase and terminal deoxyribonucleic acid transferase The nucleic acid (tdt-mediateddutpnick-endlabelingtunel) technique marked the broken end of the DNA chain was used to detect the apoptosis of testicular germ cells, and the apoptosis index (apoptosisindex, AI) was calculated by high magnification. All the values were expressed with the mean standard deviation (s), and the spss20.0 software was used for statistical analysis. The single factor analysis of variance (ANOVA) was used in each group. P0.05 as a significant test standard. Results (the same time point) 1 experimental group body mass, testicular mass, MTD average value, the reproductive cell apoptosis index lower than the control group (P0.05), the high dose group, middle dose group is significantly lower than the control group (P0.01), compared with the middle dose group, the difference is not statistically significant (P0.05).2 at 7 days and the difference between the high dose group and the middle dose group (P0.05). At 14 days, the level of serum E_2 in the high dose group was significantly lower than that in the middle dose group, the low dose group and the control group (P0.05). At the 21 day high dose group, the serum E_2 level in the middle dose group was statistically significant compared with the low dose group (P0.05), and the low dose group had no statistical significance (P) compared with the control group (P). 0.05).3 high dose group, the level of serum T in the middle dose group was lower than that in the low dose group, with a statistically significant difference (P0.05), the high dose group had a significant difference compared with the control group, and the difference was statistically significant (P0.01); the low dose group was not statistically significant compared with the control group (P0.05). (the same dose group) was larger than the control group. The mass of rats, the quality of the testis, the MTD average decreased with the prolongation of the ethephon contact time, and the apoptotic index of the germ cells increased with the prolongation of the time. The single factor variance analysis showed that ethephon had an effect on the mass of the rats, the increase of the testicular mass, the average increase of MTD, the rate of fine cell apoptosis (P0.05), and the 2 high dose group, the medium dose group was large. The level of E2 and T in rat serum increased gradually with time, but the trend was more stable. One factor variance analysis showed that a certain dose of ethephon had a certain effect on the level of serum E2 and T (P0.05). (30 days after the use of ethephon), the mass of the experimental group, the quality of the testis, the mean value of MTD, the apoptotic index of the germ cells, the serum E2 and T. There was no significant difference in level, and gradually recovered to the normal level (P0.05). Conclusion ethephon had a certain reproductive toxicity on rats, which could cause the histological structure of the testis, decrease the level of sex hormone, increase the apoptosis of spermatogenic cells and reduce the ability of spermatogenesis, which may be one of the main causes of male infertility.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R698.2

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