本文選題：乙烯利 + 生精細(xì)胞　； 參考：《新鄉(xiāng)醫(yī)學(xué)院》2016年碩士論文

【摘要】：背景近半個(gè)世紀(jì)來,不孕不育已經(jīng)成為一個(gè)十分嚴(yán)重的社會(huì)問題,男性不育在所有不孕不育中高達(dá)50%,但是大部分不育患者原因不明,其中環(huán)境因素被認(rèn)為是引起男性不育主要原因,并引發(fā)社會(huì)學(xué)者的極大關(guān)注。乙烯利是應(yīng)用最為廣泛的植物生長(zhǎng)調(diào)節(jié)劑之一,可促進(jìn)植物開花、果實(shí)成熟,主要用于提高植物產(chǎn)量和采摘后果實(shí)的催熟。在生產(chǎn)過程中存在大劑量違規(guī)應(yīng)用現(xiàn)象,研究發(fā)現(xiàn)可在食物中大量殘留。國(guó)外學(xué)者研究發(fā)現(xiàn)乙烯利可使蛋白磷酸化,引起細(xì)胞生物活性改變,國(guó)內(nèi)學(xué)者發(fā)現(xiàn)乙烯利對(duì)生物的生殖系統(tǒng)、內(nèi)分泌系統(tǒng)、免疫系統(tǒng)、神經(jīng)系統(tǒng)均有影響,但目前未見對(duì)生物青春期雄性生殖系統(tǒng)影響的研究報(bào)道,因此本研究通對(duì)青春期雄性SD大鼠采用不同劑量的乙烯利并應(yīng)用不同時(shí)間,觀察體質(zhì)量、睪丸細(xì)胞凋亡及性激素水平變化,探討乙烯利對(duì)雄性大鼠的生殖毒性及性激素水平的影響,為青少年生長(zhǎng)發(fā)育及青壯年的不育提供一定理論基礎(chǔ)。目的建立接觸不同劑量乙烯利的SD大鼠模型,觀察大鼠一般生活狀態(tài)、體重、血清性激素水平、大鼠睪丸組織結(jié)構(gòu)的變化及生精細(xì)胞凋亡變化情況,探討乙烯利對(duì)其生殖毒性及性激素水平的影響,以及對(duì)人體的安全性提供科學(xué)的理論依據(jù),為青少年生長(zhǎng)發(fā)育及青壯年的不育提供一定理論基礎(chǔ)。方法隨機(jī)取Sprague-Dawley(SD)雄性大鼠96只隨機(jī)分為四組:高劑量組、中劑量組、低劑量組、對(duì)照組,每組分別為24只。各實(shí)驗(yàn)組分別給予乙烯利2000 mg/kg、1000 mg/kg、500 mg/kg[約為乙烯利半數(shù)致死量(LD50)的1/2、1/4、1/8],對(duì)照組給予等量生理鹽水。采用經(jīng)口灌胃方法,連續(xù)灌胃,1次/d,每3d稱重1次,根據(jù)體質(zhì)量的變化調(diào)整給藥量。每組分別在7d、14d、21d及停用乙烯利30天時(shí)處死6只。各組處死時(shí)測(cè)體質(zhì)量后心臟穿刺抽血3ml,采血離心后血清置-20℃冰箱備測(cè),將去掉被膜的睪丸稱重后,經(jīng)10%福爾馬林固定后,根據(jù)常規(guī)方法制作成石蠟切片。采用化學(xué)發(fā)光免疫法(chemiluminescence immunoassay,clia)、測(cè)血清雌二醇(estrodiol,E_2)、睪酮(testosterone,t)水平,睪丸切片經(jīng)蘇木素-伊紅染色后光鏡下觀察:平均曲細(xì)精管直徑(meantubulardiametar,mtd)。采用dna聚合酶和末端脫氧核糖核酸轉(zhuǎn)移酶標(biāo)記dna鏈斷端的核酸(tdt-mediateddutpnick-endlabelingtunel)技術(shù)檢測(cè)睪丸生殖細(xì)胞凋亡,高倍鏡計(jì)算凋亡指數(shù)(apoptosisindex,ai)。所有數(shù)值均用平均數(shù)±標(biāo)準(zhǔn)差(喁±s)表示,使用spss20.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析,每一組間數(shù)值比較用單因素方差分析(anova),以p0.05作為顯著性檢驗(yàn)標(biāo)準(zhǔn)。結(jié)果(同一時(shí)間點(diǎn))1各實(shí)驗(yàn)組體質(zhì)量、睪丸質(zhì)量、mtd平均值、生殖細(xì)胞凋亡指數(shù)較對(duì)照組降低(p0.05),其中高劑量組、中劑量組較對(duì)照組顯著降低(p0.01);高劑量組較中劑量組相比差異無統(tǒng)計(jì)學(xué)意義(p0.05)。2在7天和14天時(shí),高劑量組血清E_2水平與中劑量組、低劑量組、對(duì)照組相比較降低,具有統(tǒng)計(jì)學(xué)意義(p0.05);21天時(shí)高劑量組、中劑量組血清E_2水平與低劑量、對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(p0.05);低劑量組與對(duì)照組相比,大鼠生殖血清E_2無統(tǒng)計(jì)學(xué)意義(p0.05)。3高劑量組、中劑量組血清t水平與低劑量組、對(duì)照組相比較降低,具有統(tǒng)計(jì)學(xué)意(p0.05);高劑量組較低劑量、對(duì)照組相比具有顯著差異,差異有統(tǒng)計(jì)學(xué)意義(p0.01);低劑量組與對(duì)照組相比無統(tǒng)計(jì)學(xué)意義(p0.05)。(同一劑量組)1與對(duì)照組相比,大鼠體質(zhì)量、睪丸質(zhì)量、mtd平均值隨著乙烯利接觸時(shí)間的延長(zhǎng)而下降,生殖細(xì)胞凋亡指數(shù)隨時(shí)間的延長(zhǎng)而增加;單因素方差分析顯示,乙烯利對(duì)大鼠體質(zhì)量、睪丸質(zhì)量增長(zhǎng)、mtd平均值增長(zhǎng)、生精細(xì)胞凋亡率有影響(P0.05);2高劑量組、中劑量組大鼠血清E2、T水平隨時(shí)間的延長(zhǎng),逐漸增高,但趨勢(shì)較平穩(wěn),單因素方差分析顯示,一定劑量乙烯利對(duì)血清E2和T水平有一定影響(P0.05)。(停止應(yīng)用乙烯利30天后)各實(shí)驗(yàn)組較對(duì)照組體質(zhì)量、睪丸質(zhì)量、睪丸MTD平均值、生殖細(xì)胞凋亡指數(shù)、血清E2和T水平無顯著差別,逐漸恢復(fù)至接近正常水平(P0.05)。結(jié)論乙烯利對(duì)大鼠有一定的生殖毒性作用,可引起睪丸組織學(xué)結(jié)構(gòu)改變、性激素水平降低,增加生精細(xì)胞凋亡,導(dǎo)致生精能力降低,這可能是導(dǎo)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.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R698.2

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5 張海江;IL-23/IL-17通路介導(dǎo)SD大鼠視網(wǎng)膜缺血再灌注損傷的作用機(jī)制及姜黃素的干預(yù)研究[D];武漢大學(xué);2015年

6 周軍;介入硬化劑聚桂醇治療SD大鼠子宮內(nèi)膜異位囊腫的實(shí)驗(yàn)研究[D];武漢大學(xué);2014年

7 張姍姍;姜黃素對(duì)口腔黏膜下纖維性變SD大鼠模型抗纖維化作用及機(jī)制的研究[D];中南大學(xué);2012年

8 馬志飛;“中低溫低流量”腦損傷SD大鼠模型的制作以及缺血預(yù)適應(yīng)腦保護(hù)作用機(jī)制的研究[D];南京醫(yī)科大學(xué);2014年

9 陳會(huì)強(qiáng);SD大鼠主動(dòng)脈瓣間質(zhì)細(xì)胞自發(fā)鈣化模型的建立及阿托伐他汀和氨氯地平的影響[D];河北醫(yī)科大學(xué);2012年

10 鹿楠;SD大鼠腰2移位修復(fù)骶叢撕脫傷的實(shí)驗(yàn)研究[D];第二軍醫(yī)大學(xué);2013年

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2 李翠;外源性內(nèi)脂素對(duì)SD大鼠子宮內(nèi)膜容受性的影響[D];河北聯(lián)合大學(xué);2014年

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