本文選題：青春期大鼠 + 鄰苯二甲酸酯類��； 參考：《汕頭大學》2008年碩士論文

【摘要】： 背景: 近年來,環(huán)境因素對男性生殖健康的影響越來越引起學者們的注意。鄰苯二甲酸酯類是常用的塑料制品增塑劑,有很強的擬雌激素樣作用,是公認的環(huán)境內(nèi)分泌干擾物,不但可以干擾下丘腦-垂體-睪丸軸的平衡,而且還可以直接或(和)間接地損害精子的生成。青春期是男性生長發(fā)育的關(guān)鍵時期,在此期受到外界環(huán)境不良干擾可能會影響男性生殖功能和個體的生長發(fā)育。最近,有學者認為,我們所處的環(huán)境中含有多種鄰苯二甲酸酯類物質(zhì),所以我們有必要評估兩種或多種鄰苯二甲酸酯類混合物對人類健康的影響。鄰苯二甲酸二(2-乙基己基)酯和鄰苯二甲酸二丁酯是世界上最常用的兩種增塑劑,并且人群最常見的接觸方式是經(jīng)口接觸污染的水或食物。因此,本研究試圖探討鄰苯二甲酸二(2-乙基己基)酯和鄰苯二甲酸二丁酯聯(lián)合經(jīng)口染毒對青春期大鼠生殖內(nèi)分泌功能的影響及其機制;這些研究對阻止、延緩或減弱上述因素對人類生殖健康的損傷有重要的意義。 目的: 1)研究鄰苯二甲酸酯類對雄性性行為的影響及其機制。 2)探討鄰苯二甲酸酯類對睪丸、附睪組織形態(tài)及生精功能的影響,并闡明生殖內(nèi)分泌功能的改變在其中的作用。 3)研究睪丸內(nèi)與精子功能相關(guān)的酶類活性的改變以及睪丸內(nèi)氧化-抗氧化系統(tǒng)的改變,進一步探討這些改變在生殖功能損傷機制中的作用。 4)研究睪丸中銅、鋅含量及銅/鋅比值的變化在鄰苯二甲酸酯類引起生殖損傷中的作用。 材料與方法: 健康雄性SD大鼠(6周齡)56只,分籠飼養(yǎng)于標準動物房(22℃),自由飲食,給予規(guī)律光照(12 h:12 h)。20只健康成年雌性SD大鼠經(jīng)處理后與雄鼠交配,觀察雄鼠性行為學改變。將所有雄鼠隨機分為4組,每組14只,將DEHP和DBP等質(zhì)量溶解于玉米油中,灌胃劑量分別為0(對照組)、375、750、1500 mg/(kg.d)。于染毒2周末各組斷頭法處死7只大鼠。染毒第22~24天觀察雄鼠性行為學的改變,染毒4周末各組斷頭法處死7只大鼠。每天同一時間稱重,灌胃量為5ml/Kg體重。大鼠處死后收集標本,采用組織學方法分析各組睪丸曲細精管直徑大小和生精上皮高度變化,流式細胞分析儀檢測各組睪丸生精細胞的變化,并用放免法檢測血清中睪酮和雌二醇含量的變化;采用紫外分光光度計法檢測血清和睪丸勻漿液中ACP、SDH、SOD和MDA的含量變化以及染毒4周血清中NO的變化;采用石墨爐原子發(fā)光法檢測睪丸組織中銅、鋅含量以及銅/鋅比值的變化。 結(jié)果: 1.大鼠性行為學觀察:在30min內(nèi),對照組和750 mg/kg.d組分別有6只大鼠完成射精,而375 mg/kg.d組和1500 mg/kg.d組分別有7只和5只大鼠完成射精。插入潛伏期在1500mg/kg.d組較對照組和375 mg/kg.d組延長(P 0.05);射精前插入次數(shù)在1500 mg/kg.d組與對照組相比差別有統(tǒng)計學意義(P 0.05)。 2.睪丸組織形態(tài)學改變:曲細精管直徑在染毒2周750 mg/kg.d組和1500 mg/kg.d組較對照組降低,染毒4周各劑量組曲細精管直徑均較對照組明顯減小。染毒2、4周,生精上皮高度在375 mg/kg.d組較對照組降低(P 0.05),在750 mg/kg.d組和1500 mg/kg.d組與對照組相比降低均具有高度統(tǒng)計學意義(P 0.01)。 3.流式細胞儀檢測生精細胞的變化:染毒2、4周,亞單倍體細胞比例在750 mg/kg.d組和1500 mg/kg.d組升高;隨染毒劑量增加,1C細胞和4C比例降低,2C細胞比例升高, G0/G1期細胞比例升高,G2/M期細胞比例降低。 4.血清睪酮(T)、雌二醇(E2)和NO的變化:染毒2周,血清T含量在750 mg/kg.d組和1500 mg/kg.d組較對照組明顯降低(P 0.01);血清中E2在750 mg/kg.d組和1500 mg/kg.d組較對照組明顯升高(P 0.05);染毒4周,血清T和E2含量在750 mg/kg.d組和1500 mg/kg.d組與對照組相比差別均具有高度統(tǒng)計學意義(P 0.01);血清中NO含量在1500 mg/kg.d組較對照組升高(P 0.05)。 5.血清及睪丸勻漿中ACP、SDH、SOD和MDA活性的變化:染毒2周,血清SOD活性在1500 mg/kg.d組明顯比對照組降低(P 0.05),睪丸勻漿中ACP、SOD和MDA在1500 mg/kg.d組與對照組相比差別有統(tǒng)計學意義(P 0.05);染毒4周,血清ACP活性在750 mg/kg.d組與對照組相比有統(tǒng)計學意義(P 0.05),在1500 mg/kg.d組與對照組相比有高度統(tǒng)計學意義(P 0.01),血清SDH活性在1500 mg/kg.d組與對照比差別有統(tǒng)計學意義(P 0.05)。睪丸勻漿中ACP、SDH、SOD和MDA在750 mg/kg.d組與對照組相比差別有統(tǒng)計學意義(P 0.05),在1500 mg/kg.d組各項指標與對照組相比差別有高度統(tǒng)計學意義(P 0.01)。6.睪丸組織中銅、鋅及銅/鋅比值的變化:染毒2周,睪丸組織中銅含量在1500 mg/kg.d組升高(P 0.05),鋅的含量和銅/鋅比值較對照組差別明顯(P 0.05);染毒4周,在1500 mg/kg.d組睪丸組織中鋅含量明顯降低(P 0.05),銅/鋅比值明顯升高(P 0.05)。 結(jié)論: 1.鄰苯二甲酸酯類可以引起雄性大鼠性行為的異常,其機制可能與體內(nèi)性激素水平紊亂和NO水平變化有關(guān)。 2.鄰苯二甲酸酯類對睪丸組織形態(tài)結(jié)構(gòu)有嚴重影響,破壞睪丸生精上皮結(jié)構(gòu)的完整性,干擾生精過程。 3.生精相關(guān)酶活性的改變和睪丸內(nèi)氧化抗氧化系統(tǒng)失衡可能在鄰苯二甲酸酯類引起生精功能障礙的過程中發(fā)揮了重要的作用,并且可能解釋鄰苯二甲酸酯類引起睪丸損傷的某些機制。 4.睪丸內(nèi)銅、鋅含量和銅/鋅比值的變化可能與鄰苯二甲酸酯類引起睪丸損傷有密切關(guān)系。
[Abstract]:Background:
In recent years, the effects of environmental factors on male reproductive health have attracted more and more attention from scholars. Phthalic acid esters are commonly used plastic plasticizers. They have strong estrogen like effects. It is a recognized environmental endocrine disruptor. It can not only interfere with the balance of the hypothalamus pituitary - testicular axis, but also direct or (and). Earthing damage sperm production. Puberty is a critical period for male growth and development. Adverse environmental disturbance may affect male reproductive function and individual growth and development during this period. Recently, some scholars believe that we are in the environment containing a variety of phthalate two formates, so we need to assess two or more kinds. The effects of phthalic acid esters on human health. Phthalic acid two (2- ethyl hexyl) ester and dibutyl phthalate are the two most commonly used plasticizers in the world, and the most common contact mode in the population is water or food contaminated by oral contact. Therefore, this study attempts to explore the two (2- ethyl hexyl) of phthalic acid two formate. The effects and mechanisms of ester and dibutyl phthalate on reproductive endocrine function of puberty rats, which are important to prevent, delay or weaken the damage of human reproductive health.
Objective:
1) to study the effects of o-benzoic acid two esters on male sexual behavior and its mechanism.
2) to explore the effects of o-phthalic acid two esters on testis, epididymis morphology and spermatogenesis, and elucidate the role of reproductive endocrine function in the process.
3) study the changes in the activity of the enzymes related to the spermatozoon function in the testis and the changes in the oxidation antioxidant system in the testis, and further explore the role of these changes in the mechanism of reproductive function damage.
4) to study the effects of copper, zinc content and copper / zinc ratio on the reproductive damage induced by o-benzoic acid two esters.
Materials and methods:
56 healthy male SD rats (6 weeks old) were kept in a standard animal room (22 degrees C), free diet, and regular light (12 h:12 h) were given to male adult female SD rats to mate with male rats after treatment. The male rats were divided into 4 groups randomly, 14 rats in each group were dissolved in corn oil, and the quality of DEHP and DBP was dissolved in corn oil and irrigation. The dosage of the stomach was 0 (control group) and 3757501500 mg/ (kg.d). 7 rats were killed at the end of the 2 weekend. The sex behavior changes of male rats were observed on day 22~24, and 7 rats were killed at the end of the 4 weekend. The weight of the rats was weighed at the same time every day and the amount of gavage was 5ml/Kg. The specimens were collected and the histology method was used after the rats were killed. The changes in testicular spermatogenic cells were detected by flow cytometry, and the changes in serum testosterone and estradiol were detected by flow cytometry. The changes in the content of ACP, SDH, SOD and MDA in serum and testicular homogenate were detected by ultraviolet spectrophotometer. The changes of NO in serum were observed for 4 weeks. The changes of copper, zinc and copper / zinc ratio in testicular tissue were detected by graphite furnace atomic fluorescence spectrometry.
Result:
1. rat sexology observation: in 30min, 6 rats in the control group and the 750 mg/kg.d group had ejaculation, while 7 and 5 rats in the 375 mg/kg.d group and 1500 mg/kg.d group completed ejaculation. The insertion latency was prolonged in the 1500mg/kg.d group compared with the control group and the 375 mg/kg.d group (P 0.05), and the number of pre ejaculatory insertion was in the 1500 mg/kg.d group and the control group. The difference was statistically significant (P 0.05).
2. the morphological changes of the testis: the diameter of the convoluted tubule was lower in the 750 mg/kg.d group and the 1500 mg/kg.d group than the control group. The diameter of the seminiferous tubule of the group was significantly lower than the control group for 4 weeks. The height of the spermatogenic epithelium was lower in the 375 mg/kg.d group than the control group (P 0.05), and in the 750 mg/kg.d group and the 1500 mg/kg.d group and the control group. The decrease was statistically significant (P 0.01).
3. flow cytometry was used to detect the changes in spermatogenic cells: the proportion of the haploid cells in the 750 mg/kg.d group and the 1500 mg/kg.d group was increased in 2,4 weeks. With the increase of the dose, the proportion of 1C cells and 4C decreased, the proportion of 2C cells increased, the proportion of G0/G1 phase cells increased, and the proportion of the cells in G2/M phase decreased.
4. the changes of serum testosterone (T), estradiol (E2) and NO: the content of serum T decreased significantly in the 750 mg/kg.d and 1500 mg/kg.d groups (P 0.01), and the serum E2 in the 750 mg/kg.d group and 1500 mg/kg.d group was significantly higher than the control group (P 0.05), and for 4 weeks, the serum T and 1500 concentrations were compared with the control group and the 750 group and the control group. The difference was statistically significant (P 0.01), and the serum NO level was higher in the 1500 mg/kg.d group than in the control group (P 0.05).
5. changes in the activity of ACP, SDH, SOD and MDA in serum and testicular homogenate: the activity of serum SOD in the 1500 mg/kg.d group was significantly lower than that in the control group (P 0.05), ACP in the homogenate of the testis, SOD and MDA in the 1500 mg/kg.d group was significantly different from the control group (P 0.05). The activity of serum albumin in the 750 group was compared with the control group for 4 weeks. Statistical significance (P 0.05), in the 1500 mg/kg.d group, compared with the control group, there was a significant statistical significance (P 0.01), and the serum SDH activity in the 1500 mg/kg.d group was significantly different from the control ratio (P 0.05). The comparison of ACP, SDH, SOD and MDA in the 750 mg/kg.d group in the testicular homogenate was statistically significant (P 0.05), in the 1500 group of fingers. Compared with the control group, there was a significant difference in statistical significance (P 0.01) the changes in copper, zinc and copper / zinc ratio in.6. testis tissue: the copper content in the testicular tissue was increased in the 1500 mg/kg.d group (P 0.05), the content of zinc and the ratio of copper / zinc were significantly different from those of the control group (P 0.05), and the zinc content in the 1500 mg/kg.d group was significantly higher than that of the control group for 4 weeks. The ratio of copper / zinc was significantly increased (P 0.05) and the ratio of copper / zinc was significantly increased (P 0.05).
Conclusion:
1. o-phthalate two formic acid esters can induce abnormal sexual behavior in male rats, which may be related to the disturbance of sex hormone levels and the change of NO level.
2. o-phthalic acid two formic acid has a serious effect on the morphology and structure of testicular tissue, which destroys the integrity of testicular spermatogenic epithelium and interferes with spermatogenesis.
The changes in the activity of 3. spermatogonia related enzymes and the imbalance of the oxidation antioxidant system in the testis may play an important role in the process of spermatogenic dysfunction induced by phthalate esters, and may explain some mechanisms of testis damage caused by phthalate esters.
4. the change of copper, zinc content and copper / zinc ratio in testes may be closely related to the damage of testicular injury caused by o-benzoic acid two formic acid ester.
【學位授予單位】：汕頭大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R363

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