發(fā)布時(shí)間：2019-04-30 11:38

【摘要】：目的：當(dāng)前環(huán)境污染越來越重,環(huán)境對人類的傷害越來越厲害,尤其是大量化工材料的生產(chǎn)和使用使得環(huán)境污染以從未有過的高度被人類重視起來，因?yàn)檫@已經(jīng)深深危害到人類的健康甚至危害人類下一代，在大量的環(huán)境污染物質(zhì)中環(huán)境激素類物質(zhì)尤其引人關(guān)注，其中很重要的一種環(huán)境激素類污染物就是雙酚A,雙酚A存在于日常生活中隨處可見的塑料制品中,用途廣泛，自然環(huán)境中存在半衰期長，難降解，對人類健康影響大。近年來男性的生育能力越來越差，上世紀(jì)五六十年代男性精子密度是現(xiàn)在男性精子密度的一倍，這種現(xiàn)象引起科學(xué)家的重視，經(jīng)過大量的研究分析發(fā)現(xiàn)環(huán)境污染是造成男性生育能力下降的主要因素之一。環(huán)境激素雙酚A對男性生育能力影響的研究成為了科學(xué)家研究的重點(diǎn)。 本實(shí)驗(yàn)就是研究環(huán)境激素雙酚A(bisphenol A,BPA)灌胃染毒雄性SD大鼠后對大鼠睪丸合成雄激素睪酮的影響，并探討給予大鼠灌胃雙酚A后對睪丸間質(zhì)細(xì)胞中睪酮合成關(guān)鍵酶P450scc(膽固醇側(cè)鏈裂解酶)和3β-HSD(3β-羥基類固醇脫氫酶)蛋白表達(dá)的影響,在細(xì)胞蛋白水平研究雙酚A對雄性大鼠睪酮合成影響的機(jī)制,以期待能夠探明環(huán)境激素雙酚A影響雄性生育能力的具體機(jī)制,從而能夠從根源上預(yù)防和解決環(huán)境激素雙酚A對人類男性生育能力的影響 材料與方法: 1實(shí)驗(yàn)動物及材料:實(shí)驗(yàn)動物選取清潔級8周齡成年雄性SD(Sprague-Dawley)大鼠,體重在180-200g所有大鼠均分籠飼養(yǎng)于動物房一周后用于實(shí)驗(yàn),每日模擬自然采光12小時(shí),均自由采食全價(jià)顆粒飼料,飲用自來水,每天定時(shí)染毒,每周染毒5次,染毒8周后用于實(shí)驗(yàn) 2實(shí)驗(yàn)動物分組及染毒方式：將四十八只大鼠隨機(jī)分為四組即對照組、低劑量組、中劑量組和高劑量組,每組12只各個(gè)實(shí)驗(yàn)組分別用玉米油配制成不同劑量濃度的BPA灌胃染毒,低劑量組給予2mg/kgBPA灌胃,中劑量組給予20mg/kgBPA灌胃,高劑量組給予200mg/kgBPA灌胃,對照組每日灌等量玉米油 3實(shí)驗(yàn)標(biāo)本處理：染毒結(jié)束后大鼠稱重,然后處死心臟取血,靜置后離心分離保存血清,應(yīng)用酶聯(lián)免疫方法檢測血清睪酮的濃度,取睪丸附睪進(jìn)行稱重計(jì)算臟器系數(shù),睪丸中性甲醛固定,包埋切片行HE染色和免疫組化染色,對比分析染毒組和對照組差異，結(jié)果應(yīng)用SPSS軟件系統(tǒng)進(jìn)行分析。 結(jié)果： 1大鼠日�；顒幼兓焊邉┝拷M染毒大鼠表現(xiàn)出如被毛雜亂、活動減少和軟便等中毒癥狀,同時(shí)體重增長緩慢。其余各組大鼠狀態(tài)正常未見明顯異�；顒�。 2雙酚A染毒后體重及睪丸變化：隨著BPA染毒劑量的增加，，大鼠體重、睪丸臟器系數(shù)逐漸降低，染毒中、高劑量組與對照組相比各臟器指標(biāo)均有統(tǒng)計(jì)學(xué)意義(P＜0.05)，且大鼠體重、睪丸臟器系數(shù)與BPA染毒劑量相關(guān)，染毒劑量越大增長越緩慢。 3大鼠血清睪酮的含量差別:低劑量組大鼠血清睪酮含量略高于對照組,但差異沒有統(tǒng)計(jì)學(xué)意義；中、高劑量染毒組睪酮含量低于對照組,隨染毒劑量增加而降低,差異有統(tǒng)計(jì)學(xué)意義 4睪丸組織結(jié)構(gòu)變化:對照組大鼠睪丸組織結(jié)構(gòu)顯示睪丸發(fā)育良好,精細(xì)小管排列致密,精原細(xì)胞、初級精母細(xì)胞、次級精母細(xì)胞和精子細(xì)胞依次排列整齊,大量精子存在管腔內(nèi)中、高劑量組睪丸發(fā)育差精細(xì)小管排列松散,間質(zhì)細(xì)胞稀疏,部分精細(xì)小管腔內(nèi)生殖細(xì)胞排列紊亂低劑量組大鼠切片未見明顯異常 5P450scc在大鼠睪丸間質(zhì)細(xì)胞上的表達(dá):對照組與低劑量組的陽性間質(zhì)細(xì)胞數(shù)較多,細(xì)胞著色深,吸光度接近，差異沒有統(tǒng)計(jì)學(xué)意義；中、高劑量組P450scc的陽性細(xì)胞表達(dá)弱,染色淺,吸光度變小，差異有統(tǒng)計(jì)學(xué)意義 63βHSD在大鼠睪丸間質(zhì)細(xì)胞上的表達(dá):3βHSD在中高劑量組與對照組相比陽性細(xì)胞數(shù)目減少,染色顏色變淺，吸光度變小，差異有統(tǒng)計(jì)學(xué)意義在對照組和低劑量染毒組之間沒有明顯差異，陽性細(xì)胞多、染色深 結(jié)論：經(jīng)口攝入環(huán)境激素雙酚A染毒雄性大鼠可以對雄性大鼠生殖系統(tǒng)產(chǎn)生毒害作用,攝入雙酚A達(dá)到一定濃度可以影響大鼠體重和睪丸重量變化,較高劑量會抑制睪丸產(chǎn)生睪酮,作用機(jī)制可能與抑制睪酮合成關(guān)鍵酶P450scc和3βHSD表達(dá)有關(guān)
[Abstract]:Objective: At present, the environmental pollution is becoming more and more serious, the harm of the environment to the human being more and more serious, especially the production and use of a large amount of chemical materials, so that the environmental pollution has not been attached to the human being at a high level, Because this has been deeply harmful to the human health and even to the next generation of human beings, environmental pollution-related substances are particularly interesting in a large number of environmental pollution substances, Bisphenol A is widely used in the daily life, and has wide application and long half-life in the natural environment, which is difficult to degrade and has great influence on human health. In recent years, male fertility is becoming more and more poor. In the 1950s and 1960s, male sperm density was one time that of male sperm, which caused the attention of scientists and found that environmental pollution was one of the main factors that caused the decline of male fertility. The study of the effect of environmental hormone bisphenol A on male fertility has become the focus of the research of scientists. This experiment is to study the effect of environmental hormone bisphenol A (BPA) on the synthesis of testosterone in the testis of male SD rats. The effect of bisphenol A on the expression of the key enzyme P450scc (cholesterol side chain cleavage enzyme) and the 3-hydroxy-HSD (3-hydroxy-steroid dehydrogenase) protein was studied and the effect of bisphenol A on the testosterone synthesis in male rats was studied at the level of cellular protein. in ord to be able to identify that specific mechanism of the effect of environmental hormone bisphenol A on male fertility, it is possible to prevent and solve the effect of environmental hormone bisphenol A on human male fertility from the root cause impact material and method:1 experiment The animals and materials: The experimental animals selected the adult male SD (Sprague-Dawley) rats at 8 weeks of age, and the weight was 180-200 g. All rats were fed into the animal room for one week for the experiment. The daily simulated natural lighting for 12 hours was free to feed the complete pellet feed and tap the tap water. 5 times in the week,8 to 8 Methods: Forty-eight rats were randomly divided into four groups: control group, low-dose group, middle-dose group and high-dose group. In the low-dose group,2 mg/ kg of BPA and 20 mg/ kg of BPA were given in the low-dose group, and the high-dose group was given 200 mg/ kg of BPA and the control group. Treatment of three experimental specimens of the same amount of corn oil daily: the rats were weighed after the exposure, and then the heart was sacrificed and the blood was removed and then allowed to stand. The serum was preserved by centrifugal separation, and the serum testosterone concentration was detected by enzyme-linked immunosorbent assay. The testis and epididymis were weighed to calculate the organ coefficient, the testis neutral formaldehyde was fixed, the embedded sections were stained with HE and the immunohistochemical staining, and the difference between the exposure group and the control group was compared. The results were applied to the SPS. S Software The results were as follows:1. The daily activity of the rats: the high-dose group of the rats exposed to the high-dose group, such as the hair, the activity, the soft stool, etc. The symptoms of poisoning, while the weight growth was slow. The rest of the groups were large There was no obvious abnormal activity in the state of rats. Body weight and testis changes after the exposure of bisphenol A: As the dose of BPA was increased, the weight of the rats and the coefficient of the testis organ were gradually decreased, and the high-dose group and the control group had statistical significance in comparison with the control group. (P <0.05), and the body weight, the testis organ coefficient and the BPA-staining agent. The increase of serum testosterone in rats with low-dose group was slightly higher than that in the control group, but the difference was not statistically significant. The amount is lower than that of the control group. In the control group, the testicular tissue structure of the control group showed that the testis was well developed, the fine tubules were arranged dense, the spermatogonia, the primary spermatocyte, the secondary spermatocyte and the sperm were arranged. The cells are orderly arranged, and a large amount of the sperm is in the lumen, and the high-dose group of the testis development difference fine tubules are loose, the interstitial cells are sparse, and the part of the fine small-lumen internal reproduction No significant abnormal 5 P450scc was found in the rat's testis interstitial cells in the low-dose group. Expression: The number of positive mesenchymal cells in the control group and the low-dose group was much higher, the cell color depth, the absorbance was close, the difference was not statistically significant, and the high-dose group P450 positive cells of scc The expression of 63% HSD in the rat's testis interstitial cells was significantly lower than that in the control group. The number of positive cells decreased in the middle and high dose group compared with the control group. The calculated significance was in the control group and the control group. There was no significant difference in the low-dose exposure group. The positive cells were more and the staining was deep. The oral intake of the male rats exposed to the environmental hormone bisphenol A could have a toxic effect on the reproductive system of the male rats. The intake of bisphenol A could have an effect on the concentration of bisphenol A. The weight of the rat and the weight of the testis vary, and the higher dose may inhibit the production of testosterone in the testis, and the mechanism of action may be associated with
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R114

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