环境激素双酚A对成年SD大鼠睾丸内P450scc和3β-HSD蛋白表达的影响
发布时间：2019-04-30 11:38
【摘要】：目的：当前环境污染越来越重,环境对人类的伤害越来越厉害,尤其是大量化工材料的生产和使用使得环境污染以从未有过的高度被人类重视起来，因为这已经深深危害到人类的健康甚至危害人类下一代，在大量的环境污染物质中环境激素类物质尤其引人关注，其中很重要的一种环境激素类污染物就是双酚A,双酚A存在于日常生活中随处可见的塑料制品中,用途广泛，自然环境中存在半衰期长，难降解，对人类健康影响大。近年来男性的生育能力越来越差，上世纪五六十年代男性精子密度是现在男性精子密度的一倍，这种现象引起科学家的重视，经过大量的研究分析发现环境污染是造成男性生育能力下降的主要因素之一。环境激素双酚A对男性生育能力影响的研究成为了科学家研究的重点。 本实验就是研究环境激素双酚A(bisphenol A,BPA)灌胃染毒雄性SD大鼠后对大鼠睾丸合成雄激素睾酮的影响，并探讨给予大鼠灌胃双酚A后对睾丸间质细胞中睾酮合成关键酶P450scc(胆固醇侧链裂解酶)和3β-HSD(3β-羟基类固醇脱氢酶)蛋白表达的影响,在细胞蛋白水平研究双酚A对雄性大鼠睾酮合成影响的机制,以期待能够探明环境激素双酚A影响雄性生育能力的具体机制,从而能够从根源上预防和解决环境激素双酚A对人类男性生育能力的影响 材料与方法: 1实验动物及材料:实验动物选取清洁级8周龄成年雄性SD(Sprague-Dawley)大鼠,体重在180-200g所有大鼠均分笼饲养于动物房一周后用于实验,每日模拟自然采光12小时,均自由采食全价颗粒饲料,饮用自来水,每天定时染毒,每周染毒5次,染毒8周后用于实验 2实验动物分组及染毒方式：将四十八只大鼠随机分为四组即对照组、低剂量组、中剂量组和高剂量组,每组12只各个实验组分别用玉米油配制成不同剂量浓度的BPA灌胃染毒,低剂量组给予2mg/kgBPA灌胃,中剂量组给予20mg/kgBPA灌胃,高剂量组给予200mg/kgBPA灌胃,对照组每日灌等量玉米油 3实验标本处理：染毒结束后大鼠称重,然后处死心脏取血,静置后离心分离保存血清,应用酶联免疫方法检测血清睾酮的浓度,取睾丸附睾进行称重计算脏器系数,睾丸中性甲醛固定,包埋切片行HE染色和免疫组化染色,对比分析染毒组和对照组差异，结果应用SPSS软件系统进行分析。 结果： 1大鼠日常活动变化：高剂量组染毒大鼠表现出如被毛杂乱、活动减少和软便等中毒症状,同时体重增长缓慢。其余各组大鼠状态正常未见明显异常活动。 2双酚A染毒后体重及睾丸变化：随着BPA染毒剂量的增加，，大鼠体重、睾丸脏器系数逐渐降低，染毒中、高剂量组与对照组相比各脏器指标均有统计学意义(P＜0.05)，且大鼠体重、睾丸脏器系数与BPA染毒剂量相关，染毒剂量越大增长越缓慢。 3大鼠血清睾酮的含量差别:低剂量组大鼠血清睾酮含量略高于对照组,但差异没有统计学意义；中、高剂量染毒组睾酮含量低于对照组,随染毒剂量增加而降低,差异有统计学意义 4睾丸组织结构变化:对照组大鼠睾丸组织结构显示睾丸发育良好,精细小管排列致密,精原细胞、初级精母细胞、次级精母细胞和精子细胞依次排列整齐,大量精子存在管腔内中、高剂量组睾丸发育差精细小管排列松散,间质细胞稀疏,部分精细小管腔内生殖细胞排列紊乱低剂量组大鼠切片未见明显异常 5P450scc在大鼠睾丸间质细胞上的表达:对照组与低剂量组的阳性间质细胞数较多,细胞着色深,吸光度接近，差异没有统计学意义；中、高剂量组P450scc的阳性细胞表达弱,染色浅,吸光度变小，差异有统计学意义 63βHSD在大鼠睾丸间质细胞上的表达:3βHSD在中高剂量组与对照组相比阳性细胞数目减少,染色颜色变浅，吸光度变小，差异有统计学意义在对照组和低剂量染毒组之间没有明显差异，阳性细胞多、染色深 结论：经口摄入环境激素双酚A染毒雄性大鼠可以对雄性大鼠生殖系统产生毒害作用,摄入双酚A达到一定浓度可以影响大鼠体重和睾丸重量变化,较高剂量会抑制睾丸产生睾酮,作用机制可能与抑制睾酮合成关键酶P450scc和3βHSD表达有关
[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
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R114
本文编号：2468740
[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
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R114
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