鎘暴露對(duì)未成年雄性小鼠的生殖毒性研究
發(fā)布時(shí)間：2018-07-21 13:53
【摘要】：目的探索鎘染毒對(duì)未成年雄性小鼠生殖系統(tǒng)的損傷及生育能力的影響,為進(jìn)一步研究鎘的生殖毒性積累基礎(chǔ)資料。 方法96只四周齡SPF級(jí)雄性NIH小鼠隨機(jī)分為正常對(duì)照組,低、中、高劑量組,每組24只。按照總劑量分別為0、0.84、1.68、3.36mg Cd/kg體重腹腔注射生理鹽水或氯化鎘(CdCl2)和β-巰基乙醇(ME)的混合溶液(二者濃度比為1：20),1次/天,連續(xù)5天。給藥結(jié)束后的第30,60,90,120天,各組分別隨機(jī)取6只動(dòng)物,進(jìn)行如下處理： 1.交配能力檢測(cè)：將雌雄小鼠按1：1合籠,雌鼠放入后,記錄雄鼠首次捕捉雌鼠的時(shí)間即捕捉潛伏期及20min內(nèi)雄鼠捕捉雌鼠的次數(shù)。 2.雄鼠生殖毒性檢測(cè)：①采血,離心取血清,測(cè)定血清睪酮(T)。②解剖取腦、心、肺、肝、脾、腎、睪丸,測(cè)臟器系數(shù)。③留取右腎、右睪丸和部分肝,用原子吸收光譜儀(AAS)測(cè)定其中的鎘含量。④取右側(cè)附睪觀察精子畸形率。⑤固定肝臟、脾臟、左腎、左睪丸和左附睪進(jìn)行病理檢查。 3.雌鼠交配率和受孕率檢測(cè)：將雄鼠與雌鼠按1：2合籠,發(fā)現(xiàn)陰道栓當(dāng)天記為妊娠第0天(GDO),合籠12d后計(jì)算雌鼠交配率,在GD14,確定成功懷孕雌鼠的數(shù)量,計(jì)算受孕率。 4.胎鼠檢測(cè)：每組隨機(jī)取一半受孕雌鼠,于GD17處死取出胎鼠。進(jìn)行一般觀察,并測(cè)量體長(zhǎng)、尾長(zhǎng)及體重。再對(duì)胎鼠分別進(jìn)行骨骼畸形和內(nèi)臟畸形檢查。 5.仔鼠檢測(cè)：讓每組另一半受孕雌鼠自然分娩,仔鼠21日齡時(shí)稱(chēng)重,并觀察性別。 6.窩平均活胎數(shù)統(tǒng)計(jì)：對(duì)各批次各組孕鼠的窩平均活胎數(shù)(包括胎鼠和仔鼠)進(jìn)行統(tǒng)計(jì)。 結(jié)果1.交配能力檢測(cè)結(jié)果：30,60,90,120d批次染鎘雄鼠的交配能力與對(duì)照組相比未出現(xiàn)顯著變化。 2.雄鼠生殖毒性檢測(cè)結(jié)果：①血清T檢測(cè)結(jié)果：30d,60d,120d批次的1.68、3.36mg Cd/kg組雄鼠的血清T含量顯著低于正常對(duì)照(P0.05),90d批次的所有染鎘雄鼠血清T含量均顯著低于正常對(duì)照組(P0.05)。②120d批次的0.84mg Cd/kg組肝臟、肺臟,1.68mg Cd/kg組肝臟的臟器系數(shù)較正常對(duì)照組顯著升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。③臟器鎘含量測(cè)定結(jié)果：各批次染鎘雄鼠肝、腎、睪丸鎘含量均顯著高于正常對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。④精子畸形率：各批次染鎘雄鼠精子畸形率與正常對(duì)照組相比,均顯著升高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。⑤臟器病理檢查結(jié)果：染鎘動(dòng)物腎臟病理變化主要以間質(zhì)炎細(xì)胞浸潤(rùn),腎小管上皮細(xì)胞變性,慢性腎盂腎炎為主。肝臟的病理改變主要有：局灶性炎細(xì)胞浸潤(rùn),肝臟髓外造血,肝細(xì)胞核腫脹等。脾臟也出現(xiàn)了髓外造血。睪丸的病理改變主要以部分生精小管萎縮,部分動(dòng)物睪丸組織壞死為主。附睪的病理改變主要有：附睪管內(nèi)精子消失,部分附睪管內(nèi)可見(jiàn)脫落生精細(xì)胞。 3.雌鼠交配率和受孕率檢測(cè)結(jié)果：各批次染鎘組雌鼠的交配率和受孕率與對(duì)照組相比均未出現(xiàn)顯著變化。 4.胎鼠檢測(cè)結(jié)果：各批次染鎘組胎鼠的體長(zhǎng)、尾長(zhǎng)、體重與正常對(duì)照組相比未出現(xiàn)顯著變化。120d批次的3.36mg Cd/kg組個(gè)別胎鼠出現(xiàn)趾骨骨化不全,頸椎椎體骨化不全或未骨化。所有胎鼠均未發(fā)現(xiàn)內(nèi)臟畸形。 5.仔鼠檢測(cè)結(jié)果：120d批次的3.36mg Cd/kg組的仔鼠體重與對(duì)照組相比明顯降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。其他批次染鎘組仔鼠體重與正常對(duì)照組相比未出現(xiàn)明顯變化,各批次中所有組別的雌性仔鼠占仔鼠總數(shù)的比例均無(wú)明顯差異(P0.05)。 6.窩平均活胎數(shù)統(tǒng)計(jì)結(jié)果：各批次各組別孕鼠的窩平均活胎鼠之間的差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論未成年雄性小鼠在連續(xù)多次短時(shí)間染鎘后,其成年后體內(nèi)雄激素水平下降,精子畸形率升高,睪丸鎘蓄積明顯并隨時(shí)間延長(zhǎng)而緩慢下降,附睪和睪丸病理學(xué)改變?cè)黾?此外,子代的體型和骨骼可出現(xiàn)發(fā)育延遲。
[Abstract]:Objective to explore the effects of cadmium exposure on reproductive system and fertility of immature male mice, and to accumulate basic data for further study on reproductive toxicity of cadmium.
Methods 96 4-week age SPF male NIH mice were randomly divided into normal control group, low, medium and high dose group, 24 rats in each group. A mixed solution of saline or cadmium chloride (CdCl2) and beta mercapto ethanol (ME) was injected into the abdominal cavity by the total dose of 0,0.84,1.68,3.36mg Cd/kg, respectively (the concentration ratio of two persons was 1:20), 1 times per day for 5 days. On the day of 30,60,90120, 6 animals were randomly selected from each group.
1. mating ability test: after the female and male mice were caged by 1:1 cage and the female rats were put in, the male mice were recorded for the first time to capture the female mice, that is to capture the incubation period and the number of female rats in the male rats within 20min.
2. male rats' reproductive toxicity test: (1) blood collection, centrifugation serum, serum testosterone (T). 2. Dissection brain, heart, lung, liver, spleen, kidney, testis, measurement of organ coefficient. 3. Leave right kidney, right testicle and part of liver, determine the content of cadmium in right testicle and part of the liver, and determine the content of cadmium in right epididymis (AAS). The left testis and the left epididymis were examined for pathology.
3. female mice mating rate and pregnancy rate test: the male and female rats were caged according to 1:2 cage, the day of vaginal suppository was recorded as zeroth days of pregnancy (GDO), and the mating rate of female rats was calculated after caged 12D. At GD14, the number of successful pregnant female rats was determined, and the rate of pregnancy was calculated.
4. fetal mice were detected in each group, and half of the pregnant female rats were taken out at GD17. General observation was carried out and the body length, tail length and body weight were measured. Then the skeletal deformity and visceral malformation were examined in the fetal rats.
5. rat detection: let the other half of the pregnant women give birth naturally. The offspring were weighed at 21 days old, and the sex was observed.
Statistics of the average number of live births in 6. litters: the average number of live births (including fetal rats and offspring rats) in each group of pregnant rats was statistically analyzed.
Results 1. the results of mating ability test showed that there was no significant change in mating ability between 30,60,90120d batches of cadmium exposed male rats and the control group.
2. male mice reproductive toxicity test results: (1) serum T detection results: the serum T content of the male rats in group 1.68,3.36mg Cd/kg of 30d, 60d and 120d batch was significantly lower than that of normal control (P0.05), and the serum T content of all the male mice in the 90d batch was significantly lower than that of the normal control group (P0.05). The dirty organ coefficient was significantly higher than that of the normal control group (P0.05). (3) the cadmium content of the organs of the male mice was significantly higher than that of the normal control group. The difference was statistically significant (P0.05). (4) the sperm abnormality rate: the sperm abnormality rate of the male mice with cadmium dyeing and the normal control group The difference was statistically significant (P0.05). (5) pathological examination of viscera: the pathological changes of kidney infected animals were mainly interstitial inflammatory cells infiltration, renal tubular epithelial cell degeneration, chronic pyelonephritis mainly. The pathological changes of liver were focal inflammatory cells infiltration, extramedullary hematopoiesis, liver cell nuclear swelling and so on. The pathological changes of the testis are mainly atrophied by some seminiferous tubules and some of the animals' testicular tissue is necrotic. The main pathological changes of epididymis are the disappearance of the sperm in the epididymal tube and the exfoliated spermatogenic cells in some epididymal tubes.
3. the test results of mating rate and conception rate of female rats: there was no significant change in mating rate and conception rate of female rats in each batch of cadmium treated group.
The test results of 4. fetal mice: the body length, tail length of each batch of cadmium stained fetal mice and the body weight were not significantly different from that of the normal control group. The 3.36mg Cd/kg group of the group of.120d had no bone ossification, the vertebral body ossification or no ossification. All the fetal rats had no visceral malformation.
5. mouse test results: the weight of the 3.36mg Cd/kg group of the 120d batch was significantly lower than the control group, the difference was statistically significant (P0.05). The weight of the other batch CD group had no significant change compared with the normal control group, and there was no significant difference in the proportion of the total number of the female offspring of all the groups in each batch (P0.05).
Statistical results of the average number of live births in 6. litters: there was no significant difference in the average litter size among pregnant rats in each batch (P0.05).
Conclusion the level of androgen in adult male mice decreased, the rate of sperm abnormality increased, and the accumulation of cadmium in testicles decreased slowly with time, and the pathological changes of epididymis and testis increased in adult male mice after several short periods of time.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R114
本文編號(hào)：2135739
[Abstract]:Objective to explore the effects of cadmium exposure on reproductive system and fertility of immature male mice, and to accumulate basic data for further study on reproductive toxicity of cadmium.
Methods 96 4-week age SPF male NIH mice were randomly divided into normal control group, low, medium and high dose group, 24 rats in each group. A mixed solution of saline or cadmium chloride (CdCl2) and beta mercapto ethanol (ME) was injected into the abdominal cavity by the total dose of 0,0.84,1.68,3.36mg Cd/kg, respectively (the concentration ratio of two persons was 1:20), 1 times per day for 5 days. On the day of 30,60,90120, 6 animals were randomly selected from each group.
1. mating ability test: after the female and male mice were caged by 1:1 cage and the female rats were put in, the male mice were recorded for the first time to capture the female mice, that is to capture the incubation period and the number of female rats in the male rats within 20min.
2. male rats' reproductive toxicity test: (1) blood collection, centrifugation serum, serum testosterone (T). 2. Dissection brain, heart, lung, liver, spleen, kidney, testis, measurement of organ coefficient. 3. Leave right kidney, right testicle and part of liver, determine the content of cadmium in right testicle and part of the liver, and determine the content of cadmium in right epididymis (AAS). The left testis and the left epididymis were examined for pathology.
3. female mice mating rate and pregnancy rate test: the male and female rats were caged according to 1:2 cage, the day of vaginal suppository was recorded as zeroth days of pregnancy (GDO), and the mating rate of female rats was calculated after caged 12D. At GD14, the number of successful pregnant female rats was determined, and the rate of pregnancy was calculated.
4. fetal mice were detected in each group, and half of the pregnant female rats were taken out at GD17. General observation was carried out and the body length, tail length and body weight were measured. Then the skeletal deformity and visceral malformation were examined in the fetal rats.
5. rat detection: let the other half of the pregnant women give birth naturally. The offspring were weighed at 21 days old, and the sex was observed.
Statistics of the average number of live births in 6. litters: the average number of live births (including fetal rats and offspring rats) in each group of pregnant rats was statistically analyzed.
Results 1. the results of mating ability test showed that there was no significant change in mating ability between 30,60,90120d batches of cadmium exposed male rats and the control group.
2. male mice reproductive toxicity test results: (1) serum T detection results: the serum T content of the male rats in group 1.68,3.36mg Cd/kg of 30d, 60d and 120d batch was significantly lower than that of normal control (P0.05), and the serum T content of all the male mice in the 90d batch was significantly lower than that of the normal control group (P0.05). The dirty organ coefficient was significantly higher than that of the normal control group (P0.05). (3) the cadmium content of the organs of the male mice was significantly higher than that of the normal control group. The difference was statistically significant (P0.05). (4) the sperm abnormality rate: the sperm abnormality rate of the male mice with cadmium dyeing and the normal control group The difference was statistically significant (P0.05). (5) pathological examination of viscera: the pathological changes of kidney infected animals were mainly interstitial inflammatory cells infiltration, renal tubular epithelial cell degeneration, chronic pyelonephritis mainly. The pathological changes of liver were focal inflammatory cells infiltration, extramedullary hematopoiesis, liver cell nuclear swelling and so on. The pathological changes of the testis are mainly atrophied by some seminiferous tubules and some of the animals' testicular tissue is necrotic. The main pathological changes of epididymis are the disappearance of the sperm in the epididymal tube and the exfoliated spermatogenic cells in some epididymal tubes.
3. the test results of mating rate and conception rate of female rats: there was no significant change in mating rate and conception rate of female rats in each batch of cadmium treated group.
The test results of 4. fetal mice: the body length, tail length of each batch of cadmium stained fetal mice and the body weight were not significantly different from that of the normal control group. The 3.36mg Cd/kg group of the group of.120d had no bone ossification, the vertebral body ossification or no ossification. All the fetal rats had no visceral malformation.
5. mouse test results: the weight of the 3.36mg Cd/kg group of the 120d batch was significantly lower than the control group, the difference was statistically significant (P0.05). The weight of the other batch CD group had no significant change compared with the normal control group, and there was no significant difference in the proportion of the total number of the female offspring of all the groups in each batch (P0.05).
Statistical results of the average number of live births in 6. litters: there was no significant difference in the average litter size among pregnant rats in each batch (P0.05).
Conclusion the level of androgen in adult male mice decreased, the rate of sperm abnormality increased, and the accumulation of cadmium in testicles decreased slowly with time, and the pathological changes of epididymis and testis increased in adult male mice after several short periods of time.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R114
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