镉暴露对未成年雄性小鼠的生殖毒性研究
发布时间：2018-07-21 13:53
【摘要】：目的探索镉染毒对未成年雄性小鼠生殖系统的损伤及生育能力的影响,为进一步研究镉的生殖毒性积累基础资料。 方法96只四周龄SPF级雄性NIH小鼠随机分为正常对照组,低、中、高剂量组,每组24只。按照总剂量分别为0、0.84、1.68、3.36mg Cd/kg体重腹腔注射生理盐水或氯化镉(CdCl2)和β-巯基乙醇(ME)的混合溶液(二者浓度比为1：20),1次/天,连续5天。给药结束后的第30,60,90,120天,各组分别随机取6只动物,进行如下处理： 1.交配能力检测：将雌雄小鼠按1：1合笼,雌鼠放入后,记录雄鼠首次捕捉雌鼠的时间即捕捉潜伏期及20min内雄鼠捕捉雌鼠的次数。 2.雄鼠生殖毒性检测：①采血,离心取血清,测定血清睾酮(T)。②解剖取脑、心、肺、肝、脾、肾、睾丸,测脏器系数。③留取右肾、右睾丸和部分肝,用原子吸收光谱仪(AAS)测定其中的镉含量。④取右侧附睾观察精子畸形率。⑤固定肝脏、脾脏、左肾、左睾丸和左附睾进行病理检查。 3.雌鼠交配率和受孕率检测：将雄鼠与雌鼠按1：2合笼,发现阴道栓当天记为妊娠第0天(GDO),合笼12d后计算雌鼠交配率,在GD14,确定成功怀孕雌鼠的数量,计算受孕率。 4.胎鼠检测：每组随机取一半受孕雌鼠,于GD17处死取出胎鼠。进行一般观察,并测量体长、尾长及体重。再对胎鼠分别进行骨骼畸形和内脏畸形检查。 5.仔鼠检测：让每组另一半受孕雌鼠自然分娩,仔鼠21日龄时称重,并观察性别。 6.窝平均活胎数统计：对各批次各组孕鼠的窝平均活胎数(包括胎鼠和仔鼠)进行统计。 结果1.交配能力检测结果：30,60,90,120d批次染镉雄鼠的交配能力与对照组相比未出现显著变化。 2.雄鼠生殖毒性检测结果：①血清T检测结果：30d,60d,120d批次的1.68、3.36mg Cd/kg组雄鼠的血清T含量显著低于正常对照(P0.05),90d批次的所有染镉雄鼠血清T含量均显著低于正常对照组(P0.05)。②120d批次的0.84mg Cd/kg组肝脏、肺脏,1.68mg Cd/kg组肝脏的脏器系数较正常对照组显著升高,差异有统计学意义(P0.05)。③脏器镉含量测定结果：各批次染镉雄鼠肝、肾、睾丸镉含量均显著高于正常对照组,差异有统计学意义(P0.05)。④精子畸形率：各批次染镉雄鼠精子畸形率与正常对照组相比,均显著升高,差异有统计学意义(P0.05)。⑤脏器病理检查结果：染镉动物肾脏病理变化主要以间质炎细胞浸润,肾小管上皮细胞变性,慢性肾盂肾炎为主。肝脏的病理改变主要有：局灶性炎细胞浸润,肝脏髓外造血,肝细胞核肿胀等。脾脏也出现了髓外造血。睾丸的病理改变主要以部分生精小管萎缩,部分动物睾丸组织坏死为主。附睾的病理改变主要有：附睾管内精子消失,部分附睾管内可见脱落生精细胞。 3.雌鼠交配率和受孕率检测结果：各批次染镉组雌鼠的交配率和受孕率与对照组相比均未出现显著变化。 4.胎鼠检测结果：各批次染镉组胎鼠的体长、尾长、体重与正常对照组相比未出现显著变化。120d批次的3.36mg Cd/kg组个别胎鼠出现趾骨骨化不全,颈椎椎体骨化不全或未骨化。所有胎鼠均未发现内脏畸形。 5.仔鼠检测结果：120d批次的3.36mg Cd/kg组的仔鼠体重与对照组相比明显降低,差异有统计学意义(P0.05)。其他批次染镉组仔鼠体重与正常对照组相比未出现明显变化,各批次中所有组别的雌性仔鼠占仔鼠总数的比例均无明显差异(P0.05)。 6.窝平均活胎数统计结果：各批次各组别孕鼠的窝平均活胎鼠之间的差异均无统计学意义(P0.05)。 结论未成年雄性小鼠在连续多次短时间染镉后,其成年后体内雄激素水平下降,精子畸形率升高,睾丸镉蓄积明显并随时间延长而缓慢下降,附睾和睾丸病理学改变增加,此外,子代的体型和骨骼可出现发育延迟。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114
本文编号：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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114
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