【作者】 郭红刚；

【导师】 杨建一；

【作者基本信息】 山西医科大学 ， 遗传学， 2006， 硕士

【摘要】 丙烯酰胺(Acrylamide,AA)主要用于生产其聚合物一聚丙烯酰胺。聚丙烯酰胺是一种工业絮凝剂,广泛应用于饮水净化、污水处理、食品加工、造纸和采矿业。丙烯酰胺单体则在建造堤坝、地基、隧道和公路中用于生产水泥和土壤的稳定剂,它在粘合剂和纺织品制造业以及胶片冲印中也有应用。世界上,在工业加工中接触AA的工人数以万计,在分子生物学实验室,这一数值估计更多。2002年4月瑞典国家食品管理局和瑞典斯德哥尔摩大学同时报道:在一些常见煎烤烹制的食品中检测到较高含量的丙烯酰胺,这一发现引起了国际社会的普遍关注。丙烯酰胺在1994年已经被国际癌症机构(IARC)列为“人类可能致癌物”。丙烯酰胺不仅对动物有神经毒性,还有生殖毒性、胚胎毒性和致癌性。迄今为止,国内外研究的重点多集中在丙烯酰胺对动物和人类的致癌问题上。近年来,随着研究的深入,丙烯酰胺的生殖毒性逐渐受到关注。目前丙烯酰胺对啮齿类动物生殖毒性的原因不是十分清楚,可能的原因有①丙烯酰胺的代谢产物环氧丙烯酰胺连接到精细胞的鱼精蛋白上,影响了精子的形态,使显性致死实验阳性率升高。②丙烯酰胺连接到动力蛋白上,影响了神经远端轴突的转运,干扰了神经胞体和轴突之间的分子转运,导致下肢无力(可发展为轻瘫),可引起外生殖器感觉异常,影响了交配行为;丙烯酰胺也可能干扰了精子的鞭毛蛋白,影响了精子的运动。③丙烯酰胺的代谢产物环氧丙烯酰胺与DNA形成加合物,引起遗传物质损伤。为进一步阐明丙烯酰胺生殖毒性的原因,本研究以雄性小鼠的生殖细胞为研究对象,经腹腔注射丙烯酰胺建立动物模型,设阴性、阳性对照组和浓度为25mg/kg.bw、50mg/kg.bw、100mg/kg.bw丙烯酰胺剂量组,用单细胞凝胶电泳技术、组织病理学技术、生殖细胞早期精细胞微核技术、初级精母细胞染色体技术、精子畸形实验及放射免疫法联合评价丙烯酰胺对雄性小鼠的生殖毒性及遗传毒性,为进一步探讨其生殖损伤原因提供实验室数据。本研究结果提示丙烯酰胺染毒后睾丸细胞DNA损伤程度较淋巴细胞DNA损伤程度大;病理切片提示高剂量染毒(100mg/kg.bw,连续染毒天5)后可引起生殖细胞数目减少;生殖细胞早期精细胞微核率、初级精母细胞染色体畸变率、精子畸形率随着染毒剂量的增加而增加,均有一定的剂量依赖关系;丙烯酰胺染毒后对血清中睾酮浓度基本没有影响。本研究认为丙烯酰胺对睾丸细胞造成的遗传损伤较淋巴细胞大,提示睾丸细胞可能是丙烯酰胺的靶细胞之一;丙烯酰胺可减少曲细精管中精细胞的数量;引起生殖细胞染色体断裂和非整倍体数增加;引起精子畸形率增加。更多还原

【Abstract】 Acrylamide(AA)is used primarily for the production of polyacrylamide. Polyacrylamide have its largest market as flocculants for the purification of water, waste water treatment, food processing, paper and mineral processing, while the monomer is used to produce grouts and soil stabilizer in the construction of dams, foundations, tunnels and roadways. It also has some minor uses in the manufacture of adhesives, the textile industry, and some photographic applications. The number of workers exposed to AA in industrial processes may be several thousand worldwide and the population potentially exposed to AA in molecular biological laboratories is estimated to be as high as much.April in 2002,the Swedish National Food Authority and the Uiversity of Stockholm jointly announced that they had found considerable levels of acrylamide in starchbased foods. This discovery attracted people’s attentions. Because Acrylamide has been classified as“probably carcinogenic to human”by the International Agency for Rearch on Cancer (IARC) in 1994. The sufficient evidences shows that AA is known to produce a wide range of toxic efects, including neurotoxicity, genotoxicity, reproductive toxicity and carcinogenicity.Up to now , increasing attention has been given to its cancer inductive properties in domestic and abroad ,more recently, reproductive toxicity of acrylamide is paid attention to.The mechanism of action acrylamide toxicity is not known clearly,it appears to be①Glycidamide binding to spermatid protamines effects on sperm morphology and results in dominant lethality .②Acrylamide binding to the motor proteins kinesin and dynein results in distal axonopathy and interferes with molecular transport between the neuron cell body and axon and induces hindlimb weakness(progressing to paresis),perhaps also produce anesthesia to the penis, resulting in effects on mounting and intromission and effects on sperm motility(from interference with flagellar function ).③Glycidamide binding directly to DNA(widespread DNA adducts have been reported )likely causes mutation .In order to explore the reproductive toxicity of acrylamide,male mices were taken as subjects .The animal model was made by intraperitioneal injection with acrylamide,and the mices were divided into 5 groups,such as normal control,positive control、25mg/kg.bw、50mg/kg.bw and 100mg/kg.bw dosage. Detection of acrylamide induced DNA double strand breakage in mice germ cell and lymphocyte were made. The micronucleus rate of early spermatid was detected by micronucleus technique. Chromosome of primariry spermatocyte was detected by chromosome technique. Sperm parameter was detected by sperm aberration test .The serum concentration of Testosterone(T) was detected by radiommunoassay. Histological of更多还原