摘要
为了探究母代大鼠孕前染砷(As_2O_3)对胎鼠睾丸前精原细胞的毒性作用及生殖发育潜能,将24只雌性SD大鼠随机分为3组(n=8),通过灌胃方式建立As_2O_3染毒模型后,按"一代一窝"繁殖试验技术获得后代。大鼠妊娠第17.5天时取材,碱性磷酸酶染色法鉴定睾丸前精原细胞,透射电镜技术观察前精原细胞超微结构变化,qRT-PCR检测生殖细胞发育相关基因vasa mRNA表达,从而分析前精原细胞的凋亡情况及发育潜能。试验结果显示,与对照组比较,随着孕前染As_2O_3剂量的增加,胞核染色质进一步凝缩成致密团块;粗面内质网减少,结构损害逐渐加重;线粒体肿胀、破裂增多,线粒体颗粒分布更少,电子密度进一步降低,呈空泡状坏死。试验组睾丸组织中vasa mRNA表达量下降,与对照组比较差异显著(P<0.05),vasa mRNA表达量下降程度与孕前染As_2O_3剂量呈一定的负相关。试验表明,雌性SD大鼠孕前所染As_2O_3可通过胎盘屏障进入胎儿血液循环,并穿透前精原细胞的胞膜和线粒体膜,影响线粒体结构发生改变,诱导前精原细胞发生凋亡,vasa mRNA表达量下降,从而引起胚胎生殖毒性,抑制生殖细胞发育。
To explore the toxic effects of pregestational arsenic on developmental potential of fetal rat testicular pre-primary cells, the 24 female SD rats were randomly divided into three groups（n=8）.Arsenic exposure model was established by means of gastric lavage,and offspring were obtained through ＂One generation nest＂ breeding test technology. In 17.5 days fetal rats,testicular pre-primary cells were identified by alkaline phosphatase staining method. The ultrastructural changes of testicular pre-primary cells were observed by transmission electron microscope. The expression of vasa mRNA was detected by RT-PCR,which was used for analysis of apoptosis and developmental potential of testicular pre-primary cells. Compared with the control group, with the increase of the dose of As2O3,nuclear chromatin further condensed into dense clumps,rough endoplasmic reticulum decreased,and structure damage was gradually increased. Mitochondria were swollen,the fracture was increased,the distribution was less,the electron density was decreased and the shape was necrosis. Compared with the control group,the expression of vasa mRNA in testicular tissue was decreased（P〈0.05）,and the expression of vasa mRNA and the dose of As2O3 was negative correlation. The arsenic exposed to pregnant female SD rats could enter the fetal circulation through the placenta barrier,piercd cell membrane and mitochondrial membrane of testicular pre-primary cells,and mitochondrial structures were changed,flowed by induction of apoptosis of testicular pre-primary cells. Expression of mRNA vasa were decreased,causing the embryo reproductive toxicity and inhibited germ cell development.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2016年第7期928-933,共6页
Chinese Veterinary Science
基金
贵州省科学技术基金项目(黔科合J字LKZ[2012]36号)
贵州省教育厅自然科学研究项目(黔教合KY字[2014]263)
贵州省卫生厅项目(gzwkj2011-1-028)
关键词
细胞凋亡
前精原细胞
线粒体
砷
cell apoptosis
pre-primary cell
mitochondria
arsenic
