摘要
目的:探讨羟基脲(hydroxyurea,HU)诱导大鼠生精细胞凋亡发生的量效关系和时效关系。方法:量效关系研究中,雄性Wistar大鼠25只,分为5组,每组5只,4组为试验组,1组为对照组,4个试验组分别腹腔注射HU100mg/kg、200mg/kg、400mg/kg和600mg/kg,对照组给予磷酸盐缓冲液2ml/kg,给药后12h处死;时效关系研究中,雄性Wistar大鼠20只,分为4组,每组5只,3组均腹腔注射HU400mg/kg,1组为空白对照,分别于给药后6、12和24h处死大鼠。所有大鼠处死后称体重,取睾丸组织并称重,Bouin's液固定后,进行HE染色、原位细胞凋亡检测(TUNEL)、糖原PAS染色,光学显微镜下观察组织结构,计数凋亡细胞并进行生精周期阶段的分析。结果:给药后6、12、24h大鼠体重、睾丸重量与给药前相比,未见明显变化。量效关系研究中,大鼠生精细胞凋亡阳性小管所占比率和凋亡指数随着HU剂量的增加而明显增加,其中以400mg/kg为最高,分别为(38.7±2.0)%和(496.4±66.8)(P<0.01)。时效关系研究中,与对照组比较,给药后12h的每管凋亡阳性生精细胞数和凋亡指数达峰值,分别为(12.9±2.10)和(496.4±66.8)(P<0.01);TUNEL阳性细胞主要存在于生精周期的Ⅰ-Ⅳ时相。结论:HU诱导的大鼠睾丸生精细胞凋亡具有细胞特异性和阶段特异性。
Objective： To study the dose-effect and time-effect relations to spermatogenic cell apoptosis of rats induced by hydroxyurea. Methods：In the dose-effect study, 25 male Wistar rats were divided into 5 groups （5 rats in each groups）. Of the 5 groups, 4 were the test groups and 1 was the control group. Each of the four test groups was administered with a different dose of hydroxyurea （ 100 mg/kg, 200 mg/kg, 400 mg/kg, and 600 mg/kg） by intraperitoneal injection, respectively. The control group was administered with phosphate-buffered solution 2 ml/kg. All rats were killed 12 hours after dosing. In the time-effect study, 20 male Wistar rats were divided into 4 groups（5 rats in each groups）. Of the 4 groups, 3 were the test groups and I was the blank control group. The three test groups were administered with hydroxyurea 400 mg/kg by intraperitoneal injection. All rats were killed 6, 12, and 24 hours after dosing. The dead rats and their testes were weighed. The testes were fixed in Bouing solution and then stained with hematoxylin-eosin. Morphological changes were observed under light microscope. The TUNEL staining of apoptotic cells and PAS staining for glycogen were performed. The number of apoptotic cells was counted and the stages of seminiferous epithelial cycle were analysed. Results： There were no significant changes in weights of body and testis 6, 12, and ：24 hours after dosing compared with before dosing. The results of the dose-effect study showed the rates of apoptotic-positive seminiferous tubule and apoptotic index were markedly elevated with the increase in doses, especially in 400 mg/kg [ （38.7 ± 2.0） % and （496.4 ± 66.8 ）, respectively ] （ P 〈 0. 01 ）. The results of the time-effect study showed the mean number of apoptotic-positive spermatogenic cell and apoptotic index reached a peak at 12 hours after dosing [ （12.9 ± 2. 10） and （496.4±66.8）, respectively] compared with the control group. TUNEL-positive cells were mainly in stage Ⅰ - Ⅳ of seminiferous cycle. Conclusion： The spermatogenic cell apoptosis of rats induced by hydroxyurea is cell-specific and stage-specific.
出处
《药物不良反应杂志》
2008年第6期400-405,共6页
Adverse Drug Reactions Journal
基金
"国家高新技术研究发展计划"资助项目(863计划
2002AA2Z342D)
关键词
羟基脲
大鼠
生殖毒性
凋亡
生精上皮周期
hydroxyurea
rats
reproductive toxicity
apoptosis
seminiferous epithelial cycle
作者简介
通讯作者：廖明阳，男，研究员，博士生导师，Tel：（010）66931628，E—mail：liaomingy@hotmail．com
