摘要
应用大鼠全胚胎培养(wholeembryoculture,WEC)方法,观察了黄花棘豆(Oxytropisochrocephala)主要有毒成分生物碱对体外胚胎生长发育、形态分化的影响。首先用醇类溶剂提取法从黄花棘豆中提取生物碱,薄板层析鉴定至少有4种生物碱成分。然后用体内同期胚胎对照及丝裂霉素C(MMC)阳性对照确证了所建立的WEC方法的可行性。在选定0.1%DMSO为该生物碱溶剂的基础上,分别在含有25、50、100、200mg/L生物碱的大鼠血清中培养9.5d胚胎48h,结果,在50mg/L以上可影响胚胎生长发育和器官分化,且呈剂量反应关系,主要表现为头长、颅臀长、体节数、蛋白质含量及17项形态学总记分低于溶剂对照组,而且随剂量增加,胚胎畸形也增多,200mg/L时畸形率为80%,主要表现为小头、后脑水肿、无听泡、无视泡、心包积液、肢芽缺失、尾异常、翻转异常等。生物碱还可影响脏壁卵黄囊(VYS)的血液循环,抑制其生长。结果提示,黄花棘豆生物碱对体外培养大鼠胚胎有直接发育毒性,VYS功能障碍也是引起胚胎发育毒性的重要机制。
The effects of Oxytropis ochrocephala alkaloid (total compositions) on growth development and morphological differentiation of rat embryo in vitro were studied by using the whole embryo culture (WEC) method. On the basis of determining 0.1% DMSO as solvent, embryos of day 9 5 were cultured in rat serum with the alkaloid of 25, 50, 100 and 200 mg/L, respectively. The result showed that the alkaloid could induce remarkable retardation of growth development and morphogensis at dose of over 50 mg/L. Some parameters, including head length, crown rump length,somite, protein content and score of morphology ,were lower than those of solvent control group. Dose dependent relationship were noticed.At concentration 200 mg/L, the rate of malformation embryo was 80%. The mainly abnormalities were microcephaly , swollen hind brain, retarded development of otic and optic vesicle, pericardial edema, limb bud hypoplasia, abnormal axial rotation, tail abnormal. Besides, the alkaloid could effect on blood circulation and inhibit growth of the visceral yolk sac(VYS) . The results indicated that O.ochrocephala alkaloid induced direct developmental toxicity on rat embryo in vitro. The barrier to function of the VYS may also be an important mechanism of the alkaloid on the embryo developmental toxicity.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1999年第1期55-60,共6页
Chinese Journal of Veterinary Science
关键词
黄花棘豆
生物碱
胚胎发育毒性
大鼠
xytropis ochrocephala
alkaloid
whole embryo culture
developmental toxicity
rat