异补骨脂素对维生素A缺乏大鼠胚胎上颈椎发育异常的影响  

Influence of Isopsoralen Supplement on Upper Cervical Malformation of Vitamin A Deficiency Rats Embryo

作　　者：麻昊宁[1,2] 刘楚吟[2,3] 朱宝[2,3] 董春科 姜良海[1,2] 谭明生[1,2] 

机构地区：[1]北京协和医学院研究生院,北京100730 [2]中日友好医院 [3]北京中医药大学研究生院

出　　处：《中国中医骨伤科杂志》2016年第1期4-8,共5页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：国家自然科学基金(81173423)

摘　　要：目的:研究雌鼠孕期维生素A(Vitamin A,VA)缺乏补充异补骨脂素对胎鼠上颈椎发育异常的影响及相关基因的表达。方法:初断乳(3周龄)SD雌性大鼠50只,按体质量顺序编号并随机分为维生素A缺乏(Vitamin A Deficiency,VAD)组及正常(Normal,N)组,分别喂养维生素A缺乏饲料和正常纯化饲料。喂养10周验证VAD组VA缺乏状态后,所有雌鼠按2∶1与正常雄鼠合笼,VAD组进一步分为维生素A缺乏空白组(VAD)、异补骨脂素补充低剂量组(Isopsoralen Supplement with Low Dose,ISL)、中剂量组(Isopsoralen Supplement with Medium Dose,ISM)、高剂量组(Isopsoralen Supplement with High Dose,ISH),其中ISL,ISM,ISH三组分别于妊娠0.5d起补充异补骨脂素。于妊娠第11.5天随机处死半数孕鼠取其胚胎,采用RT-QPCR方法分别检测胚胎Hoxd3基因和视黄酸受体(Retinoic Acid Receptors,RARs)蛋白mRNA分子表达水平。妊娠第21天取新生鼠,骨骼双染法观察上颈椎骨骼发育形态。结果:VAD组及异补骨脂素补充组均可见胚胎骨骼发育迟缓,上颈椎出现骨骼畸形早期表现。VAD组及异补骨脂素补充组Hoxd3基因及RARs mRNA表达水平下降(P<0.01),异补骨脂素补充组Hoxd3基因mRNA表达水平较VAD组高(P<0.05)且与补充剂量成正比,但差异不明显(P>0.05),RARs mRNA的表达与异补骨脂素的补充相关性不明显。结论:孕期雌鼠维生素A水平的下降可以导致胎鼠上颈椎一定程度的发育异常,而孕期补充异补骨脂素可上调Hoxd3基因的表达水平,但对形态异常的改善影响较小。Objective： To investigate the influence of isopsoralen supplement during gestation on upper cervical malformation in Vitamin A deficiency （VAD） rats embryo, and to explore the expression of relative genes. Methods： All 50 weaning female Spraguc Dawley rats were randomized divided into Vitamin A deficiency group and normal group according to body weights. The two groups were fed with normal purified diet and Vitamin A deficiency diet. After 10 weeks and confirming VAD status, all the fe male rats were mated with male rats （2： 1）. The VAD group was divided into VAD group, isopsoralen supplement with low dose group （ISL）, isopsoralen supplement with medium dose group （1SM）, isopsoralen supplement with high dose group （1SH）. lsopsoralen was supplied to ISL, ISM and ISH group since 0.5th gestational day. Half of the pregnant rats were sacrificed at 11. 5th gestational day and the embryos were taken to detect the expression of Hoxd3 gene and relinoic acid receptors （RARs） mRNA by using RT-QPCR. Neonatal rats were taken at 21th gestational day to observe morphology of craniovertebral junction using compound staining method. Results： Developmental retardation of bones and initial malformation of craniovertebral junction could be seen in VAD, ISL, ISM and ISH groups. The expression of Hoxd3 gene and RARs mRNA was decreased （P〈0.01）.The expression of Hoxd3 gene mRNA were higher in isopsoralen supplement groups compared with VAD group significantly （P d0.05）, and were directly proportional to the dose of isop.soralen, but there was no significance in different dose groups （P〉 0.05）. The expressions of RARs mRNA had no relations with isopsoralen supplement. Conclusion： Vilamin A deficiency during gestation causes malformation of eraniovertebral junction in rats＇ embryo. Supplement of isopsoralen during gestation improve the expression of Hoxd3 gene, but has little effect on morphology.

关 键 词：维生素A 异补骨脂素 上颈椎 Hoxd3基因 

分 类 号：R-33[医药卫生]