[1]刘楚吟,麻昊宁,朱宝,等.维生素A缺乏导致新生大鼠枕颈部畸形的机制研究[J].中医正骨,2015,27(09):3-8.
　LIU Chuyin,MA Haoning,ZHU Bao,et al.Research on the mechanism of occipito-cervical deformity induced by vitamin A deficiency in neonatal rats[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(09):3-8.
点击复制

维生素A缺乏导致新生大鼠枕颈部畸形的机制研究()
分享到：

《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第27卷
期数:
2015年09期
页码:
3-8
栏目:
基础研究
出版日期:
2015-09-30

文章信息/Info

Title:
Research on the mechanism of occipito-cervical deformity induced by vitamin A deficiency in neonatal rats
作者:
刘楚吟1; 麻昊宁2; 朱宝1; 董春科1; 颜勤华1; 谭明生3
1.北京中医药大学,北京 100029;
2.北京协和医学院,北京 100730;
3.中日友好医院,北京 100029
Author(s):
LIU Chuyin1; MA Haoning2; ZHU Bao1; DONG Chunke1; YAN Qinhua1; TAN Mingsheng3
1.Beijing University of Chinese Medicine,Beijing 100029,China
2.Peking Union Medical College,Beijing 100730,China
3.China-Japan Friendship Hospital,Beijing 100029,China
关键词:
维生素A缺乏;  枕骨;  颈椎;  先天畸形;  Hoxd3基因;  动物实验
Keywords:
vitamin A deficiency;  occipital bone;  cervical vertebrae;  congenital abnormalities;  Hoxd3 gene;  animal experimentation
摘要:
目的:探讨维生素A缺乏导致新生大鼠枕颈部畸形的作用机制。方法:将40只5周龄雌性健康SPF级SD大鼠随机分为2组,正常对照组20只,维生素A缺乏组20只; 20只适龄雄性健康SPF级SD大鼠分笼饲养。正常对照组饲以AIN-93M饲料,维生素A缺乏组饲以缺乏维生素A的AIN-93M饲料,雄性大鼠饲以AIN-93M饲料。饲养至第10周从雌性大鼠的目外眦取血,采用酶联免疫吸附法检测2组雌鼠血清维生素A含量,鉴定大鼠模型。造模成功后,将大鼠按雌2雄1的比例合笼交配,第2天清晨检查阴栓,有阴栓者记为E0 d。于妊娠E11.5 d取2组半数的孕鼠,麻醉处死后剖腹,每只孕鼠取出2只胚胎,剥离被膜,置于液氮中,然后转移至-80 ℃的冰箱里保存待测。于妊娠E21 d取新生鼠,麻醉处死后剥离皮肤筋膜以及内脏,置于95%的酒精中固定待测。采用RT-qPCR方法检测胚胎Hoxd3基因mRNA表达情况; 采用双色染色法制作大鼠骨骼标本,观察新生鼠枕颈部骨骼发育情况。结果:①雌鼠血清维生素A含量。维生素A缺乏组大鼠血清维生素A含量明显低于正常对照组[(214±39)ng·mL-1,(432±54)ng·mL-1,t=14.636,P=0.000]。②胚胎Hoxd3基因mRNA的表达量。维生素A缺乏组胚胎Hoxd3基因表达量低于正常对照组(0.58±0.11,1.03±0.14,t=10.407,P=0.000)。③新生鼠枕颈部骨骼形态。正常对照组可见枕骨与寰枢椎发育正常,对位关系良好; 维生素A缺乏组可见寰椎后弓与枕骨大孔后缘角度增大,寰枢椎对位关系异常,寰枢椎后弓角度增大,C2～C7椎弓及棘突高度降低、宽度变窄,且相邻椎体间隙增大。结论:维生素A缺乏可以导致新生大鼠枕颈部结构发育异常,这可能与其胚胎发育期Hoxd3基因mRNA表达水平下降有关,但其具体的分子生物学机制还有待进一步研究。
Abstract:
Objective:To explore the mechanism of occipito-cervical deformity induced by vitamin A deficiency in neonatal rats.Methods:Forty healthy 5-week-old female SPF-grade SD rats were randomly divided into 2 groups,20 rats in each group.The female rats were respectively fed with normal AIN-93M stoyer(normal control group)and AIN-93M stoyer lacking in vitamin A(vitamin A deficiency group,VAD group).Meanwhile,20 healthy male SPF-grade SD rats were recruited and fed with AIN-93M stoyer in separate cages.Ten weeks later,serum content of vitamin A were detected in female rats by enzyme-linked immunosorbent assay to identify the rat models.After successful modeling,the rats were mated according to the female-male ratio of 2:1.In the next morning,the female rats were examined in vaginal plug and rats which had vaginal plug were recorded as E0 d.At 11.5 days after gestation,half of the pregnant rats were anesthetized and sacrificed.The belly of each pregnant rat were cut open and two embryos were fetched out and their capsules were stripped.The embryos were placed in liquid nitrogen and then were transferred to refrigerator and reserved at -80 ℃.On 21st gestational day,neonatal rats were anesthetized and sacrificed.Their skins and fascias were stripped and their internal organs were removed.Then the neonatal rats were reserved in 95% alcohol.The qRT-PCR assays was used for detecting the Hoxd3 mRNA expression in embryos and the bicolor staining method was used for observing the development of occipito-cervical bone of neonatal rats.Results:The serum contents of vitamin A were obviously lower in the VAD group compared to the control group(214+/-39 vs 432+/-54 ng/mL,t=14.636,P=0.000).The Hoxd3 mRNA expression in embryos was obviously lower in the VAD group compared to the control group(0.58+/-0.11 vs-- 1.03+/-0.14,t=10.407,P=0.000).Normally developed occipital bone,atlas and axis and right contraposition of atlanto-axial joint were found in the control group.However,in the VAD group the angle betwwen posterior arch of atlas and posterior margin of great occipital foramen increased,and contraposition of atlanto-axial joint was abnormal.Atlantoaxial angle and intervertebral space increased,while the height and width of vertebral arch and spinous process of C2-C7 decreased.Conclusion:Vitamin A deficiency can lead to occipito-cervical dysplasia in neonatal rats,which may be related to decrease in Hoxd3 mRNA expression in embryonic development period.However,the specific molecular mechanism needs to be further studied.

参考文献/References:

[1] Clagett-Dame M,Knutson D.Vitamin a in reproduction and development[J].Nutrients,2011,3(4):385-428.
[2] Li N,Sun S,Wang D,et al.Suppression of retinoic acid receptors May contribute to embryonic skeleton hypoplasia in maternal rats with chronic vitamin A deficiency[J].J Nutr Biochem,2010,21(8):710-716.
[3] 郑怀竞,韩松.临床检验ELISA指南[M].北京:北京医科大学中国协和医科大学联合出版社,1994:39-52.
[4] 赵荧,张栩胤.整体显示大鼠骨骼和软骨的复合染色法[J].解剖学报,2006,37(1):117-119.
[5] Niwa Y,Shimojo H,Isomura A,et al.Different types of oscillations in Notch and Fgf signaling regulate the spatiotemporal periodicity of somitogenesis[J].Genes Dev,2011,25(11):1115-1120.
[6] Pourquié O.Segmentation of the paraxial mesoderm and vertebrate somitogenesis[J].Curr Top Dev Biol,2000,47(5):81-105.
[7] 贾连顺,李家顺.枕颈部外科学[M].上海:上海科学技术出版社,2003:3.
[8] 谭明生,张光铂.上颈椎外科学[M].北京:人民卫生出版社,2010:334.
[9] Iimura T,Denans N,Pourquié O.Establishment of Hox vertebral identities in the embryonic spine precursors[J].Curr Top Dev Biol,2009,88:201-234.
[10] Juan AH,Ruddle FH.Enhancer timing of Hox gene expression: deletion of the endogenous Hoxc8 early enhancer[J].Development,2003,130(20):4823-4834.
[11] Wellik DM,Capecchi MR.Hox10 and Hox11 genes are required to globally pattern the mammalian skeleton[J].Science,2003,301(5631):363-367.
[12] Carapuço M,Nóvoa A,Bobola N,et al.Hox genes specify vertebral types in the presomitic mesoderm[J].Genes Dev,2005,19(18):2116-2121.
[13] Pang D,Thompson DN.Embryology and bony malformations of the craniovertebral junction[J].Childs Nerv Syst,2011,27(4):523-564.
[14] Condie BG,Capecchi MR.Rats homozygous for a targeted disruption of Hoxd-3(Hox-4.1)exhibit anterior transformations of the first and second cervical vertebrae, the Atlas and the axis[J].Development,1993,119(3):579-595.
[15] Condie BG,Capecchi MR.Rats with targeted disruptions in the paralogous genes hoxa-3 and hoxd-3 reveal synergistic interactions[J].Nature,1994,370(6487):304-307.
[16] Manley NR,Capecchi MR.Hox group 3 paralogous genes act synergistically in the formation of somitic and neural crest-derived structures[J].Dev Biol,1997,192(2):274-288.
[17] 金淑清,浦予飞,裘莹.HOX基因的研究进展[J].癌症进展,2011,9(2):154-158.
[18] Kaiser ME,Merrill RA,Stein AC,et al.Vitamin a deficiency in the late gastrula stage rat embryo results in a one to two vertebral anteriorization that extends throughout the axial skeleton[J].Dev Biol,2003,257(1):14-29.
[19] See AW,Kaiser ME,White JC,et al.A nutritional model of late embryonic vitamin A deficiency produces defects in organogenesis at a high penetrance and reveals new roles for the vitamin in skeletal development[J].Dev Biol,2008,316(2):171-190.
[20] 刘楚吟,谭明生,陈鹏明,等.维生素A缺乏对小鼠胚胎Hoxd3基因表达的影响[J].中国中医骨伤科杂志,2014,22(7):1-3.
[21] Kessel M,Gruss P.Homeotic transformations of murine vertebrae and concomitant alteration of Hox codes induced by retinoic acid[J].Cell,1991,67(1):89-104.
[22] Lohnes D,Mark M,Mendelsohn C,et al.Function of the retinoic acid receptors(RARs)during development(I).Craniofacial and skeletal abnormalities in RAR double mutants[J].Development,1994,120(10):2723-2748.
[23] Aulehla A,Pourquié O.Signaling gradients during paraxial mesoderm development[J].Cold Spring Harb Perspect Biol,2010,2(2):869.

相似文献/References:

[1]周树一,许楠健,王扬,等.后路经寰枕关节-枕骨髁-斜坡置钉技术的可行性和安全性研究[J].中医正骨,2022,34(12):8.
　ZHOU Shuyi,XU Nanjian,WANG Yang,et al.Feasibility and safety of posterior atlantooccipital joint-occipital condyle-clivus screw fixation[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2022,34(09):8.

备注/Memo

备注/Memo:
2015-06-08收稿 2015-07-08修回
基金项目:国家自然科学基金项目(81173423)
通讯作者:谭明生 E-mail:zrtanms@sina.com
更新日期/Last Update: 2015-09-30