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[1]劉楚吟,麻昊寧,朱寶,等.維生素A缺乏導(dǎo)致新生大鼠枕頸部畸形的機制研究[J].中醫(yī)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.
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維生素A缺乏導(dǎo)致新生大鼠枕頸部畸形的機制研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第27卷
期數(shù):
2015年09期
頁碼:
3-8
欄目:
基礎(chǔ)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:
劉楚吟1; 麻昊寧2; 朱寶1; 董春科1; 顏勤華1; 譚明生3
1.北京中醫(yī)藥大學(xué),北京 100029;
2.北京協(xié)和醫(yī)學(xué)院,北京 100730;
3.中日友好醫(yī)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
關(guān)鍵詞:
維生素A缺乏;  枕骨;  頸椎;  先天畸形;  Hoxd3基因;  動物實驗
Keywords:
vitamin A deficiency;  occipital bone;  cervical vertebrae;  congenital abnormalities;  Hoxd3 gene;  animal experimentation
摘要:
目的:探討維生素A缺乏導(dǎo)致新生大鼠枕頸部畸形的作用機制。方法:將40只5周齡雌性健康SPF級SD大鼠隨機分為2組,正常對照組20只,維生素A缺乏組20只; 20只適齡雄性健康SPF級SD大鼠分籠飼養(yǎng)。正常對照組飼以AIN-93M飼料,維生素A缺乏組飼以缺乏維生素A的AIN-93M飼料,雄性大鼠飼以AIN-93M飼料。飼養(yǎng)至第10周從雌性大鼠的目外眥取血,采用酶聯(lián)免疫吸附法檢測2組雌鼠血清維生素A含量,鑒定大鼠模型。造模成功后,將大鼠按雌2雄1的比例合籠交配,第2天清晨檢查陰栓,有陰栓者記為E0 d。于妊娠E11.5 d取2組半數(shù)的孕鼠,麻醉處死后剖腹,每只孕鼠取出2只胚胎,剝離被膜,置于液氮中,然后轉(zhuǎn)移至-80 ℃的冰箱里保存待測。于妊娠E21 d取新生鼠,麻醉處死后剝離皮膚筋膜以及內(nèi)臟,置于95%的酒精中固定待測。采用RT-qPCR方法檢測胚胎Hoxd3基因mRNA表達(dá)情況; 采用雙色染色法制作大鼠骨骼標(biāo)本,觀察新生鼠枕頸部骨骼發(fā)育情況。結(jié)果:①雌鼠血清維生素A含量。維生素A缺乏組大鼠血清維生素A含量明顯低于正常對照組[(214±39)ng·mL-1,(432±54)ng·mL-1,t=14.636,P=0.000]。②胚胎Hoxd3基因mRNA的表達(dá)量。維生素A缺乏組胚胎Hoxd3基因表達(dá)量低于正常對照組(0.58±0.11,1.03±0.14,t=10.407,P=0.000)。③新生鼠枕頸部骨骼形態(tài)。正常對照組可見枕骨與寰樞椎發(fā)育正常,對位關(guān)系良好; 維生素A缺乏組可見寰椎后弓與枕骨大孔后緣角度增大,寰樞椎對位關(guān)系異常,寰樞椎后弓角度增大,C2～C7椎弓及棘突高度降低、寬度變窄,且相鄰椎體間隙增大。結(jié)論:維生素A缺乏可以導(dǎo)致新生大鼠枕頸部結(jié)構(gòu)發(fā)育異常,這可能與其胚胎發(fā)育期Hoxd3基因mRNA表達(dá)水平下降有關(guān),但其具體的分子生物學(xué)機制還有待進(jìn)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.

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備注/Memo

備注/Memo:
2015-06-08收稿 2015-07-08修回
基金項目:國家自然科學(xué)基金項目(81173423)
通訊作者:譚明生 E-mail:[email protected]
更新日期/Last Update: 2015-09-30