肛门直肠畸形患儿血清维生素A水平及维甲酸受体α表达模式异常。
Abnormal serum vitamin A levels and retinoic acid receptor α expression patterns in children with anorectal malformation.
作者信息
Wang Zhili, Wang Quan, Gu Chengchao, Zhang Jingjie, Wang Yi
机构信息
Department of Neonatal Gastrointestinal Surgery, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, No. 136, Zhongshan 2nd Road, Yuzhong District, Chongqing, 400014, China.
Department of Cardiothoracic Surgery, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, No. 136, Zhongshan 2nd Road, Yuzhong District, Chongqing, 400014, China.
出版信息
Pediatr Surg Int. 2019 Aug;35(8):903-910. doi: 10.1007/s00383-019-04495-0. Epub 2019 Jun 12.
BACKGROUND
Anorectal malformation (ARM) is known to be associated with maldevelopment of the enteric nervous system (ENS), and vitamin A (VA) and its metabolite retinoic acid (RA) play important roles in ENS development. Thus, our aim was to investigate serum VA levels in ARM newborns and RA receptor (RAR) expression in the rectum of ARM patients and animal models.
METHODS
Serum VA concentrations were detected in newly diagnosed ARM neonates (n = 32) and neonates with non-alimentary tract malformations (n = 30). Intestinal specimens were divided into three groups: rectum from ARM patients (n = 30), colon from a stoma (n = 30) and rectum from controls (n = 4). RAR mRNA expression was evaluated by RT-qPCR. Rectum specimens from ARM patients were divided into two groups by postoperative pathology: the normal and lesion ganglion cell groups. Immunohistochemistry and Western blot were employed to detect RARα protein expression in rectum specimens. In addition, the ARM mouse model was induced by all-trans retinoid acid (ATRA), and the expression levels of RARα and the neuronal marker NeuN in the rectum of mice on embryonic day 16.5-18.5 (E16.5-18.5) were investigated.
RESULTS
The serum concentration of VA in ARM neonates was lower than that in control neonates (P < 0.0001), and RARα mRNA expression was lower in the rectum specimens from ARM patients than in the colon specimens from a stoma and the rectum specimens from controls (P < 0.05); there was no significant difference between the colon from a stoma and the rectum from controls. RARα protein was expressed in the nucleus of ganglion cells and nerve fibers, and RARα protein expression in the lesion ganglion cell group was significantly lower than that in the normal ganglion cell group (P < 0.01). Compared with the control mice, ARM mice at E16.5-18.5 showed decreased fluorescence intensity of RARα and NeuN in the rectum. RARα and NeuN mRNA expression in the rectum on E16.5-18.5 was lower in ARM mice than in control mice (P < 0.05).
CONCLUSION
Serum VA concentration and the RARα expression pattern are abnormal in the rectum in ARM and may contribute to the ENS maldevelopment in ARM.
背景
已知肛门直肠畸形（ARM）与肠神经系统（ENS）发育异常有关，维生素A（VA）及其代谢产物视黄酸（RA）在ENS发育中起重要作用。因此，我们的目的是研究ARM新生儿的血清VA水平以及ARM患者和动物模型直肠中RA受体（RAR）的表达。
方法
检测新诊断的ARM新生儿（n = 32）和非消化道畸形新生儿（n = 30）的血清VA浓度。肠道标本分为三组：ARM患者的直肠（n = 30）、造口结肠（n = 30）和对照组直肠（n = 4）。通过RT-qPCR评估RAR mRNA表达。根据术后病理将ARM患者的直肠标本分为两组：正常神经节细胞组和病变神经节细胞组。采用免疫组织化学和蛋白质印迹法检测直肠标本中RARα蛋白表达。此外，用全反式维甲酸（ATRA）诱导建立ARM小鼠模型，研究胚胎第16.5至18.5天（E16.5 - 18.5）小鼠直肠中RARα和神经元标志物NeuN的表达水平。
结果
ARM新生儿的血清VA浓度低于对照新生儿（P < 0.0001），ARM患者直肠标本中RARα mRNA表达低于造口结肠标本和对照直肠标本（P < 0.05）；造口结肠与对照直肠之间无显著差异。RARα蛋白在神经节细胞核和神经纤维中表达，病变神经节细胞组中RARα蛋白表达明显低于正常神经节细胞组（P < 0.01）。与对照小鼠相比，E16.5 - 18.5的ARM小鼠直肠中RARα和NeuN的荧光强度降低。ARM小鼠E16.5 - 18.5直肠中RARα和NeuN mRNA表达低于对照小鼠（P < 0.05）。
结论
ARM患者直肠中血清VA浓度和RARα表达模式异常，可能导致ARM中ENS发育异常。
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