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先天性肠无神经节细胞症患儿中血清维生素A及肠组织维甲酸受体α和HA117的表达特点
罗媛圆滕银平刘行李力金先庆*
解剖学报 ›› 2015, Vol. 46 ›› Issue (4) : 558-563.
先天性肠无神经节细胞症患儿中血清维生素A及肠组织维甲酸受体α和HA117的表达特点
Expression of vitamin A in peripheral blood and differential expression of retinoic acid receptor α and HA117 in colon segments of children with Hirschsprung’s disease
目的 检测先天性肠无神经节细胞症（HD）患儿中血清维生素A及肠组织维甲酸受体α（RARα）、抗分化长链非编码RNA（LncRNA）HA117的表达特点，探讨其与HD发病机制的可能联系。方法 收取17例临床诊断为HD的患儿术前外周血及手术结肠组织标本，结肠组织由近端至远端分为3组：吻合口组、扩张段组和痉挛段组。检测血清维生素A浓度，采用Real-time PCR、免疫组织化学染色、Western blotting方法检测RARα及HA117在各组结肠组织的表达水平。结果 17例HD患儿均有不同程度维生素A缺乏，血清维生素A平均浓度为（0.73±0.05）μmol/L。在吻合口段，RARα在肌间神经丛可见棕黄色的阳性染色，而在痉挛段几乎未见阳性神经丛染色；在吻合口组、扩张段组和痉挛段组，RARα mRNA相对表达量依次为3.62 ± 1.12、1.81 ± 0.69、0.65 ± 0.32，与吻合口相比，痉挛段表达量降低（P＜0.05, n=17）；HA117 mRNA相对表达量依次为0.47 ± 0.15、1.25 ± 0.40、2.57 ± 0.70，与吻合口相比，痉挛段表达量增高（P＜0.001, n=17）；RARα蛋白相对表达量依次为1.16 ± 0.05、0.99 ± 0.05、0.88 ± 0.04，与吻合口相比，扩张段及痉挛段表达量降低（分别为P＜0.05、P＜0.001,n=17）。 结论 维生素A、RARα及HA117可能共同参与肠神经系统（ENS）的发育过程；其中，维生素A和RARα可能促进了ENS的正常分化发育，而高表达的HA117可能对抗了前者的促分化作用，阻碍了ENS正常分化发育过程，进而导致HD的发生。
Objective To describe the expression profiles of vitamin A in blood serum and retinoic acid receptor α（RARα）, HA117 in different portions of the colon of patients diagnosed with Hirschsprung’s disease (HD). Methods Peripheral blood and colon specimens were collected from 17 HD patients. The concentration of vitamin A was detected in blood serum, and the colon specimens were grouped into 3 segments: anastomosis, dilated segment and stenotic segment. Levels of RARα mRNA and HA117 RNA were evaluated by Realtime PCR. RARα protein expression were analyzed by immunohistochemistry and Western blotting. Results The concentration of vitamin A in peripheral blood is (0.73±0.05) μmol/L, which was below the normal level. The levels of RARα mRNA expression was significantly lower in the stenotic segment compared to proximal anastomosis (P＜0.05,n=17). The level of HA117 RNA expression was significantly higher in the stenotic segment compared to proximal anastomosis (P＜0.001,n=17). RARα were expressed in ganglion cells and nerve fibers between the circular and longitudinal muscles in anastomosis; positive staining was diminished in the stenotic segments. The levels of RARα protein expression was significantly lower in the dilated segment（P＜0.05,n=17）and stenotic segment（P＜0.001, n=17）compared to proximal anastomosis. Conclusion The vitamin A and RARα may contribute to the development of enteric nervous system (ENS); long non-coding RNA (LncRNA) HA117 may have an anti-differentiation function, and play a pivotal role in the progression of HD.
先天性肠无神经节细胞症 / 维生素A / 维甲酸受体α / HA117 / 实时定量聚合酶链反应 / 免疫组织化学 / 免疫印迹法 / 儿童
Hirschsprung’s disease / Vitamin A / Retinoic acid receptor α / / HA117 / / Real-time PCR / Immunohistochemistry / Western blotting / Child
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国家自然科学基金
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