BMP7 is Downregulated in Lumbosacral Spinal Cord of Rat Embryos With Anorectal Malformation
BMP7 is Downregulated in Lumbosacral Spinal Cord of Rat Embryos With Anorectal Malformation
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BMP7 在肛门直肠畸形大鼠胚胎腰骶脊髓中表达下调
DOI：
10.1016/j.jss.2019.11.020
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发表时间：
2020-07-01
影响因子：
2.2
通讯作者：
Wang，Weilin
中科院分区：
文献类型：
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作者：
Yang，Zhonghua;Li，Xiang;Wang，Weilin
骨形态发生蛋白(BMP)是一个高度保守的信号蛋白家族，参与脊髓的形成、发育和分化。腰椎脊髓畸形与直肠肛管畸形(ARM)术后并发症有关。然而，这些畸形发生的机制仍然不清楚。材料和方法采用乙硫脲(ETU)诱导的胚胎大鼠手臂模型，研究BMP7在腰骶段脊髓中的表达。分别于胚胎16、17、19、21天用免疫组织化学染色、qRT-PCR和Western印迹分析Bmp7的表达。结果BMP7mRNA(E16，1.041±0.169比0.758±0.0.0423，P<；0.05；E17，1.889±0.444vsus1.601±0.0.263，P<；0.05；E19，2.898±0.434比1.981±0.068，P<；0.01；E21，2.652比0.637比1.957±0.09，P<；0.05；和蛋白质(E16，1.068±0.065对0.828±0.066，P<；0.01；E17，1.728±0.153对1.4±0.148，P<；0.05；E19，2.313±0.141对1.696±0.21，P<；0.01；E21，2.021±0.13对1.43±0.128，P<；0.01；(0.01)表达下调，在有臂胚胎中，位于腰骶髓背角的中间神经元(IN)的表达水平特别低。在胚胎19天，Western印迹分析显示P-Smad1/5降低(1.13±0.08比0.525±0.06，P<；0.01)。结论BMP7表达下调可能与胚胎发育过程中脊髓发育不良有关，提示BMP7可能在ARM的发病机制及其并发症中起重要作用。
BackgroundBone morphogenetic proteins (BMPs) comprise a highly conserved signaling protein family, which are involved in spinal cord formation, development and differentiation. Malformations of the lumbosacral spinal cord are associated with postoperation complications of anorectal malformation (ARM). However, the mechanism underlying the development of these malformations remains elusive.Materials and methodsEmbryonic rat ARM model induced by ethylenethiourea (ETU) was introduced to investigate BMP7 expression in lumbosacral spinal cord. BMP7 expression was analyzed by immunohistochemical staining, qRT-PCR, and Western blot analysis on embryonic (E) days 16, 17, 19, and 21. The expression of the neuronal marker neurofilament (NF) and pSmad1/5 was determined by immunofluorescence double staining and Western blot analysis during peak BMP7 expression.ResultsBMP7 mRNA (E16, 1.041 ± 0.169versus0.758 ± 0.0423,P< 0.05; E17, 1.889 ± 0.444versus1.601 ± 0.263,P< 0.05; E19, 2.898 ± 0.434versus1.981 ± 0.068,P< 0.01; and E21, 2.652 ± 0.637versus1.957 ± 0.09,P< 0.05;) and protein (E16, 1.068 ± 0.065versus0.828 ± 0.066,P< 0.01; E17, 1.728 ± 0.153versus1.4 ± 0.148,P< 0.05; E19, 2.313 ± 0.141versus1.696 ± 0.21,P< 0.01; and E21, 2.021 ± 0.13versus1.43 ± 0.128,P< 0.01) were downregulated, and their expressions were specifically low in interneurons (IN) located in the dorsal horn of the lumbosacral spinal cord in embryos with ARM. On E19, Western blot analysis revealed reduced P-Smad1/5(1.13 ± 0.08versus0.525 ± 0.06,P< 0.01).ConclusionsAn implication of this study is the possibility that BMP7 downregulation contributes to maldevelopment of the lumbosacral spinal cord during embryogenesis in fetal rats with ARM, indicating that BMP7 may play an important role in ARM pathogenesis and the complications thereof.