Upregulation of PPPDE1 contributes to anorectal malformations via the mitochondrial apoptosis pathway during hindgut development in rats
Upregulation of PPPDE1 contributes to anorectal malformations via the mitochondrial apoptosis pathway during hindgut development in rats
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大鼠后肠发育过程中PPPDE1的上调通过线粒体凋亡途径导致肛门直肠畸形
DOI：
10.1016/j.yexcr.2021.112574
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发表时间：
2021-04-01
影响因子：
3.7
通讯作者：
Bai，Yu Zuo
中科院分区：
文献类型：
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作者：
Li，Si Ying;Wang，Chen Yi;Bai，Yu Zuo
先天性肛门直肠畸形（ARMs）是胃肠道最突出的畸形之一，然而，其确切的病因仍然不明。免疫组化结果显示，在ARM组中，PPPDE 1阳性细胞广泛分布于GD 13至GD 16的后肠上皮组织中。免疫荧光显示，大多数TUNEL-，Bax-和细胞色素C（Cyt C）-阳性细胞重叠PPPDE 1阳性细胞在尿直肠隔（URS）。Western blotting和实时定量RT-PCR显示，从GD 13到GD 14，ARM组中PPPDE 1水平显著较高（p< 0.05）。用PPPDE 1过表达质粒/NC（阴性对照）或si-PPPDE 1/si-NC转染IEC-6细胞。流式细胞术分析和CCK-8测定（分别用于检测凋亡和增殖）以及蛋白质印迹法显示PPPDE 1的水平与促凋亡分子Bax和Cyt C正相关。因此，PPPDE 1异常高表达导致了ARMs胎鼠后肠发育过程中的时空不平衡。因此，PPPDE 1的上调可能通过线粒体凋亡途径促进上皮细胞凋亡并减少后肠的增殖。这可能会影响URS和泄殖腔膜的融合，最终抑制后肠发育并导致ARM。
Congenital anorectal malformations (ARMs) are among the most prominent deformities of the gastrointestinal tract; however, their precise aetiology remains obscure. Immunohistochemistry demonstrated that, in the ARM group, the PPPDE1-positive cells were widely distributed in the hindgut epithelial tissue from GD13 to GD16. Immunofluorescence revealed that most TUNEL-, Bax-, and Cytochrome C (Cyt C)-positive cells overlapped with PPPDE1-positive cells in the urorectal septum (URS). Western blotting and quantitative real-time RT-PCR revealed that PPPDE1 levels were significantly higher in the ARM group from GD13 to GD14 (p< 0.05). IEC-6 cells were transfected with PPPDE1 overexpression plasmid/NC (negative control) or si-PPPDE1/si-NC. Flow cytometry analysis and CCK-8 assay (used to detect apoptosis and proliferation, respectively), as well as western blotting, showed that the levels of PPPDE1 were positively correlated with the pro-apoptotic molecules Bax and Cyt C. Accordingly, aberrantly high expression of PPPDE1 caused a spatiotemporal imbalance in foetal rats with ARMs during hindgut development. Therefore, the upregulation of PPPDE1 may promote epithelial apoptosis and reduce proliferation in the hindgut via the mitochondrial apoptotic pathway. This could affect the fusion of the URS and cloacal membrane, ultimately inhibiting the hindgut development and resulting in ARMs.