乙基硫脲诱导的肛直肠畸形大鼠胚胎肛直肠发育过程中 TBX4 的异常表达。
Abnormal expression of TBX4 during anorectal development in rat embryos with ethylenethiourea-induced anorectal malformations.
机构信息
Department of Pediatric Surgery, Shengjing Hospital of China Medical University, No. 36 SanHao St., Heping District, Shenyang, 110004, China.
出版信息
Biol Res. 2019 May 4;52(1):27. doi: 10.1186/s40659-019-0235-6.
BACKGROUND
To assess the expression of T-box transcription factor 4 (TBX4) during the anorectal development in normal and ethylenethiourea (ETU)-induced anorectal malformations (ARM) rat embryos.
METHODS
Anorectal malformations was induced by ETU on the 10th gestational day (E10) in rat embryos. Spatio-temporal expression of TBX4 was evaluated in normal (n = 490) and ETU-induced ARM rat embryos (n = 455) from E13 to E16 by immunohistochemical staining, Western blot analysis and real-time RT-PCR.
RESULTS
In the normal embryos, immunohistochemical staining revealed that TBX4 expression was detected in the epithelium of hindgut and urorectal septum (URS) on E13. TBX4-immunopositive cells were increased significantly in the epithelium of hindgut and URS, the future anal orifice part of cloacal membrane on E14. On E15, abundant stained cells were observed in the rectum, URS and dorsal cloacal membrane and the expression of positive cells reached its peak. On E16, only sporadic positive cells were distributed in the epithelium of the distal rectum. In the ARM embryos, the hindgut/rectum, URS and dorsal cloacal membrane were faint for TBX4 immunohistochemical staining. In the normal group, TBX4 protein and mRNA expression showed time-dependent changes in the hindgut/rectum from E13 to E16 on Western blot and real-time RT-PCR. On E13 and E15, the expression level of TBX4 mRNA in the ARM group was significantly lower than that in the normal group (P < 0.05). On E15, the expression level of TBX4 protein in the ARM group was significantly lower than that in the normal group (P < 0.05).
CONCLUSIONS
The expression of TBX4 was downregulated in ETU-induced ARM embryos, which may play important roles in the pathogenesis of anorectal development.
背景
评估 T 盒转录因子 4（TBX4）在正常和乙撑硫脲（ETU）诱导的肛门直肠畸形（ARM）大鼠胚胎肛门直肠发育过程中的表达。
方法
在大鼠胚胎第 10 天（E10）用 ETU 诱导肛门直肠畸形。通过免疫组织化学染色、Western blot 分析和实时 RT-PCR 评估正常（n=490）和 ETU 诱导的 ARM 大鼠胚胎（n=455）在 E13 至 E16 期间 TBX4 的时空表达。
结果
在正常胚胎中，免疫组织化学染色显示 TBX4 表达于 E13 时的后肠和尿直肠隔（URS）上皮。E14 时，后肠和 URS 上皮、肛膜未来的肛门口部分的 TBX4 免疫阳性细胞明显增加。E15 时，在直肠、URS 和背侧肛膜中观察到大量染色细胞，阳性细胞表达达到高峰。E16 时，仅在远端直肠的上皮中分布有散在的阳性细胞。在 ARM 胚胎中，后肠/直肠、URS 和背侧肛膜的 TBX4 免疫染色较弱。在正常组中，Western blot 和实时 RT-PCR 显示 TBX4 蛋白和 mRNA 表达在 E13 至 E16 期间在后肠/直肠中呈时间依赖性变化。在 E13 和 E15 时，ARM 组的 TBX4 mRNA 表达水平明显低于正常组（P<0.05）。在 E15 时，ARM 组的 TBX4 蛋白表达水平明显低于正常组（P<0.05）。
结论
ETU 诱导的 ARM 胚胎中 TBX4 的表达下调，可能在肛门直肠发育的发病机制中发挥重要作用。
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