Department of Pediatric Surgery, Children's Hospital of Fudan University, National Children's Medical Center, Shanghai, 201102, PR China.
Department of Pediatric Surgery, Children's Hospital of Fudan University, National Children's Medical Center, Shanghai, 201102, PR China.
J Pediatr Surg. 2023 Jul;58(7):1317-1321. doi: 10.1016/j.jpedsurg.2023.01.062. Epub 2023 Feb 15.
To study the influence of gene methylation in the Shh/Bmp4 signaling pathway on the enteric nervous system in the rectum of rat embryos with anorectal malformations (ARMs).
Pregnant Sprague Dawley rats were divided into three groups; two groups treated with either ethylene thiourea (ETU induce ARM) or ETU+5-azacitidine (5-azaC inhibit DNA methylation) and a normal control group. The levels of DNA methyltransferases (DNMT1, DNMT3a, DNMT3b), the methylation status of the Shh gene promoter region and the expression of the key components were detected by PCR, immunohistochemistry and western blotting.
The expression of DNMTs in the rectal tissue of the ETU and ETU+5-azaC groups was higher than that of the control. The expression of DNMT1, DNMT3a and methylation level of the Shh gene promoter in the ETU group was higher than in the ETU+5-azaC group (P < 0.01). The methylation level of the Shh gene promoter was higher in the ETU+5-azaC group than in the control. The Shh and Bmp4 expression in the ETU and ETU+5-azaC groups were lower than in the control, and their expression in the ETU group was also lower than in the ETU+5-azaC group.
The methylation status of genes in the rectum of the ARM rat model may be changed by intervention. The low methylation level of the Shh gene may promote the expression of key Shh/Bmp4 signaling pathway components.
研究 Shh/Bmp4 信号通路中基因甲基化对肛门直肠畸形（ARM）大鼠胚胎直肠肠神经的影响。
将妊娠 Sprague Dawley 大鼠分为三组；两组用乙硫氨酸脲（ETU 诱导 ARM）或 ETU+5-氮杂胞苷（5-azaC 抑制 DNA 甲基化）处理，一组为正常对照组。通过 PCR、免疫组织化学和 Western blot 检测 DNA 甲基转移酶（DNMT1、DNMT3a、DNMT3b）水平、Shh 基因启动子区的甲基化状态和关键成分的表达。
ETU 和 ETU+5-azaC 组直肠组织中 DNMTs 的表达高于对照组。ETU 组 DNMT1、DNMT3a 的表达及 Shh 基因启动子的甲基化水平均高于 ETU+5-azaC 组（P<0.01）。ETU+5-azaC 组 Shh 基因启动子的甲基化水平高于对照组。ETU 和 ETU+5-azaC 组的 Shh 和 Bmp4 表达均低于对照组，ETU 组的表达也低于 ETU+5-azaC 组。
ARM 大鼠模型直肠基因的甲基化状态可能通过干预发生改变。Shh 基因的低甲基化水平可能促进关键 Shh/Bmp4 信号通路成分的表达。
