同源盒基因／骨形态发生蛋白-4信号在小儿先天性肛门直肠畸形中的表达  

The expression and role of homebox/bone morphogenetic protein 4 signal in children with congeni- tal anorectal malformations

作　　者：周良[1] 侯广军[1] 耿宪杰[1] 张现伟[1] 田亮[2] 

机构地区：[1]郑州市儿童医院普外科,450018 [2]郑州市儿童医院血液肿瘤科,450018

出　　处：《中华实验外科杂志》2017年第2期240-242,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金-河南人才培养联合基金（U1204806）

摘　　要：目的探讨同源盒基因（Hox）/骨形态发生蛋白-4（BMP-4）信号在小儿先天性肛门直肠畸形中的表达。 方法实时定量反转录聚合酶链反应（RT-qPCR）检测音猬因子（Shh）、BMP-4和Hox在小儿先天性直肠肛门畸形中的表达水平。用乙烯硫脲诱导孕鼠胎仔产生先天性肛门畸形鼠胚。免疫组织化学和RT-qPCR检测受孕后第12、13、14、15、16天胎仔直肠末端Shh、BMP-4和Hox的表达。Western blot检测受孕第16天胎仔直肠末端Shh、BMP-4和Hox水平。 结果对照组中Shh、BMP-4和Hox水平约为小儿先天性肛门直肠高位畸形组织中2倍；低位畸形组织中Shh、BMP-4和Hox水平与对照组比较没有明显变化。免疫组织化学结果发现，对照组和乙烯硫脲组中Shh、BMP-4和Hox在13 d平均吸光度值达到高峰，在14、15、16 d出现明显下降；对照组和乙烯硫脲组Shh 13 d时平均吸光度值依次为：0.405±0.014、0.284±0.024，BMP-4 13 d时平均吸光度值依次为：0.287±0.023、0.214±0.012 ，Hox 13 d时平均吸光度值依次为：0.395±0.029、0.199±0.015，Shh、BMP-4和Hox水平均低于对照组。对照组和乙烯硫脲组中BMP-4 mRNA水平在13 d有所增高，从14 d开始出现明显下降。对照组和乙烯硫脲组中Shh、Hox mRNA水平在12、13、14 d表达较高，而15 d表达水平明显下降，16 d有微弱回升。乙烯硫脲组中Shh、BMP-4和Hox蛋白表达水平均明显低于对照组，Shh从1.314±0.024下降至1.035±0.018，BMP-4从1.412±0.034下降至0.854±0.028，Hox从1.052±0.014下降至0.632±0.041。 结论Hox/BMP-4信号在小儿先天性肛门直肠高位畸形中低表达，乙烯硫脲可以干扰胚胎发育过程中Hox/BMP-4信号表达。Objective To investigate the expression of homebox （Hox）/bone morphogenetic pro- tein 4 （ BMP - 4） signal in children with congenital anorectal malformations. Methods Real - time quan- titative reverse transcriptase - polymerase chain reaction （ RT - qPCR） was used to detect expression levels of Sonic Hegehdog, Bone morphogenetie protein 4 and homebox gene in children with congenital anorectal malformations. The use of ethylene thiourea to induce fetal congenital malformation of the fetus in pregnant mice. After fertilization 12, 13, 14, 15, 16 d, Shh, BMP-4 and Hox were detected by immunohisto- chemistry and RT - qPCR. The expression of Shh, BMP -4 and Hox were detected after fertilization 12 d by Western blotting. Results The levels of Shh, BMP - 4 and Hox in the control group were about 2 times higher than that in the children with congenital anorectal malformation. The levels of Shh, BMP -4 and Hox were not significantly different in the low deformity compared with the control group. The results of im- munohistochemistry showed that Shh, BMP - 4 and Hox in the control group and the ethylene thiourea group reached peak values of 13 d average absorbance value, In 14, 15, 16 d decreased significantly; the average absorbance value of the control group and the ethylene thiourea group Shh of 13 d were： 0. 405 ± 0. 014, 0. 284±0. 024, the average absorbance value of BMP -4 of 13 d were： 0. 287 ±0. 023,0. 214 ± 0. 012, the average absorbance value of Hox of 13 d were ： 0. 395 ±0. 029, 0. 199 ± 0. 015, ethylene thiou- rea group in Shh and BMP -4 and Hox level were lower than control group. BMP -4 mRNA levels in the control group and the ethylene thiourea group were significantly decreased at 13 d, and the 14 d level was significantly decreased. The 14, 12 d Shh, Hox mRNA levels were higher in the control group and the eth- ylene thiourea group, while the expression level of 15 d was significantly decreased, and the 16 d level was slightly higher. The expression levels of Shh, BMP - 4 and Hox we

关 键 词：先天性 肛门 直肠 畸形 同源盒基因/骨形态发生蛋白-4信号 

分 类 号：R725.7[医药卫生—儿科]