摘要
目的探讨胎鼠后肠发育与sonichedgehog(Shh)基因转录因子Gli2基因表达水平的关系。方法48只妊娠Wistar大鼠随机分成2组,实验组(n=24)在孕11d按125mg/kg胃管注入1%乙烯硫脲(ethylenethiourea,ETU),对照组(n=24)给予等量蒸馏水。两组分别于孕12d、14d和16d各处死8只母鼠,每只母鼠取4只体重相似的胎鼠,取其直肠1cm提取RNA,采用半定量RT_PCR法检测Gli2基因表达。结果实验组胎鼠直肠Gli2基因表达水平明显低于对照组(P<0.05),实验组Gli2基因表达呈一低水平线,而对照组随妊娠时间呈一下降的曲线。结论Gli2基因表达水平在胎鼠后肠发育中具有重要的作用。
Objective It is recently reported a possible role of Sonic hedgehog(Shh)gene and its protein receptor Gli2,Gli3 in the pathogenesis of anorectal malformations. The study tried to explore the relationship between the Shh signaling molecule system and the hindgut development of rat embryos. Methods Seventy-two healthy non-pregnant Wistar rats weighing 220-250 g(male 48 and female 24)kept at a cage overnight at a ratio of 2 female to 1 male rat.A vagina swab was made in female rat the next morning.Mating was suggested if sperms were observed at light microscope and the very day was designated as gestational day. Forth-eight pregnant Wistar rats were divided randomly into control(n=24)and experimental group (n=24).The experimental rats received 1% ETU(125 mg/kg)by gavage on gestational day 11,and the control rats received the same amount of distilled water and then continued their pregnancy.Every eight pregnant rats were killed on gestational day 12,14 and 16 in each group.The fetuses were taken out by caesarean section under sterile condition.It was obtained cm of rectum samples of each 4 similar-weight fetuses who came from each pregnant rat.The specimens were then put into the sterile DEPC(diethylpyrocarbonate)-treated eppendorf tube for liquid nitrogen storage.RNA extraction was then performed.PCR products were further examined by one-step PCR method.The Gli2/β-tubulin relative density value ratio was employed to reflect the level of Gli2 gene expression. Results The Gli2 gene expression in experimental group was lower than that in control group (P<0.05).The level of Gli2 gene expression in normal embryo rectum didn't show time-dependent change and kept at a low level,while the level of Gli2 gene expression in ETU-treated group decreased over time,being a declined curve along the pregnant time with the highest level at 12 d and the lowest level at 16 d. Conclusions The level of Gli2 gene expression is likely to be responsible for the formation of the rat fetus hindgut development.
出处
《临床儿科杂志》
CAS
CSCD
北大核心
2005年第7期477-480,共4页
Journal of Clinical Pediatrics
