Sox4基因在小鼠腭突发育与腭裂形成过程中的表达及作用

发布时间：2018-05-19 03:27

本文选题：Sox4 + 腭裂　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:探讨Sox4基因在BALB/c正常小鼠与腭裂模型腭突发育过程中的表达差异,初步研究该基因在小鼠上腭发育过程中的作用及其与腭裂发生的关系。方法:腭裂发生率统计:将12只BALB/c孕鼠平均分为两组,将实验组孕鼠于GD10(gestation day 10)一次性管饲50mg/kg维甲酸(retinoic acid,RA),对照组孕鼠给予管饲等量玉米油。在GD17上午8时将孕鼠处死,剖腹取得胎鼠,放大镜下观察每组胎鼠腭裂形成情况,统计腭裂发生率。Sox4蛋白检测:按照上述方法获取对照组及实验组孕鼠各15只。分别在GD13、GD14、GD15上午8时处死实验组与对照组孕鼠各5只,剖腹剪取胎鼠头部标本共253例,常规石蜡包埋及切片。利用HE染色观察胎鼠腭突发育过程中的形态变化,采用免疫组织化学方法检测目的蛋白在各组间不同时期腭突中的表达差异。结果:1.在统计腭裂发生率的实验中对照组共获得胎鼠55只,其中腭裂胎鼠为0只;实验组共获得胎鼠52只,腭裂胎鼠为44只。由此得出GD10给予50mg/kg维甲酸诱导BALB/c小鼠腭裂发生率为84.6%.可诱导稳定的腭裂模型。2.获得HE染色切片23例,结果显示对照组双侧腭突在GD14发生水平向翻转并接触融合,实验组腭突翻转推迟至GD15但并未融合。3.免疫组化实验共获得切片219例,结果显示,Sox4在实验组与对照组腭突的被覆上皮及腭中线上皮中均有表达,用平均光密度值[1]分析对照组GD13-GD15组内两两对比皆有差异(p0.05),且在GD14表达量最高。实验组中GD13、GD14与GD15之间的表达对比均无差异(p0.05)。实验组与对照组组间对比在GD13表达无差异(p0.05),GD14对照组高于实验组(p0.05),GD15实验组高于对照组(p0.05)。结论:Sox4在腭裂与正常腭突中GD14-GD15的表达存在明显差异,在腭突融合期发挥调节作用。
[Abstract]:Objective: to investigate the difference of expression of Sox4 gene in the development of palatal process in normal BALB/c mice and cleft palate model, and to study the role of Sox4 gene in the development of upper palate in mice and its relationship with cleft palate. Methods: the incidence of cleft palate in 12 pregnant BALB/c rats was divided into two groups on average. The pregnant rats in the experimental group were given 50mg/kg retinoic acididate once, and the pregnant rats in the control group were given the same amount of corn oil. The pregnant mice were killed at 8: 00 am in GD17, and the fetal rats were obtained by laparotomy. The cleft palate was observed in each group under a magnifying glass, and the incidence rate of cleft palate. Sox4 protein was detected. According to the above methods, 15 pregnant rats in the control group and 15 in the experimental group were obtained. Five pregnant rats of the experimental group and the control group were killed at 8: 00 am of GD13, GD14 and GD15, and 253 fetal rat head specimens were taken by caesarean section. The specimens were embedded and sliced in paraffin. The morphological changes of palatine process in fetal rats were observed by HE staining and the expression of target protein in palatine process at different stages was detected by immunohistochemical method. The result is 1: 1. In the experiment of counting the incidence of cleft palate, 55 fetal rats were obtained in the control group, including 0 in the experimental group, 52 in the experimental group and 44 in the cleft palate fetus. The incidence of cleft palate in BALB/c mice induced by 50mg/kg retinoic acid by GD10 was 84. 6%. It can induce stable cleft palate model. He stained sections were obtained in 23 cases. The results showed that the bilateral palatine process in the control group was horizontally reversed and fused in contact with GD14, while the palatine process turnover in the experimental group was delayed to GD15 but did not fuse. 3. A total of 219 specimens were obtained by immunohistochemical method. The results showed that Sox4 was expressed in the coated epithelium and midline epithelium of palatine process in the experimental group and the control group. The average optical density value [1] was used to analyze the difference between two contrast groups in the GD13-GD15 group of the control group, and the highest expression of GD14 was found in the control group. There was no difference between the expression of GD13, GD14 and GD15 in the experimental group (p0.05). There was no difference in the expression of GD13 between the experimental group and the control group. The expression of GD14 in the control group was higher than that in the experimental group (p0.05) and that in the control group was higher than that in the control group (P 0.05). Conclusion the expression of GD14-GD15 in cleft palate is significantly different from that in normal palatine process, and it plays a regulatory role in palatine process fusion.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R782.2

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