论文部分内容阅读
目的为研究alk-SMase在结肠癌发生发展中的作用提供动物模型的实验数据。方法将野生型(WT,+/+)、杂合子(+/-)、纯合子(KO,-/-)alk-SMase基因剔除小鼠剪尾提取DNA,PCR法作基因型鉴定;取其肝肠组织进行HE染色、激光共聚焦分析以及质谱仪检测,观察和分析不同基因型鼠的表型特征。结果与WT和杂合子比较,KO鼠显示:外观和体重无明显区别;基因型鉴定出现一个条带,其分子量为247 bp;小肠黏膜明显增厚,但alk-SMase表达减弱;质谱分析显示在肠道和肝脏中的鞘磷脂含量增高,1-磷酸鞘氨醇含量增加,而神经酰胺含量降低。结论 alk-SMase KO鼠有明显特征,但其外观和体重与其他基因型比较没有明显区别,为研究alk-SMase的生物功能提供了一个理想的动物模型。
Objective To provide experimental data for studying the role of alk-SMase in the development of colon cancer. Methods Wild-type (WT, + / +), heterozygous (+/-) and homozygous (KO, - / -) alk-SMase genes were excised from the tail of a mouse to extract DNA. PCR was used to identify the genotype. Hematoxylin-eosin staining, confocal laser scanning microscopy and mass spectrometry were used to detect and analyze the phenotypic characteristics of different genotype mice. Results Compared with WT and heterozygotes, KO mice showed no significant difference in appearance and body weight. A band with a molecular weight of 247 bp was found in the genotype. The mucosa of the small intestine was thickened but the expression of alk-SMase was weakened. The results of mass spectrometry Sphingomyelin levels in the gut and liver increased, sphingosine-1 -phosphate increased, and ceramide decreased. Conclusions The alk-SMase KO mouse has obvious characteristics, but its appearance and weight are not significantly different from other genotypes, which provides an ideal animal model for studying the biological function of alk-SMase.