Yamada H, Chen D, Monstein H J, Håkanson R
Department of Pharmacology, University of Lund, Sweden.
Biochem Biophys Res Commun. 1997 Feb 24;231(3):835-8. doi: 10.1006/bbrc.1997.6198.
It is generally accepted that while the gene expression of many gut hormones in the pancreas and upper digestive tract is influenced by the prandial state, the expression of house-keeping genes (used as internal standards) is stable. We have analysed how food deprivation affects the messenger (m) RNA expression of gastrin, cholecystokinin (CCK), and somatostatin and of house-keeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-actin and 18S ribosomal (r) RNA. RNA, isolated from oxyntic, antral and duodenal mucosa and pancreas, was subjected to Northern blot analysis using complementary RNA probes. Compared to fed rats, food-deprived rats exhibited reduced mRNA expression of gastrin (antrum), somatostatin (antrum), CCK (duodenum), GAPDH (all tissues studied) and beta-actin (all tissues studied) and unchanged 18S rRNA expression. We conclude that the assessment of gut hormone mRNA expression may be greatly influenced by the choice of internal standard and that 18S rRNA is superior.
人们普遍认为,虽然胰腺和上消化道中许多肠道激素的基因表达受进食状态影响,但管家基因(用作内参)的表达是稳定的。我们分析了禁食如何影响胃泌素、胆囊收缩素(CCK)、生长抑素以及管家基因甘油醛-3-磷酸脱氢酶(GAPDH)、β-肌动蛋白和18S核糖体RNA(rRNA)的信使核糖核酸(mRNA)表达。从胃底、胃窦、十二指肠黏膜和胰腺分离的RNA,使用互补RNA探针进行Northern印迹分析。与喂食的大鼠相比,禁食大鼠的胃泌素(胃窦)、生长抑素(胃窦)、CCK(十二指肠)、GAPDH(所有研究组织)和β-肌动蛋白(所有研究组织)的mRNA表达降低,而18S rRNA表达未改变。我们得出结论,肠道激素mRNA表达的评估可能受内参选择的极大影响,且18S rRNA更具优势。