Verburg Melissa, Renes Ingrid B, Van Nispen Danielle J P M, Ferdinandusse Sacha, Jorritsma Marieke, Büller Hans A, Einerhand Alexandra W C, Dekker Jan
Pediatric Gastroenterology and Nutrition, Department of Pediatrics, Erasmus University, Rotterdam, The Netherlands.
J Histochem Cytochem. 2002 Nov;50(11):1525-36. doi: 10.1177/002215540205001113.
The rapidly dividing small intestinal epithelium is very sensitive to the cytostatic drug methotrexate. We investigated the regulation of epithelial gene expression in rat jejunum during methotrexate-induced damage and regeneration. Ten differentiation markers were localized on tissue sections and quantified at mRNA and protein levels relative to control levels. We analyzed correlations in temporal expression patterns between markers. mRNA expression of enterocyte and goblet cell markers decreased significantly during damage for a specific period. Of these, sucrase-isomaltase (-62%) and CPS (-82%) were correlated. Correlations were also found between lactase (-76%) and SGLT1 (-77%) and between I-FABP (-52%) and L-FABP (-45%). Decreases in GLUT5 (-53%), MUC2 (-43%), and TFF3 (-54%) mRNAs occurred independently of any of the other markers. In contrast, lysozyme mRNA present in Paneth cells increased (+76%). At the protein level, qualitative and quantitative changes were in agreement with mRNA expression, except for Muc2 (+115%) and TFF3 (+81%), which increased significantly during damage, following independent patterns. During regeneration, expression of each marker returned to control levels. The enhanced expression of cytoprotective molecules (Muc2, TFF3, lysozyme) during damage represents maintenance of goblet cell and Paneth cell functions, most likely to protect the epithelium. Decreased expression of enterocyte-specific markers represents decreased enterocyte function, of which fatty acid transporters were least affected.
快速分裂的小肠上皮对细胞生长抑制剂甲氨蝶呤非常敏感。我们研究了甲氨蝶呤诱导损伤和再生过程中大鼠空肠上皮基因表达的调控。在组织切片上定位了10种分化标志物，并相对于对照水平在mRNA和蛋白质水平进行定量。我们分析了标志物之间时间表达模式的相关性。在损伤的特定时期，肠上皮细胞和杯状细胞标志物的mRNA表达显著下降。其中，蔗糖酶-异麦芽糖酶（-62%）和CPS（-82%）呈相关。乳糖酶（-76%）和SGLT1（-77%）之间以及I-FABP（-52%）和L-FABP（-45%）之间也发现了相关性。GLUT5（-53%）、MUC2（-43%）和TFF3（-54%）mRNA的下降独立于任何其他标志物。相反，潘氏细胞中存在的溶菌酶mRNA增加（+76%）。在蛋白质水平，定性和定量变化与mRNA表达一致，除了Muc2（+115%）和TFF3（+81%），它们在损伤期间显著增加，遵循独立模式。在再生过程中，每个标志物的表达恢复到对照水平。损伤期间细胞保护分子（Muc2、TFF3、溶菌酶）表达增强代表杯状细胞和潘氏细胞功能的维持，最有可能是为了保护上皮。肠上皮细胞特异性标志物表达下降代表肠上皮细胞功能下降，其中脂肪酸转运体受影响最小。
