Tumor necrosis factor (TNF) is a pleotropic cytokine involved in a wide range of pathological processes, including inflammatory bowel disease (IBD). In the past, TNF was recognized as a pro-inflammatory cytokine with deleterious effects. This has led to the development of anti-TNF drugs, which revolutionized the treatment of inflammatory disorders such as Crohn's disease. However, in the past 20 years, clinical studies have shown that anti-TNF drugs are not always effective. Moreover, in some rare cases, anti-TNF drugs can even cause an aggravation of the disease. Nowadays, there is increasing evidence that TNF is not only detrimental but can also play an important role in health and the maintenance of homeostasis. The aim of this chapter is to briefly summarize the literature demonstrating the complex dichotomous role of TNF in IBD and discuss the role of anti-TNF drugs in the treatment of IBD.
The choice of appropriate reference genes is essential for correctly interpreting qPCR data and results. However, the majority of animal studies use a single reference gene without any prior evaluation. Therefore, many qPCR results from rodent studies can be misleading, affecting not only reproducibility but also translatability. In this study, the expression stability of reference genes for mRNA and miRNA in archived FFPE samples of 117 C57BL/6JOlaHsd mice (males and females) from 9 colitis experiments (dextran sulfate sodium; DSS) were evaluated and their expression analysis was performed. In addition, we investigated whether normalization reduced/neutralized the influence of inter/intra-experimental factors which we systematically included in the study. Two statistical algorithms (NormFinder and Bestkeeper) were used to determine the stability of reference genes. Multivariate analysis was made to evaluate the influence of normalization with different reference genes on target gene expression in regard to inter/intra-experimental factors. Results show that archived FFPE samples are a reliable source of RNA and imply that the FFPE procedure does not change the ranking of stability of reference genes obtained in fresh tissues. Multivariate analysis showed that the histological picture is an important factor affecting the expression levels of target genes.
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