We have determined the expression of TNF-a, IL-1b, and IL-6 mRNA in MCF-7 and RAW264.7 cells after heat exposure and Iscador treatment. I need some information for discussion, how to interpret these results? I mean, is better when the expression in falling down or rather when is going up?
Hey Henryk,
it depends on the context. People observed up-regulation of these three prominent pro-inflammatory cytokines in macrophages after many different stimuli (PAMPs, pathogen infection, stress, other cytokines and so on).
In your in vitro setting with a macrophage-like cell line there is no further interpretation of the results. You treated the cells and mRNAs go up. The next steps could be:
1. Checking your results with primary macrophages (BMDM or tissue-resident macrophages).
2. Checking if your cytokines are really produced and secreted, for example with an commercially available ELISA (we observed many times that just the presence of mRNA does not necessarly mean that the cytokines are produced and secreted).
3. Checking an in vivo setting (putting your pro-inflammatory response in a setting where it could have an important impact, which is not the case in a culture well).
I hope that helps a little.
All the best and a nice weekend.
Marc
Hi Henryk,
Briefly, these are pro-inflammatory cytokines, so if your treatments (the heat exposure and Iscador treatment) caused an upregulation of these genes, this means that the treatments have an inflammatory effect, but in order to say that you need to measure the protein secretion, because the genes are not always expressed into proteins. You can measure the protein secretion using ELISA kit. However, if your treatments caused a down regulation in the expression of these genes (plus decrease in protein secretion), then your treatments have an anti-inflammatory effect.
I hope this helps!
All the best,
Alaa.