Hi,
After going through literature studies, I found that in all studies focusing macrophages as target cells for inflammatory disease treatment, the cells were treated with treatment/delivery system (prophylactic) followed by the macrophages activation in vitro using RAW 264.7 cells.
I am wondering why they didn't activate the macrophages first followed by treatment (resembling more to actual disease state) or there is a special reason for treating macrophages first and then activating them?
If you do LPS activation before drug treatment, you are going to end up with high background inflammatory cytokines. This might mask effects of inhibitory drugs, especially for cytokines with long half-lives.
Many of studies aimed to find inhibitors what could prevent activation of signal pathways. If you want to find f.e. inhibitor for iNOS so stimulate cells before adding drug of interest.
@ Petrenyov My objective is not to inhibit signalling pathway. I want to knockdown gene for decreased cytokines production. All literature reports for such cases have activated cells post treatment.
there are two possibilities: one is pre-treatment (frequent in animals rather), other is after-treatment modelling therapeutic window for acute treatments (after disease confirmation). one issue is exists considering macrophage studies: primary cells usually activated during isolation. i think you freely choose your treatment schedule in this case. knockdowning effect has time to develop (and inflammation is disturbing that), this is an other issue in your case. I would try both option to find a more powerful effect for a certain cytokine. some works before, some works after the starts of inflammation. good luck..
If you do LPS activation before drug treatment, you are going to end up with high background inflammatory cytokines. This might mask effects of inhibitory drugs, especially for cytokines with long half-lives.
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