RAW 264.7 cells exhibit high response to LPS even in the low concentration of 10ng/ml. And the nitric oxide in the supernatant of cells cocultured with LPS (10ng/ml) after 24H would rise to 60uM sometimes even to 90uM, although the control without LPS stimulation was about 5uM. And even the inhibitor BAY 11-7082 shows little suppression on the nitric oxide in the cells cultured medium induced by LPS.
Why do RAW cells become so sensitive to LPS stimulation? Is it normal?
RAW cells treated with LPS or IFN Gamma differentiate into M1 subtype. Because M1 cells are cosidered to be proinflammatory they release lot of inflammatory substances such as TNF Alpha, NO, SO etc., Small vacuoles can also be noticed in RAW (as a matter of fact any other macrophage cell line) cells treated with either LPS of IFN Gamma. Due to these inflammatory substances produced, the cells turn very fragile and show decreased viability. May be you need to consider TNF Alpha neutralization too along with NO inhibitors.
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