Abstract
In rat neutrophils, formylmethionyl-leucyl-phenylalanine (fMLP)-induced inositol phosphate formation was concentration-dependently inhibited by acetylshikonin as well as by a putative phospholipase C (PLC) inhibitor [6-[[17beta-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-p yrrole-2,5-dione (U73122). The IC50 value of acetylshikonin for the inhibition of inositol trisphosphate (IP3) formation was estimated to be 16.1 +/- 1.5 microM. The reduction of inositol phosphate levels appeared to reflect inhibition of PLC activity because the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) catalyzed by a soluble fraction from neutrophils was also inhibited by acetylshikonin (IC50 value 21.4 +/- 6.1 microM) over the same range of concentrations. Although acetylshikonin alone evoked Ca2+ and Mn2+ influx into neutrophils in Ca2+-containing medium, acetylshikonin, like U73122, inhibited Ca2+ release (IC50 value approximately 5.3 +/- 0.4 microM) from internal stores in Ca2+-free medium. These results indicate that acetylshikonin inhibits phosphatidylinositol signaling in neutrophils.
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