[Effects of IL-8 on neutrophils under fluid shear]

In order to elucidate the effect of IL-8 on neutrophils in a systematic and all-round way, we introduced an important physiological environment of neutrophils, fluid flow, into this study on the changes in some responses of neutrophils to IL-8 and the inter relationship of those responses under the conditions of different patterns of flow field.

After using IL-8 and steady flow or sinusoidal-oscillate fluid shear rate simultaneously acting on separated neutrophils for one minute, we measured the expression of CD18, CD62L and the F-actin content with flow cytometry and measured the free calcium concentration inside cells ([Ca2+]i) with the fluorescent stain Fura2/AM using the fluorescent spectrophotometer.

It was found that flow shear stress could influence the expression of surface adherence molecule of neutrophils under the action of IL-8. CD18 surface expression increased, and CD62L shed under different flow shear rates, but no significant relationship of these changes with the intensity and pattern of shear stress was noted. On the other hand, the F-actin content in neutrophils under the action of shear stress and IL-8 decreased significantly at low shear rate and then gradually increased at high shear rate (600s -1) to the control value (without shear). The [Ca2+]i concentration under the action of shear stress and IL-8 decreased significantly at low shear rate followed by a quick increase at high shear rate.

The flow shear stress can influence the activation effects of IL-8 on neutrophils and regulate the magnitude of neutrophil reactions to a great extent, and among these reactions, the change of [Ca2+]i concentration is the most sensitive one and the greatest one. This is in consistence with its second messenger role in the signal transduction pathway. Our results could serve as a basis on which to understand better the mechanism of inflammatory reaction wherein the neutrophils under flow condition of blood are involved.