[Changes of F-actin in neutrophils under fluid shear stress]

Neutrophils are the main defender in the body, and their physiological characters are associated with functions. Because the environments of neutrophils maturation and storage are different from their function environment, they have to adapt thess changes. Filamentous actin (F-actin), as one of the important cytoskeleton components of neutrophils, has different quantity and distribution under different physiological conditions. Previous researches are all focused on static activation of neutrophils using various stimulants such as formyl-methionyl-leucyl-phenylalanine (f-MLP) and tumor necrosis factor(TNF). In our study, Low-Shear 30 and NXE-1 rheometers were used to provide steady or sinusoidaloscillatory fluid shear stress on separated neutrophils. Fluorescent agent Tritc-Phalloidin was added to label F-actin and the mean fluorescent intensity tested by flow cytometry was used as the indication of F-actin quantity and confocal laser scan microscope was used to detect the distribution of F-actin in neutrophils. We found that F-actin polymerization was significantly decreased under both steady and sinusoidal-oscillatory shear stress when the shear stress was below certain level. The density of the cortex F-actin near cell membrane in cells sheared became thinner than in that without shear stress. f-MLP and TNF both could increase actin polymerization in neutophils due to activation. When neutorphils were activated with f-MLP or TNF under shear stress, the quantity of F-actin in the cells was also decreased, but it was still higher than that in the cells absent of f-MLP or TNF. We concluded that neutrophils had different mechanical properties in different fluid environments. F-actin content and distribution would change according to the shear stress values to play their determinant role in neutrophils mechanical adaptation. The mechanism of actin polymerization in neutrophils for mechanical action is probably different from that for chemoattractants and cytokine activation.