Chen H, Tian W, Lei S, Wei Y, Chen Y
Institute of Biomedical Engineering, West China University of Medical Sciences, Chengdu 610041.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 1999 Sep;16(3):288-94.
Fluid shear is a physiological and functional environment for circulating neutrophils where various chemoattractants and cytokines have different effects on neutrophils, activating them or evoking inflammation. So far there have been abundant in vitro researches demonstrating that while neutrophils are activated, some adhesion molecules, which are expressed on the surface of neutrophils, are necessary to cell-to-cell adhesion, and the quantity or quality of these molecules will be changed. The above results were all obtained under static fluid which is quite different from physiological condition. Recently, we used Low-Shear 30 to provide steady or sinusoidal-oscillated fluid shear stress on separated neutrophils with or without f-MLP/TNF stimulation. FITC-anti CD18 monoclonal antibody and PE-anti CD62L monoclonal antibody were added to label CD18 and CD62L on neutrophils suface respectively. Flow cytometry was used to quantify surface expression and positive cell percentage of these two adhesion molecules. The results demonstrat that fluid shear stress increases the expression of CD18 significantly and decreases the CD62L positive cell percentage, but it does not influence the CD62L expression on the CD62L positive cells. The effects of the different patterns of fluid shear stress are not much different, f-MLP or TNF can slightly decrease the expression of CD18 and make CD62L shedding respectively. When one of the two activators and the fluid shear stress act together, the expression of the two adherent molecules can be added. This indicates that the mechanical and biochemical stiumlative action can produce different effects through different receptors and different signal transduction pathways. So the mechanical environment should be considered in the research of activation and adhesion of neutrophils.
流体剪切力是循环中性粒细胞所处的一种生理和功能环境,在此环境中,各种趋化因子和细胞因子对中性粒细胞有不同影响,可激活它们或引发炎症。到目前为止,已有大量体外研究表明,中性粒细胞被激活时,其表面表达的一些黏附分子对细胞间黏附是必需的,且这些分子的数量或质量会发生变化。上述结果均是在与生理状况差异很大的静态流体条件下获得的。最近,我们使用低剪切力装置30为分离的中性粒细胞提供稳定的或正弦振荡的流体剪切应力,同时给予或不给予f-MLP/TNF刺激。分别加入异硫氰酸荧光素(FITC)标记的抗CD18单克隆抗体和藻红蛋白(PE)标记的抗CD62L单克隆抗体,以标记中性粒细胞表面的CD18和CD62L。采用流式细胞术对这两种黏附分子的表面表达及阳性细胞百分比进行定量分析。结果表明,流体剪切应力显著增加CD18的表达,降低CD62L阳性细胞百分比,但不影响CD62L阳性细胞上CD62L的表达。不同模式的流体剪切应力的影响差异不大,f-MLP或TNF可分别轻微降低CD18的表达并使CD62L脱落。当两种激活剂之一与流体剪切应力共同作用时,两种黏附分子的表达可叠加。这表明机械和生化刺激作用可通过不同受体和不同信号转导途径产生不同效应。因此,在中性粒细胞激活和黏附的研究中应考虑机械环境因素。