摘要
采用重复密度梯度离心的方法分离中性粒细胞,并对其在10～100 nmol/L fMLP不同浓度趋化诱导下产生极性化的极性化率和伪足长度变化趋势进行分析。结果表明在加入fMLP的短时间内,细胞都表现出极性化率明显上升的趋势,以100 nmol/L为最快,但在3～4 min内都会达到90%以上的极性化水平。同时,细胞伪足长度明显受到不同fMLP浓度刺激的影响,表现为伸展和回缩相的交替,从而组成一个振荡周期。组成振荡周期的伸展相和回缩相及振荡频率都明显受到不同fMLP浓度刺激的影响,依据伪足变化率本文提出了细胞极性活跃程度的分类。由于伪足的变化与F-actin的聚合密切相关,而F-actin的聚合又受到极性信号分子的调节,因此对伪足变化的分析有助于了解中性粒细胞极性信号传导通路的调节机制。
To analyze the polarization rate and the changes of pseudopod length in neutrophil, Percoll density gradient centrifugation upon the stimulation of 10^-7-10^-8 mol/L N-formyl-methionyl-leucyl-phenylalanine （fMLP） was used. The polarization rate of neutrophil was significantly increased after treated by 10 nmol/L, 50 nmol/L and 100 nmol/L fMLP separately in 4min, and reached 90% from 3 min to 4 min in all three groups. There was fastest increase of the polarization rate in the group of 100 nmol/L fMLP treatment. The pseudopod length of neutrophil was affected markedly by stimulation of the different concentrations of fMLP, which showed an oscillatory phenomenon of the pseudopod extention and retraction and an oscillatory cycle was formed. The oscillatory frequency and the oscillatory cycles were significantly affected by stimulation of the different concentration of fMLP. The neutrophils were classified by their activity of polarization in this study according to the change rate of peseudopod. The change of pseudopod was represented by F-actin polymerization which was modulated by polarization signaling molecule. The analysis of the pseudopod change could contribute to the knowledge of the signaling mechanism in neutrophil polarization.
出处
《中国细胞生物学学报》
CAS
CSCD
2010年第4期583-588,共6页
Chinese Journal of Cell Biology
基金
国家自然科学基金(No.30971193
No.30800438
No.30672353)资助项目~~
