细胞内外环境中凝溶胶蛋白水平对中性粒细胞趋化功能的影响

Effects of extra and intracellular gelsolin levels on polymorphonuclear neutrophil chemotaxis

摘要:

目的：研究细胞内外环境中凝溶胶蛋白水平变化对中性粒细胞（polymorphonuclear neutrophil，PMN）趋化功能的影响。方法：成年SD大鼠，麻醉后采集血液标本，分离PMN，制成1×107个/ml悬液。在Transwell细胞培养系统中进行趋化实验，分别在上、下室培养基中加入重组凝溶胶蛋白，或者采用电穿孔方法向PMN中分别转入凝溶胶蛋白及其抗体，观察PMN趋化功能的变化。结果：在Transwell上、下室加入凝溶胶蛋白，以及向PMN中转入凝溶胶蛋白，趋化至下室的PMN数量和趋化指数均没有明显改变（P ＞0.05）；向PMN中转入凝溶胶蛋白抗体后，趋化至下室的PMN数量减少，趋化指数降低（P＜0.05）。结论：细胞外凝溶胶蛋白水平改变，或向细胞内导入外源性凝溶胶蛋白，均不能影响PMN的趋化能力，但向细胞内导入凝溶胶蛋白抗体，可以抑制PMN的趋化功能。

Abstract:

Objective：To investigate the effects of intracellular or extracellular gelsolin level on N-formylmethionyl-leucyl phenylala－nine（fMLP） induced rat polymorphonuclear neutrophil（PMN） chemotaxis. Methods：PMN neutrophils were isolated from rat venous blood. Freshly prepared PMN suspension was seeded to the upper chamber of Transwell culture system and recombinant gelsolin was added to the upper or lower chamber to investigate the effect of gelsolin on PMN chemotaxis. Recombinant gelsolin or anti-gelsolin antibodies were transfected into PMN with electroporation method and the chemotaxis function was observed respectively. Cell counts in lower chamber were recorded in real time. Results：There was no obvious difference in PMN chemotaxis，regardless of gelsolin was added in the upper or lower chamber or transfected into cytoplasm. However，the rate of chemotaxis and chemotaxis index of PMN electrotransfected with anti-gelsolin antibodies was decreased significantly. Conclusion：Changing intracellular or extracellular gelsolin level exerts on effect on fMLP induced PMN chemotaxis；but electrotransfection with anti-gelsolin antibodies intracellular inhibits fMLP-induced rat PMN chemotaxis.