C5a对中性粒细胞和肾小球内皮细胞粘附行为的影响

The effect of C5a on properties of adhesion between granulocytes and glomerulus vascular endothelial cells

目的：为揭示Ｃ５ａ和Ｃ５ａＲ对肾小球内皮细胞（ＧＶＥＣ）中性粒细胞（ＰＭＮ）粘附的影响及其影响程度。方法：采用免疫组化胶体金银染色检测Ｃ５ａＲ在人ＧＶＥＣ上的表达，并用微管吸吮技术定量测定了经Ｃ５ａ诱导后，ＧＶＥＣ和ＰＭＮ细胞对间的粘附力，同时通过测定粘附的ＰＭＮ中髓过氧化物酶（ＭＰＯ）含量变化，观察不同剂量Ｃ５ａ对粘附的影响。结果：发现ＧＶＥＣ上分布Ｃ５ａＲ，经２０００ｎｇ／ｍｌＣ５ａ刺激，ＧＶＥＣＰＭＮ间粘附力（Ｆａ）达００１４ｄｙｎ，相对粘附应力（Ｓ１）为１７２００ｄｙｎ／ｃｍ２，与对照组Ｆａ（０００１ｄｙｎ）和Ｓ１（３３９９ｄｙｎ／ｃｍ２）均有非常显著差异（Ｐ＜００１）。ＭＰＯ测定结果表明ＧＶＥＣＰＭＮ粘附的发生依赖于Ｃ５ａ剂量。结论：从生物力学角度直接定量阐明了Ｃ５ａＣ５ａＲ对ＧＶＥＣＰＭＮ粘附行为的重要影响。

Objective:To reveal whether C5a and C5aR affect the adhesion between human glomerulus vascular endothelial cells(GVEC) and granulocytes for determining.Methods:Immunogolden-sliver staining the expression of C5aR on GVEC,micropipette aspiration technique quantitatively studying the degree of the adhesion between GVEC and PMN and by measuring the change of amount of myeloperoxidase(MPO) in adherent PMN for analysing the dose of C5a which made the adhesion.Results:C5aR was expressed on human GVEC,when treating with 2 000 ng/ml C5a,the adhesive forces (Fa) and relative adhesion stress(S1) respectively reach 0 014 dyn and 1 720 0 dyn/cm 2,which had much markedly difference compared with that in controls(Fa=0.001 dyn,S1=339.9 dyn/cm 2).The adhesion between GVEC and PMN was enhanced in a C5a dose dependent manner.Conclusion:The data strongly directly suggested that C5a plays a key role in the adhesion between GVEC and PMN.