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利用５１Ｃｒ－标记血小板检测其与内皮细胞粘附功能；ＥＬＩＳＡ方法检测血管内皮细胞表面玻璃连接蛋白受体（ＶｎＲ）表达改变；Ｆｕｒａ－２／Ａｍ负载测定群体细胞胞浆游离钙；Ｆｌｕｏ－３／Ａｍ负载测定单细胞内游离钙等方法，观察了肿瘤坏死因子（ＴＮＦ）对内皮细胞（ＥＣ）与血小板粘附功能的影响，发现ＴＮＦ－α（１００Ｕ／ｍｌ）可明显促进血小板（ＰＬ）与ＥＣ的粘附，ｃｐｍ值明显高于对照组（４１０．７±１７．６ｖｓ．２１９．７±１６．３ｎ＝６Ｐ＜０．０１）．用抗ＶｎＲ－β３亚单位的单抗（ＭｃＡｂ）可大部分阻断ＴＮＦ－α的促粘附作用，与未加单抗的ＴＮＦα组相比有显著差异，非ＭｃＡｂ组ｃｐｍ为４１０．５±１７．６，ＭｃＡｂ组为２８８．３±１３．８，（ｎ＝６，Ｐ＜０．０１），用ＥＬＩＳＡ法证实不同浓度ＴＮＦ－α（２００Ｕ／ｍｌ～１０００Ｕ／ｍｌ）可增加内皮细胞表面ＶｎＲ表达，以１０００Ｕ／ｍｌ组为最明显。同一浓度ＴＮＦ－α组，ＶｎＲ的表达呈现随作用时间延长而增加的趋势。此外，ＴＮＦ－α可增加ＥＣ［Ｃａ２＋］ｉ，群体细胞和单个细胞测定的结果一致。以上结果提示，ＴＮＦ－α可增加ＥＣ与血小板的粘附功能，其作用机制是以ＶｎＲ－（αｖβ３?
The adhesion of endothelial cells was detected by 51Cr-labeled platelets, the expression of vitronectin receptor (VnR) was detected by ELISA, the cytoplasmic free calcium was detected by Fura-2 / Am loading, the expression of Fluo-3 / Am load determination of intracellular free calcium and other methods to observe the effect of tumor necrosis factor (TNF) on endothelial cells (EC) and platelet adhesion function and found that TNF-α (100U / ml) can significantly promote platelet EC adhesion, cpm values were significantly higher than the control group (410.7 ± 17.6vs.219.7 ± 16.3n = 6P <0.01). The anti-VnR-β3 subunit monoclonal antibody (McAb) can block the adhesion of TNF-α, which is significantly different from the non-McAb TNFα group. The non-McAb cpm was 410.5 ± 17.6 in the McAb group and 288.3 ± 13.8 in the McAb group (n = 6, P <0.01). It was confirmed by ELISA that different concentrations of TNF-α (200U / ml ~ 1000U / ml) VnR expression, with 1000U / ml group was the most obvious. The same concentration of TNF-α group, VnR expression showed an increasing trend with time. In addition, TNF-α increased EC [Ca2 +] i, consistent with the results of population cell and single cell assays. The above results suggest that TNF-α can increase the EC and platelet adhesion function, its mechanism of action is VnR- (αvβ3?