Buchanan M R, Vazquez M J, Gimbrone M A
Blood. 1983 Oct;62(4):889-95.
Polymorphonuclear leukocytes (PMN) adhere to the vascular endothelial lining in vivo and to the surfaces of cultured endothelial cells in vitro, but the mechanisms of these cellular interactions remain unclear. Arachidonic acid metabolites, both cyclooxygenase- and lipoxygenase-derived, have been shown to influence PMN locomotion, secretion, and adhesion to artificial surfaces. To determine whether such mediators also are involved in regulating PMN-endothelial cell interactions, we have examined the effects of prostacyclin and various inhibitors of arachidonic acid metabolism on the adherence of radiolabeled PMN to cultured bovine aortic endothelial cells. Confluent endothelial monolayers were incubated with washed suspensions of radiolabeled human PMN (which contained less than 1% platelet contamination) at 37 degrees C for 30 min, then subjected to a standardized wash procedure and the number of adherent leukocytes determined radiometrically. Under basal conditions, i.e., in the absence of exogenous activating stimuli, 4,163 +/- 545 PMN adhered per square millimeter of endothelial surface (mean +/- SEM, n = 12). This basal adhesion (which corresponds to approximately 4-5 leukocytes per endothelial cell) was unaffected when the leukocytes and endothelial monolayers were pretreated with cyclooxygenase inhibitors (100 microM aspirin or 1-5 microM indomethacin) or PGI2 (10(-9)-10(6) M). Thus, basal PMN-endothelial adhesion in this in vitro model system does not appear to be dependent on endogenous cyclooxygenase derivatives of arachidonate or to be sensitive to inhibition by exogenous prostacyclin. In contrast, leukocyte adhesion was significantly reduced by pretreatment with 5,8,11,14- or 4,7,10,13-eicosatetraynoic acid, 0.5-5 mM sodium salicylate, or 10-1,000 microM indomethacin, antiinflammatory agents that can interfere with the metabolism of arachidonic acid via non-cyclooxygenase-dependent mechanisms. These observations may be relevant to the interactions of circulating PMN with vascular endothelium under both physiologic and pathophysiologic conditions in vivo.
多形核白细胞(PMN)在体内可黏附于血管内皮衬里,在体外可黏附于培养的内皮细胞表面,但其细胞间相互作用的机制仍不清楚。已证实,环氧化酶和脂氧化酶衍生的花生四烯酸代谢产物可影响PMN的运动、分泌及对人工表面的黏附。为确定此类介质是否也参与调节PMN与内皮细胞的相互作用,我们研究了前列环素及多种花生四烯酸代谢抑制剂对放射性标记的PMN黏附于培养的牛主动脉内皮细胞的影响。将汇合的内皮细胞单层与经洗涤的放射性标记人PMN悬浮液(血小板污染少于1%)在37℃孵育30分钟,然后进行标准化洗涤程序,并通过放射性测定法确定黏附白细胞的数量。在基础条件下,即在无外源性激活刺激时,每平方毫米内皮表面有4163±545个PMN黏附(平均值±标准误,n = 12)。当白细胞和内皮细胞单层用环氧化酶抑制剂(100μM阿司匹林或1 - 5μM吲哚美辛)或前列环素(10⁻⁹ - 10⁶M)预处理时,这种基础黏附(相当于每个内皮细胞约4 - 5个白细胞)不受影响。因此,在该体外模型系统中,基础PMN与内皮细胞的黏附似乎不依赖于花生四烯酸的内源性环氧化酶衍生物,对外源性前列环素的抑制也不敏感。相反,用5,8,11,14 - 或4,7,10,13 - 二十碳四烯酸、0.5 - 5 mM水杨酸钠或10 - 1000μM吲哚美辛预处理可显著降低白细胞黏附,这些抗炎药可通过非环氧化酶依赖机制干扰花生四烯酸的代谢。这些观察结果可能与体内生理和病理生理条件下循环PMN与血管内皮的相互作用有关。