Lipoproteins containing apoprotein B are a major regulator of neutrophil responses to monosodium urate crystals.
Lipoproteins containing apoprotein B are a major regulator of neutrophil responses to monosodium urate crystals.
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含有脱辅基蛋白 B 的脂蛋白是中性粒细胞对尿酸钠晶体反应的主要调节因子。
DOI：
10.1172/jci111380
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发表时间：
1984
期刊：
影响因子：
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通讯作者：
Ginsberg,MH
中科院分区：
文献类型：
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作者：
Terkeltaub,R;Curtiss,LK;Tenner,AJ;Ginsberg,MH
痛风性关节炎患者对关节内尿酸盐结晶的炎症反应是不同的。可变性的一个来源可能是晶体结合蛋白对细胞反应的调节。我们已经在与血浆中的尿酸盐晶体结合的多肽中鉴定出三种载脂蛋白。鉴定首先基于它们与分离的脂蛋白中的多肽的共电泳性，以及暴露在脂蛋白耗尽血浆中的晶体的蛋白外壳的减少。通过免疫印迹技术将这些载脂蛋白鉴定为载脂蛋白A-I、载脂蛋白B(Apo B)和载脂蛋白E。由于中性粒细胞在急性痛风中起核心作用，我们研究了脂蛋白对中性粒细胞-尿酸盐晶体相互作用的影响。血浆可显著抑制尿酸盐晶体诱导的中性粒细胞鲁米诺依赖的化学发光(CL)。KbR密度梯度离心法去除脂蛋白可完全消除血浆对尿酸盐诱导的CL的抑制作用。通过重新添加小于或等于1.25g/cm~3脂蛋白分数的d，可恢复脂蛋白耗竭血浆的抑制活性。血浆还抑制尿酸盐晶体诱导的中性粒细胞超氧化物生成和细胞溶解(乳酸脱氢酶丢失)。这种抑制作用被脂蛋白耗竭显著减弱，这表明脂蛋白的作用并不局限于CL。用极低、中、低密度脂蛋白(LDL)重组的去脂蛋白血浆可抑制晶体诱导的CL。高密度脂蛋白重建无效。琼脂糖基结合的载脂蛋白B特异性抗体去除血浆中所有载脂蛋白，也消除了对尿酸盐诱导的CL的所有抑制活性。因此，载脂蛋白B被证明是血浆中的抑制物种。在没有其他血浆蛋白的情况下，载脂蛋白B与尿酸盐晶体的结合和CL的抑制也被观察到。此外，除了载脂蛋白外，还通过检测与晶体相关的胆固醇来验证整个脂蛋白颗粒与晶体的结合。低密度脂蛋白对尿酸盐晶体诱导的CL的作用是刺激特异性的。尿酸盐晶体与中性粒细胞与10微克/毫升低密度脂蛋白共同孵育后，抑制率达83%。相反，低密度脂蛋白不抑制化学趋化性六肽、调理酵母多糖和金黄色葡萄球菌的CL反应。载脂蛋白B耗竭对尿酸盐晶体诱导的CL的血浆抑制作用似乎是独一无二的。去除其他尿酸盐晶体结合蛋白的血浆或血清，包括纤维蛋白原、纤维连接蛋白、C1q和免疫球蛋白，几乎保留了它们所有的CL抑制活性。因此，含有载脂蛋白B的脂蛋白是中性粒细胞对尿酸盐晶体反应的主要调节因子。这些脂蛋白以不同浓度存在于滑液中，并可能对痛风的病程产生重要影响。
The inflammatory response to intraarticular urate crystals is known to be variable in gouty arthritis. One source of variability may be the modulation of cellular responses by crystal-bound proteins. We have identified three apolipoproteins among the polypeptides bound to urate crystals exposed to plasma. Identification was first based on their coelectrophoresis with polypeptides from isolated lipoproteins and diminution in the protein coat of crystals exposed to lipoprotein-depleted plasma. The apoproteins were immunochemically identified by the Western blotting technique as apoprotein A-I, apoprotein B (apo B), and apoprotein E. Because neutrophils play a central role in acute gout, we investigated the potential effects of lipoproteins on neutrophil-urate crystal interactions. Plasma profoundly inhibited urate crystal-induced neutrophil luminol-dependent chemiluminescence (CL). Lipoprotein depletion by KBr density gradient centrifugation completely abrogated the inhibitory effect of plasma on urate-induced CL. The inhibitory activity of lipoprotein-depleted plasma was restored by adding back the d less than or equal to 1.25 g/cm3 lipoprotein fraction. Plasma also inhibited urate crystal-induced neutrophil superoxide generation and cytolysis (lactic dehydrogenase loss). This inhibition was significantly diminished by lipoprotein depletion, indicating that the lipoprotein effect was not limited to CL. Lipoprotein-depleted plasma reconstituted with very low, intermediate, and low density lipoproteins (LDL) inhibited crystal-induced CL. High density lipoprotein reconstitution was without effect. Immunodepletion from plasma of all apo B lipoproteins by agarose-bound apo B-specific antibody also removed all inhibitory activity for urate-induced CL. Thus, apo B lipoproteins were shown to be the inhibitory species in plasma. Binding of apo B lipoproteins to urate crystals and inhibition of CL was also seen in the absence of other plasma proteins. In addition, the binding of whole lipoprotein particles to the crystals was verified by detection of crystal-associated cholesterol in addition to the apoprotein. The effects of LDL on urate crystal-induced CL were stimulus specific. Coincubation of urate crystals and neutrophils in the presence of 10 micrograms/ml LDL resulted in 83% inhibition. In contrast, CL responses to a chemotactic hexapeptide, opsonized zymosan, and Staphylococcus aureus were not inhibited by LDL. The effects of depletion of apo B lipoproteins on plasma suppression of urate crystal-induced CL appeared to be unique. Plasma or sera depleted of other urate crystal-binding proteins including fibrinogen, fibronectin, C1q, and IgG retained virtually all their CL inhibitory activity. Lipoproteins containing apo B are thus a major regulator of neutrophil responses to urate crystals. These lipoproteins are present in variable concentration in synovial fluid and may exert an important influence on the course of gout.Images