Clinton S K, Fleet J C, Loppnow H, Salomon R N, Clark B D, Cannon J G, Shaw A R, Dinarello C A, Libby P
Dana-Farber Cancer Institute, Boston, MA 02117.
Am J Pathol. 1991 Apr;138(4):1005-14.
Cultured human vascular endothelial and smooth muscle cells express interleukin-1 (IL-1) genes when exposed to bacterial lipopolysaccharides (LPS) and a variety of inflammatory mediators. Local production of IL-1 may contribute to the pathogenesis of various vascular diseases. Therefore the ability of intact vascular tissue to accumulate IL-1 mRNA and synthesize de novo biologically active IL-1 protein was examined. Escherichia coli LPS (10 micrograms/kg) was administered intravenously to adult rabbits and total RNA was isolated from aortic tissue at various times after LPS injection. In saline-injected rabbits, RNA extracted from the thoracic aorta contained little or no IL-1 message detected by Northern analysis using IL-1 alpha and beta cDNA probes cloned from an LPS-stimulated rabbit splenic macrophage library. Lipopolysaccharide treatment promptly induced transient accumulation of mRNA for IL-1 alpha and IL-1 beta within the aorta (maximal 1-hour after injection). Short-term organoid cultures of rabbit aorta exposed to LPS in vitro synthesized immunoprecipitable IL-1 alpha protein. Extracts of aortic tissue excised 1.5 to 3.0 hours after intravenous LPS administration contained immunoreactive and biologically active IL-1 alpha. Anti-rabbit IL-1 alpha antibody neutralized the biologic activity (more than 90%). Microscopic and immunohistochemical studies did not disclose adherent or infiltrating macrophages in rabbit aorta at the time of maximal IL-1 mRNA accumulation after LPS administration (1.5 hours), indicating that intrinsic vascular wall cells rather than mononuclear phagocytes probably account for the IL-1 activity induced by LPS. In addition, aortic tissue from rabbits fed an atherogenic diet showed an enhanced ability to accumulate IL-1 alpha and beta mRNA and produce immunodetectable protein in response to LPS administration. These studies demonstrate inducible IL-1 gene expression in rabbit vascular tissue in vivo and support a local role for this cytokine in vascular pathophysiology.
培养的人血管内皮细胞和平滑肌细胞在暴露于细菌脂多糖(LPS)和多种炎症介质时会表达白细胞介素-1(IL-1)基因。IL-1的局部产生可能促成各种血管疾病的发病机制。因此,研究了完整血管组织积累IL-1 mRNA并从头合成具有生物活性的IL-1蛋白的能力。将大肠杆菌LPS(10微克/千克)静脉注射给成年兔,并在LPS注射后的不同时间从主动脉组织中分离总RNA。在注射生理盐水的兔中,使用从LPS刺激的兔脾巨噬细胞文库克隆的IL-1α和β cDNA探针进行Northern分析,从胸主动脉提取的RNA几乎检测不到或没有检测到IL-1信息。脂多糖处理迅速诱导主动脉内IL-1α和IL-1β mRNA的瞬时积累(注射后1小时达到最大值)。体外暴露于LPS的兔主动脉短期类器官培养物合成了可免疫沉淀的IL-1α蛋白。静脉注射LPS后1.5至3.0小时切除的主动脉组织提取物含有免疫反应性和生物活性的IL-1α。抗兔IL-1α抗体中和了生物活性(超过90%)。显微镜和免疫组织化学研究未发现LPS给药后(1.5小时)IL-1 mRNA积累最大值时兔主动脉中有粘附或浸润的巨噬细胞,这表明内在的血管壁细胞而非单核吞噬细胞可能是LPS诱导的IL-1活性的原因。此外,喂食致动脉粥样硬化饮食的兔的主动脉组织显示,对LPS给药的反应中积累IL-1α和β mRNA以及产生可免疫检测蛋白的能力增强。这些研究证明了兔血管组织在体内可诱导的IL-1基因表达,并支持这种细胞因子在血管病理生理学中的局部作用。