Liagre B, Vergne P, Rigaud M, Beneytout J L
Faculté de Médecine, ERS CNRS 6101, Limoges, France.
FEBS Lett. 1997 Sep 1;414(1):159-64. doi: 10.1016/s0014-5793(97)00904-6.
In the present study, we have demonstrated platelet-type 12-lipoxygenase (12-LOX) expression in human rheumatoid arthritis (RA) type B synoviocytes by reverse-transcription polymerase chain reaction (RT-PCR). The presence of 12-LOX mRNA in these cells was revealed by classical RT-PCR analysis using platelet-type 12-LOX cDNA primers and the PCR fragment (246 bp) was purified, amplified and sequenced. By sequence analysis, this fragment was determined to be 100% identical to that from platelet-type 12-LOX cDNA. Immunofluorescence data demonstrate that interleukin-1beta (IL-1beta) increases cellular 12-LOX protein. Other results associate specific inflammatory cytokines with the activity of 12-LOX in human RA type B synoviocytes. IL-1beta increased 12S-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) production (4-fold) and we also observed an increase in 12-HETE production (2.5-fold) after incubation of human RA type B synoviocytes with TNF alpha. In contrast to the action of IL-1beta on 12-HETE synthesis, IL-4 and IL-6 did not enhance 12-HETE production. This is the first demonstration of platelet-type 12-LOX cDNA derived from the mRNA of cultured human RA type B synoviocytes.
在本研究中，我们通过逆转录聚合酶链反应（RT-PCR）证明了血小板型12-脂氧合酶（12-LOX）在人类风湿性关节炎（RA）B型滑膜细胞中的表达。使用血小板型12-LOX cDNA引物通过经典RT-PCR分析揭示了这些细胞中12-LOX mRNA的存在，并且纯化、扩增和测序了PCR片段（246 bp）。通过序列分析，确定该片段与血小板型12-LOX cDNA的片段100%相同。免疫荧光数据表明，白细胞介素-1β（IL-1β）可增加细胞12-LOX蛋白。其他结果表明，特定的炎性细胞因子与人RA B型滑膜细胞中12-LOX的活性相关。IL-1β使12S-羟基-5,8,10,14-二十碳四烯酸（12-HETE）的产生增加（4倍），并且在用人RA B型滑膜细胞与肿瘤坏死因子α（TNFα）孵育后，我们还观察到12-HETE的产生增加（2.5倍）。与IL-1β对12-HETE合成的作用相反，IL-4和IL-6并未增强12-HETE的产生。这是首次证明从培养的人RA B型滑膜细胞的mRNA衍生出血小板型12-LOX cDNA。
