Arriero Maria M, de La Pinta Juan C, Escribano Marta, Celdrán Angel, Muñoz-Alameda Luis, García-Cañete Joaquin, Jiménez Ana M, Casado Santos, Farré Jerónimo, López-Farré Antonio
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Circ Res. 2002 Apr 5;90(6):719-27. doi: 10.1161/01.res.0000013699.74563.42.
The aim was to analyze whether pericardial tissue expresses endothelial NO synthase (eNOS) protein and to determine the presence of cytosolic proteins that bind to eNOS mRNA. The effect of aspirin on the above-mentioned parameters was also analyzed. eNOS protein was expressed in pericardial tissue from male guinea pigs. Escherichia coli lipopolysaccharide (LPS, 10 microgram/mL) and Staphylococcus aureus endotoxin (SA, 10 microgram/mL) reduced eNOS protein expression and shortened the half-life of the eNOS messenger. Under basal conditions, cytosolic extracts from pericardial samples bound to the 3'-untranslated region (3'-UTR) of eNOS mRNA, which was enhanced by LPS and SA. Proteinase K fully prevented the binding of cytosolic pericardial extracts to 3'-UTR of eNOS mRNA, suggesting the involvement of proteins that were further characterized as 60- and 51-kDa proteins. Aspirin (1 to 10 mmol/L) restored eNOS expression in either LPS- and SA-stimulated pericardial samples and reduced the binding activity of the pericardial cytosolic proteins to 3'-UTR of eNOS mRNA. Indomethacin also reduced the downregulation of eNOS by LPS and diminished the binding activity of the cytosolic proteins, although higher doses of indomethacin than of aspirin were needed to improve these parameters. In conclusion, eNOS protein is expressed in guinea pig pericardial tissue. LPS and SA stimulate the binding activity of pericardial cytosolic proteins to 3'-UTR of eNOS mRNA and reduce eNOS protein expression. High doses of aspirin and indomethacin protect eNOS protein expression and reduce the binding activity of the cytosolic proteins to 3'-UTR of eNOS mRNA, suggesting an inverse association between the presence of these cytosolic proteins and eNOS expression.
目的是分析心包组织是否表达内皮型一氧化氮合酶（eNOS）蛋白，并确定与eNOS mRNA结合的胞质蛋白的存在情况。同时还分析了阿司匹林对上述参数的影响。雄性豚鼠的心包组织中表达了eNOS蛋白。大肠杆菌脂多糖（LPS，10微克/毫升）和金黄色葡萄球菌内毒素（SA，10微克/毫升）可降低eNOS蛋白表达，并缩短eNOS信使的半衰期。在基础条件下，心包样本的胞质提取物与eNOS mRNA的3'-非翻译区（3'-UTR）结合，LPS和SA可增强这种结合。蛋白酶K完全阻止了心包胞质提取物与eNOS mRNA的3'-UTR结合，表明存在进一步鉴定为60 kDa和51 kDa的蛋白质参与其中。阿司匹林（1至10毫摩尔/升）可恢复LPS和SA刺激的心包样本中的eNOS表达，并降低心包胞质蛋白与eNOS mRNA的3'-UTR的结合活性。吲哚美辛也可减少LPS对eNOS的下调作用，并降低胞质蛋白的结合活性，尽管需要比阿司匹林更高剂量的吲哚美辛才能改善这些参数。总之，豚鼠心包组织中表达eNOS蛋白。LPS和SA刺激心包胞质蛋白与eNOS mRNA 的3'-UTR的结合活性，并降低eNOS蛋白表达。高剂量的阿司匹林和吲哚美辛可保护eNOS蛋白表达，并降低胞质蛋白与eNOS mRNA的3'-UTR的结合活性，表明这些胞质蛋白的存在与eNOS表达之间存在负相关。
