Warren J B, Brady A J, Taylor G W
Department of Clinical Pharmacology, Royal Postgraduate Medical School, London, U.K.
Proc Biol Sci. 1990 Aug 22;241(1301):127-31. doi: 10.1098/rspb.1990.0076.
Conditioned medium was collected from vascular smooth-muscle cells grown in culture to determine if these cells synthesize vasoactive substances. The medium caused a short-acting endothelium-independent constriction of rat aorta, followed by a prolonged, endothelium-dependent relaxation. This relaxation was mediated through the release of endothelium-derived relaxing factor (EDRF) as it was abolished by the addition of methylene blue (5 x 10(-6) M), haemoglobin (10(-6) M) or methyl arginine, but was not affected by indomethacin (10(-5) M). Smooth-muscle medium stimulated the production of EDRF from both rat and rabbit thoracic aortic rings as well as from cultured bovine pulmonary artery endothelial cells. The prolonged stimulation of EDRF by smooth-muscle medium was not mimicked by known physiological stimuli to EDRF release; EDRF-stimulating activity was not affected when smooth-muscle cells were grown in the presence of indomethacin (10(-5) M), although serum in the medium was required. The EDRF-stimulating substance(s) in the smooth-muscle medium was heat stable and associated with a high molecular mass (30,000 greater than Mr greater than 3500) water-soluble species that is as yet unidentified.
收集培养的血管平滑肌细胞的条件培养基，以确定这些细胞是否合成血管活性物质。该培养基引起大鼠主动脉短暂的非内皮依赖性收缩，随后是长时间的内皮依赖性舒张。这种舒张是通过内皮衍生舒张因子（EDRF）的释放介导的，因为添加亚甲蓝（5×10⁻⁶ M）、血红蛋白（10⁻⁶ M）或甲基精氨酸可消除这种舒张，但吲哚美辛（10⁻⁵ M）对此无影响。平滑肌培养基刺激大鼠和兔胸主动脉环以及培养的牛肺动脉内皮细胞产生EDRF。平滑肌培养基对EDRF的长时间刺激不能被已知的EDRF释放生理刺激所模拟；当平滑肌细胞在吲哚美辛（10⁻⁵ M）存在的情况下生长时，EDRF刺激活性不受影响，尽管培养基中的血清是必需的。平滑肌培养基中的EDRF刺激物质是热稳定的，并且与一种尚未鉴定的高分子量（Mr大于3500且大于30,000）水溶性物质相关。
