Sánchez-Crespo M, Iñarrea P, Alvarez V, Alonso F, Egido J, Hernando L
Am J Physiol. 1983 Jun;244(6):F706-11. doi: 10.1152/ajprenal.1983.244.6.F706.
Urine from normotensive volunteers and patients with systemic lupus erythematosus glomerulonephropathy was sequentially concentrated by negative-pressure ultrafiltration, dialyzed against distilled water, and extracted into the chloroform phase of a mixture of organic solvents(chloroform:methanol:water, 1:1:0.9 vol/vol). The lipid fraction was further purified by thin-layer chromatography on silica gel plates using neutral, acidic, and basic mixtures of organic solvents and it was then tested for its ability to induce the release of [3H]serotonin from rabbit platelets. All of the samples contained a platelet-activating moiety similar to a synthetic platelet-activating factor (PAF-acether) on the basis of its chromatographic behavior, resistance to the pretreatment of platelets by 10(-6) M indomethacin, and loss of activity by alkaline methanolysis or treatment by phospholipases A2, C, and D. Cross-densensitization experiments between synthetic PAF-acether and the urine factor showed that both compounds act on platelets through the activation of the same putative receptor. Further, the urine factor induced hypotension when intra-arterially injected in normotensive rats, and this activity was also abrogated by alkaline methanolysis. In summary, these data provide evidence of the presence in normal human urine and, probably, of the release by the kidney of a lipid factor with platelet-activating and hypotensive activity whose general structure seems to be alkyl-acyl-glyceryl-phosphorylcholine and, therefore, is similar to the structure of the inflammatory mediator PAF-acether and the antihypertensive polar renomedullary lipid.
来自血压正常志愿者和系统性红斑狼疮性肾小球肾炎患者的尿液,首先通过负压超滤进行浓缩,然后用蒸馏水透析,再萃取到有机溶剂混合物(氯仿:甲醇:水,1:1:0.9体积/体积)的氯仿相中。脂质部分通过在硅胶板上进行薄层层析进一步纯化,使用中性、酸性和碱性有机溶剂混合物,然后测试其诱导兔血小板释放[3H]5-羟色胺的能力。基于其色谱行为、对10(-6) M消炎痛预处理血小板的抗性以及碱性甲醇解或磷脂酶A2、C和D处理导致的活性丧失,所有样品均含有一种类似于合成血小板激活因子(PAF-乙醚)的血小板激活部分。合成PAF-乙醚与尿液因子之间的交叉脱敏实验表明,这两种化合物都通过激活相同的假定受体作用于血小板。此外,尿液因子经动脉内注射到血压正常的大鼠体内时会引起低血压,这种活性也会因碱性甲醇解而消除。总之,这些数据证明了正常人类尿液中存在一种脂质因子,并且可能是由肾脏释放的,该脂质因子具有血小板激活和降压活性,其总体结构似乎是烷基-酰基-甘油-磷酸胆碱,因此与炎症介质PAF-乙醚和抗高血压极性肾髓质脂质的结构相似。