Deacon C F, Johnsen A H, Holst J J
Department of Medical Physiology, Panum Institute, University of Copenhagen, Denmark.
J Clin Endocrinol Metab. 1995 Mar;80(3):952-7. doi: 10.1210/jcem.80.3.7883856.
The metabolism of glucagon-like peptide-1 (GLP-1) has not been studied in detail, but it is known to be rapidly cleared from the circulation. Measurement by RIA is hampered by the fact that most antisera are side-viewing or C-terminally directed, and recognize both intact GLP-1 and biologically inactive. N-terminally truncated fragments. Using high pressure liquid chromatography in combination with RIAs, methodology allowing specific determination of both intact GLP-1 and its metabolites was developed. Human plasma was shown to degrade GLP-1-(7-36)amide, forming an N-terminally truncated peptide with a t1/2 of 20.4 +/- 1.4 min at 37 C (n = 6). This was unaffected by EDTA or aprotinin. Inhibitors of dipeptidyl peptidase-IV or low temperature (4 C) completely prevented formation of the metabolite, which was confirmed to be GLP-1-(9-36)amide by mass spectrometry and sequence analysis. High pressure liquid chromatography revealed the concentration of GLP-1-(9-36)amide to be 53.5 +/- 13.7% of the concentration of endogenous intact GLP-1 in the fasted state, which increased to 130.8 +/- 10.0% (P < 0.01; n = 6) 1 h postprandially. Metabolism at the C-terminus was not observed. This study suggests that dipeptidyl peptidase-IV is the primary mechanism for GLP-1 degradation in human plasma in vitro and may have a role in inactivating the peptide in vivo.
胰高血糖素样肽-1(GLP-1)的代谢尚未得到详细研究,但已知它会迅速从循环中清除。放射免疫分析(RIA)测量受到阻碍,因为大多数抗血清是侧视或C末端定向的,并且能识别完整的GLP-1和无生物学活性的N末端截短片段。使用高压液相色谱结合放射免疫分析,开发了一种能够特异性测定完整GLP-1及其代谢产物的方法。研究表明,人血浆会降解GLP-1-(7-36)酰胺,在37℃下形成一种N末端截短的肽,其半衰期为20.4±1.4分钟(n = 6)。这不受乙二胺四乙酸(EDTA)或抑肽酶的影响。二肽基肽酶-IV抑制剂或低温(4℃)完全阻止了代谢产物的形成,通过质谱和序列分析证实该代谢产物为GLP-1-(9-36)酰胺。高压液相色谱显示,在禁食状态下,GLP-1-(9-36)酰胺浓度为内源性完整GLP-1浓度的53.5±13.7%,餐后1小时该比例增至130.8±10.0%(P < 0.01;n = 6)。未观察到C末端的代谢情况。该研究表明,二肽基肽酶-IV是体外人血浆中GLP-1降解的主要机制,可能在体内使该肽失活中发挥作用。