5-fluorouracil causes endothelial cell senescence: potential protective role of glucagon-like peptide 1
5-fluorouracil causes endothelial cell senescence: potential protective role of glucagon-like peptide 1
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DOI：
10.1111/bph.13725
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发表时间：
2017-11-01
影响因子：
7.3
通讯作者：
Spallarossa, Paolo
中科院分区：
文献类型：
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作者：
Altieri, Paola;Murialdo, Roberto;Spallarossa, Paolo
背景与5-氟尿嘧啶(PURPOSE5-FU)及其前体药物卡培他滨可损伤内皮细胞，而胰高血糖素样肽1(GLP-1)可保护内皮细胞的完整性。在这里，我们研究了5FU对内皮细胞衰老的影响以及GLP-1是否可以拮抗它。实验用5FU或服用卡培他滨患者的血清暴露于Hy926细胞，并与GLP-1预先孵育或不预先孵育。衰老是通过衰老相关的β-半乳糖苷酶和p16(INK4a)的表达和细胞增殖减少来识别的。在卡培他滨化疗前和化疗结束时，用双抗体夹心酶联免疫吸附试验检测血清可溶性血管细胞黏附分子-1(sVCAM-1)、可溶性细胞间黏附分子-1(sICAM-1)和血管内皮细胞损伤标志物CD146。通过RT-PCR、Western blotting、信号转导抑制剂的功能实验和ERK1/2沉默来鉴定5FU诱导的表型，并阐明5FU和GLP-1活性的途径。5FU诱导的衰老通过激活p38和JNK而发生，并与eNOS和SIRT-1水平降低有关。此外，5FU上调VCAM1和TYMP(编码酶激活的卡培他滨和5FU)，sVCAM-1和CD146浓度高于卡培他滨化疗前。ICAM1水平升高的趋势也不明显。GLP-1拮抗5FU诱导的衰老，减少eNOS和SIRT-1的表达，这种保护作用可能通过GLP-1受体ERK1/2以及可能的PKA和PI3K介导。结论GLP-1和5FU可导致内皮细胞衰老和功能障碍，这可能是其心血管副作用的原因之一。GLP-1可阻止5FU引发的衰老，增加了使用GLP-1类似物和降解抑制剂治疗5FU和卡培他滨血管毒性的可能性。
BACKGROUND AND PURPOSE5-fluorouracil (5FU) and its prodrug, capecitabine, can damage endothelial cells, whilst endothelial integrity is preserved by glucagon-like peptide 1 (GLP-1). Here, we studied the effect of 5FU on endothelial senescence and whether GLP-1 antagonizes it.EXPERIMENTAL APPROACHEA.hy926 cells were exposed to 5FU or sera from patients taking capecitabine, with or without pre-incubation with GLP-1. Senescence was identified by expression of senescence-associated beta-galactosidase and p16(INK4a) and reduced cell proliferation. Soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1) and CD146 (marker of endothelial injury) were measured by ELISA before and at completion of capecitabine chemotherapy. RT-PCR, western blotting, functional experiments with signalling inhibitors and ERK1/2 silencing were performed to characterize 5FU-induced phenotype and elucidate the pathways underlying 5FU and GLP-1 activity.KEY RESULTSBoth 5FU and sera from capecitabine-treated patients stimulated endothelial cell senescence. 5FU-elicited senescence occurred via activation of p38 and JNK, and was associated with decreased eNOS and SIRT-1 levels. Furthermore, 5FU up-regulated VCAM1 and TYMP (encodes enzyme activating capecitabine and 5FU), and sVCAM-1 and CD146 concentrations were higher after than before capecitabine chemotherapy. A non-significant trend for higher ICAM1 levels was also observed. GLP-1 counteracted 5FU-initiated senescence and reduced eNOS and SIRT-1 expression, this protection being mediated by GLP-1 receptor, ERK1/2 and, possibly, PKA and PI3K.CONCLUSIONS AND IMPLICATIONS5FU causes endothelial cell senescence and dysfunction, which may contribute to its cardiovascular side effects. 5FU-triggered senescence was prevented by GLP-1, raising the possibility of using GLP-1 analogues and degradation inhibitors to treat 5FU and capecitabine vascular toxicity.