Laboratory of Cardiovascular Biology, Department of Internal Medicine, University of Genova, Genova, Italy.
Oncology Unit, IRCCS AOU San Martino-IST, Genova, Italy.
Br J Pharmacol. 2017 Nov;174(21):3713-3726. doi: 10.1111/bph.13725. Epub 2017 Feb 22.
5-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.
EA.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 β-galactosidase and p16 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.
Both 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.
5FU 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.
This article is part of a themed section on New Insights into Cardiotoxicity Caused by Chemotherapeutic Agents. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.21/issuetoc.
5-氟尿嘧啶（5FU）及其前体药物卡培他滨可损伤血管内皮细胞，而胰高血糖素样肽 1（GLP-1）可维持内皮细胞完整性。本研究旨在探讨 5FU 对内皮细胞衰老的影响，以及 GLP-1 是否拮抗该作用。
EA.hy926 细胞分别用 5FU 或接受卡培他滨治疗的患者血清孵育，孵育前或孵育后用 GLP-1 预处理。衰老通过衰老相关的β-半乳糖苷酶和 p16 表达以及细胞增殖减少来鉴定。在接受卡培他滨化疗前后通过 ELISA 法测定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 CD146（内皮损伤标志物）。通过 RT-PCR、Western 印迹、信号转导抑制剂的功能实验和 ERK1/2 沉默，研究 5FU 诱导的表型并阐明 5FU 和 GLP-1 活性的作用途径。
5FU 和接受卡培他滨治疗的患者的血清均刺激内皮细胞衰老。5FU 诱导的衰老通过激活 p38 和 JNK 发生，与 eNOS 和 SIRT-1 水平降低有关。此外，5FU 上调 VCAM1 和 TYMP（编码激活卡培他滨和 5FU 的酶），且 sVCAM-1 和 CD146 浓度在卡培他滨化疗后高于化疗前。ICAM1 水平也有升高的趋势，但无统计学意义。GLP-1 拮抗 5FU 引发的衰老并降低 eNOS 和 SIRT-1 表达，这种保护作用由 GLP-1 受体、ERK1/2 介导，可能还由 PKA 和 PI3K 介导。
5FU 导致内皮细胞衰老和功能障碍，这可能导致其心血管副作用。GLP-1 可预防 5FU 诱导的衰老，提示使用 GLP-1 类似物和降解抑制剂治疗 5FU 和卡培他滨的血管毒性。
本文是关于“化疗药物引起的心脏毒性的新见解”专题的一部分。要查看该专题中的其他文章，请访问：http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.21/issuetoc.
