Department of Experimental and Clinical Pharmacology, College of Pharmacy, University of Minnesota, Minneapolis, MN 55455, USA.
Cells. 2022 Jun 21;11(13):1992. doi: 10.3390/cells11131992.
Doxorubicin (DOX) induces endothelial cell (EC) senescence, which contributes to endothelial dysfunction and cardiovascular complications. Senolytic drugs selectively eliminate senescent cells to ameliorate senescence-mediated pathologies. Previous studies have demonstrated differences between immortalized and primary EC models in some characteristics. However, the response of DOX-induced senescent ECs to senolytics has not been determined across these two models. In the present work, we first established a comparative characterization of DOX-induced senescence phenotypes in immortalized EA.hy926 endothelial-derived cells and primary human umbilical vein EC (HUVECs). Thereafter, we evaluated the senolytic activity of four senolytics across both ECs. Following the DOX treatment, both EA.hy926 and HUVECs shared similar senescence phenotypes characterized by upregulated senescence markers, increased SA-β-gal activity, cell cycle arrest, and elevated expression of the senescence-associated secretory phenotype (SASP). The potentially senolytic drugs dasatinib, quercetin, and fisetin demonstrated a lack of selectivity against DOX-induced senescent EA.hy926 cells and HUVECs. However, ABT-263 (Navitoclax) selectively induced the apoptosis of DOX-induced senescent HUVECs but not EA.hy926 cells. Mechanistically, DOX-treated EA.hy926 cells and HUVECs demonstrated differential expression levels of the BCL-2 family proteins. In conclusion, both EA.hy926 cells and HUVECs demonstrate similar DOX-induced senescence phenotypes but they respond differently to ABT-263, presumably due to the different expression levels of BCL-2 family proteins.
多柔比星（DOX）诱导内皮细胞（EC）衰老，这有助于内皮功能障碍和心血管并发症。衰老细胞选择性消除药物（Senolytics）可选择性地消除衰老细胞，从而改善衰老相关的病变。先前的研究已经证明在某些特性上，永生化和原代 EC 模型之间存在差异。然而，两种模型中 DOX 诱导的衰老 EC 对 Senolytics 的反应尚未确定。在本研究中，我们首先比较了永生化 EA.hy926 内皮细胞和原代人脐静脉 EC（HUVEC）中 DOX 诱导的衰老表型。此后，我们评估了四种 Senolytics 在两种 EC 中的 Senolytic 活性。在 DOX 处理后，EA.hy926 和 HUVEC 均表现出相似的衰老表型，其特征是衰老标志物上调、SA-β-gal 活性增加、细胞周期停滞和衰老相关分泌表型（SASP）的表达升高。潜在的 Senolytic 药物达沙替尼、槲皮素和非瑟酮对 DOX 诱导的 EA.hy926 细胞和 HUVEC 衰老缺乏选择性。然而，ABT-263（Navitoclax）选择性诱导 DOX 诱导的衰老 HUVEC 凋亡，但不诱导 EA.hy926 细胞凋亡。在机制上，用 DOX 处理的 EA.hy926 细胞和 HUVEC 显示出 BCL-2 家族蛋白的表达水平不同。总之，EA.hy926 细胞和 HUVEC 均表现出相似的 DOX 诱导的衰老表型，但对 ABT-263 的反应不同，这可能是由于 BCL-2 家族蛋白的表达水平不同。
