Division of Vascular Surgery and Endovascular Services, SUNY Upstate Medical University, Syracuse, New York.
Department of Veterans Affairs, VA Healthcare Network Upstate, Syracuse, New York.
J Cell Biochem. 2020 Oct;121(10):4154-4165. doi: 10.1002/jcb.29686. Epub 2020 Apr 29.
The thrombospondins (TSPs), multifunctional matricellular proteins, are known mediators of endothelial cell (EC) angiogenesis and apoptosis. TSP-1, an antiangiogenic molecule, is important in the progression of vascular disease, in part by inducing EC apoptosis. TSP-2, although less studied, also induces EC apoptosis and inhibits angiogenesis. The effects of TSP-5 are largely unexplored in ECs, but TSP-5 is believed to be protective against arterial disease. Statin drugs have been shown to have beneficial pleiotropic effects, including decreasing EC apoptosis, increasing angiogenesis, and blocking TSP signaling. We hypothesized TSP-5 will be proangiogenic and antiapoptotic, and statin pretreatment would reverse the proapoptotic and antiangiogenic phenotype of TSP-1 and TSP-2. ECs were exposed to serum-free medium, TSP-1, TSP-2, or TSP-5 with or without fluvastatin pretreatment. Quantitative real-time polymerase chain reaction was performed on 96 apoptosis and 96 angiogenesis-related genes using microfluidic card assays. Angiogenesis was measured using Matrigel assays, while apoptosis was measured by fluorescent caspase assay. TSP-5 suppressed apoptotic genes and had a mixed effect on the angiogenic genes; however, TSP-5 did not alter apoptois but was proangiogenic. Pretreatment with fluvastatin downregulated proapoptotic genes and apoptosis and upregulated proangiogenic genes and angiogenesis. Findings indicate TSP-5 and fluvastatin have a protective effect on ECs, being proangiogenic and reversing the antiangiogenic effects of TSP-1 and TSP-2. In conclusion, TSP-5 and fluvastatin may be beneficial for inducing angiogenesis in the setting of ischemia.
血栓反应蛋白（TSPs）是多功能细胞外基质蛋白，已知其能介导内皮细胞（EC）血管生成和凋亡。TSP-1 是一种抗血管生成分子，在血管疾病的进展中很重要，部分原因是它诱导 EC 凋亡。TSP-2 虽然研究较少，但也诱导 EC 凋亡并抑制血管生成。TSP-5 在 EC 中的作用在很大程度上尚未得到探索，但 TSP-5 被认为可以预防动脉疾病。他汀类药物已被证明具有有益的多效性作用，包括减少 EC 凋亡、增加血管生成和阻断 TSP 信号。我们假设 TSP-5 将具有促血管生成和抗凋亡作用，他汀类药物预处理将逆转 TSP-1 和 TSP-2 的促凋亡和抗血管生成表型。将 EC 暴露于无血清培养基、TSP-1、TSP-2 或 TSP-5 中，同时或不预先用氟伐他汀处理。使用微流体卡测定法对 96 个凋亡和 96 个血管生成相关基因进行实时定量聚合酶链反应。通过 Matrigel 测定法测量血管生成，通过荧光半胱氨酸天冬氨酸测定法测量细胞凋亡。TSP-5 抑制凋亡基因，并对血管生成基因产生混合作用；然而，TSP-5 不会改变凋亡，但具有促血管生成作用。氟伐他汀预处理下调促凋亡基因和细胞凋亡，并上调促血管生成基因和血管生成。研究结果表明，TSP-5 和氟伐他汀对 EC 具有保护作用，具有促血管生成作用，并逆转 TSP-1 和 TSP-2 的抗血管生成作用。总之，TSP-5 和氟伐他汀可能有助于在缺血情况下诱导血管生成。
