Xu Xiaoming, Wu Li, Zhou Xinbin, Zhou Nanyang, Zhuang Qin, Yang Jinxiu, Dai Jin, Wang Haibing, Chen Shenjie, Mao Wei
Department of Cardiology, Zhejiang Traditional Chinese Medical Hospital, Hangzhou, Zhejiang, China.
ShanXi Cardiovascular Hospital, Taiyuan, ShanXi, China.
Microvasc Res. 2017 May;111:25-31. doi: 10.1016/j.mvr.2016.12.011. Epub 2016 Dec 28.
Anti-angiogenesis has been proposed as an important strategy for angiogenesis-related diseases. Cryptotanshinone (CPT), an active component of Salvia miltiorrhiza, may be a potential inhibitor of angiogenesis. However, the molecular mechanisms underlying its anti-angiogenic activities remain poorly understood. This study is to investigate the effects of CPT on VEGF-induced angiogenesis and VEGFR2 signaling pathway in human umbilical vein endothelial cells (HUVECs).
HUVECs were treated with different concentration of CPT (5-20μmol/L) and the viability, endothelial cell migration, invasion, and tubular-like structure formation of HUVECs were detected by MTT, wound-healing migration, Transwell invasion and Matrigel tube formation assays, respectively. To assess the effect of CPT on VEGFR2 signaling pathway, VEGF-induced phosphorylation of VEGFR2 and its downstream molecules, including ERK1/2, p90RSK, Src and FAK were analyzed by Western blot.
CPT significantly suppressed VEGF-induced cells proliferation, migration, invasion, and tubular-like structure formation in HUVECs in a dose- and time-dependent manner. Western blot results revealed that CPT significantly suppressed VEGF-induced phosphorylation of VEGFR2 and its key downstream protein kinases, including p-ERK1/2, p-p90RSK, pY-Src and pY-FAK, which are responsible for endothelial cell migration, proliferation, and survival.
Our study suggested that CPT potently inhibits VEGF-induced angiogenesis by suppressing VEGFR2 activation and its downstream Src/FAK and ERK1/2 signaling pathways in HUVECs, highlighting the therapeutic potential for the treatment of angiogenesis-related diseases.
抗血管生成已被提出作为治疗血管生成相关疾病的重要策略。隐丹参酮（CPT）是丹参的一种活性成分，可能是一种潜在的血管生成抑制剂。然而，其抗血管生成活性的分子机制仍知之甚少。本研究旨在探讨CPT对人脐静脉内皮细胞（HUVECs）中VEGF诱导的血管生成及VEGFR2信号通路的影响。
用不同浓度（5 - 20μmol/L）的CPT处理HUVECs，分别通过MTT法、伤口愈合迁移实验、Transwell侵袭实验和基质胶管形成实验检测HUVECs的活力、内皮细胞迁移、侵袭及管状结构形成情况。为评估CPT对VEGFR2信号通路的影响，通过蛋白质免疫印迹法分析VEGF诱导的VEGFR2及其下游分子（包括ERK1/2、p90RSK、Src和FAK）的磷酸化情况。
CPT以剂量和时间依赖性方式显著抑制VEGF诱导的HUVECs细胞增殖、迁移、侵袭及管状结构形成。蛋白质免疫印迹结果显示，CPT显著抑制VEGF诱导的VEGFR2及其关键下游蛋白激酶（包括p-ERK1/2、p-p90RSK、pY-Src和pY-FAK）的磷酸化，这些蛋白激酶负责内皮细胞的迁移、增殖和存活。
我们的研究表明，CPT通过抑制HUVECs中VEGFR2激活及其下游Src/FAK和ERK1/2信号通路，有效抑制VEGF诱导的血管生成，凸显了其在治疗血管生成相关疾病方面的治疗潜力。
