根据中医气血理论，补气药与活血药配伍，对于改善血瘀证，尤其是治疗肿瘤有协同作用。经典的“黄芪丹参组合”就体现了这一理气活血的原理，广泛应用于肿瘤治疗。我们之前的研究强调了芒柄花素（来自黄芪）和丹酚酸 B（来自丹参）的组合在体外对胰腺肿瘤异种移植物的有效抑制作用。然而，目前尚不清楚这种组合是否可以抑制胰腺肿瘤的异常血管化以实现其抗癌作用。异常的血管化，已知会促进肿瘤的生长和转移。使肿瘤相关血管正常化的策略为抗肿瘤治疗提供了一条有希望的途径。本研究旨在揭示芒柄花素联合丹酚酸 B (FcS) 在调节胰腺癌对内皮细胞的影响方面的治疗潜力，阐明控制这种治疗相互作用的潜在机制，从而推进肿瘤血管系统正常化和对抗癌症进展的策略。建立了人脐静脉内皮细胞 (HUVEC) 和 PANC-1 细胞的共培养系统，以研究针对异常脉管系统作为新型抗肿瘤治疗策略的潜力。我们系统地比较了与 PANC-1 (H-P) 单独培养和共培养条件下的 HUVEC 增殖、迁移、侵袭和管腔生成。随后，评估了 H-P 系统的 FcS 处理的抗血管生成特性。利用分子对接预测芒柄花素和丹酚酸 B 与 RhoA 之间的相互作用，并评估 HUVEC 中 RhoA 治疗后的表达。此外，我们利用 shRhoA 慢病毒来阐明 RhoA 在 FcS 介导的 HUVEC 作用中的作用。在体内，采用斑马鱼异种移植肿瘤模型来评估 FcS 的抗肿瘤潜力，重点关注癌细胞增殖、迁移、凋亡和血管发育。FcS 治疗显示出对 PANC-1 诱导的肿瘤细胞改变的显着、剂量依赖性抑制。 HUVEC，包括增殖、迁移、侵袭和管形成能力。分子对接分析表明 FcS 和 RhoA 之间存在潜在的相互作用。此外，发现 FcS 处理可下调 PANC-1 诱导的 HUVEC 中的 RhoA 表达并调节 PI3K/AKT 信号通路。值得注意的是，FcS 对 HUVEC 的表型抑制作用因 RhoA 敲低而减弱。斑马鱼体内研究验证了 FcS 的抗肿瘤活性，抑制癌细胞增殖、转移和血管萌芽，同时促进肿瘤细胞凋亡。这项研究强调了 FcS 在对抗胰腺癌诱导的内皮细胞改变方面的巨大潜力。 FcS 表现出显着的抗异常脉管系统作用，可能通过下调 RhoA 和抑制 PI3K/Akt 信号通路来实现，从而为胰腺癌治疗提供了一种新的治疗途径。版权所有 © 2024。由 Elsevier B.V. 出版。

According to the theory of Qi and blood in Traditional Chinese Medicine (TCM), the combination of Qi-reinforcing herbs and blood-activating herbs has a synergistic effect in improving blood stasis syndrome, especially in tumor treatment. The classic "Radix Astragali - Salvia miltiorrhiza" duo exemplifies this principle, renowned for invigorating Qi and activating blood flow, employed widely in tumor therapies. Our prior research underscores the potent inhibition of pancreatic tumor xenografts by the combination of Formononetin (from Radix Astragali) and Salvianolic acid B (from Salvia miltiorrhiza) in vitro. However, it remains unclear whether this combination can inhibit the abnormal vascularization of pancreatic tumors to achieve its anti-cancer effect.Abnormal vasculature, known to facilitate tumor growth and metastasis. Strategies to normalize tumor-associated blood vessels provide a promising avenue for anti-tumor therapy. This study aimed to unravel the therapeutic potential of Formononetin combined with Salvianolic acid B (FcS) in modulating pancreatic cancer's impact on endothelial cells, illuminate the underlying mechanisms that govern this therapeutic interaction, thereby advancing strategies to normalize tumor vasculature and combat cancer progression.A co-culture system involving Human Umbilical Vein Endothelial Cells (HUVECs) and PANC-1 cells was established to investigate the potential of targeting abnormal vasculature as a novel anti-tumor therapeutic strategy. We systematically compared HUVEC proliferation, migration, invasion, and lumenogenesis in both mono- and co-culture conditions with PANC-1 (H-P). Subsequently, FcS treatment of the H-P system was evaluated for its anti-angiogenic properties. Molecular docking was utilized to predict the interactions between Formononetin and Salvianolic acid B with RhoA, and the post-treatment expression of RhoA in HUVECs was assessed. Furthermore, we utilized shRhoA lentivirus to elucidate the role of RhoA in FcS-mediated effects on HUVECs. In vivo, a zebrafish xenograft tumor model was employed to assess FcS's anti-tumor potential, focusing on cancer cell proliferation, migration, apoptosis, and vascular development.FcS treatment demonstrated a significant, dose-dependent inhibition of PANC-1-induced alterations in HUVECs, including proliferation, migration, invasion, and tube formation capabilities. Molecular docking analyses indicated potential interactions between FcS and RhoA. Further, FcS treatment was found to downregulate RhoA expression and modulated the PI3K/AKT signaling pathway in PANC-1-induced HUVECs. Notably, the phenotypic inhibitory effects of FcS on HUVECs were attenuated by RhoA knockdown. In vivo zebrafish studies validated FcS's anti-tumor activity, inhibiting cancer cell proliferation, metastasis, and vascular sprouting, while promoting tumor cell apoptosis.This study underscores the promising potential of FcS in countering pancreatic cancer-induced endothelial alterations. FcS exhibits pronounced anti-abnormal vasculature effects, potentially achieved through downregulation of RhoA and inhibition of the PI3K/Akt signaling pathway, thereby presenting a novel therapeutic avenue for pancreatic cancer management.Copyright © 2024. Published by Elsevier B.V.