用formononetin和salvianolic Acid B靶向RhoA表达，以减轻胰腺癌相关的内皮细胞变化

根据中药（TCM）中的气和血液理论，Qi强化草药和血液激活草药的结合在改善血液暂停综合征，尤其是在肿瘤治疗方面具有协同作用。经典的“ Radix Astragali -Salvia Miltiorrhiza”二人组典型地说明了这一原理，以振奋性气和激活血流而闻名，在肿瘤疗法中广泛使用。我们先前的研究强调了通过体外formononetin（来自Radix Astragali）和salvianolic Acid B（来自Salvia Miltiorrhiza）在体外结合使用胰腺肿瘤异种移植物的有效抑制作用。然而，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达到其抗癌作用。止血性血管系统，已知可促进肿瘤生长和转移。使肿瘤相关血管归一化的策略为抗肿瘤疗法提供了有希望的途径。这项研究旨在阐明formononetin与salvianolic Acid B（FC）结合在调节胰腺癌对内皮细胞的影响中的治疗潜力，阐明了控制这种治疗相互作用的潜在机制建立了（HUVEC）和PANC-1细胞，以研究将异常脉管系统靶向作为一种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）的单培养条件和共培养条件下的HUVEC增殖，迁移，侵袭和腔化发生。随后，评估了H-P系统的FCS处理其抗血管生成特性。分子对接被用来预测formononetin和salvianolic Acid b与RhoA之间的相互作用，并评估了HUVEC中RHOA的治疗后表达。此外，我们利用Shrhoa慢病毒来阐明RhoA在FCS介导的对HUVEC的影响中的作用。 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.分子对接分析表明FCS与RhoA之间的潜在相互作用。此外，发现FCS处理可下调RHOA的表达，并调节PI3K/AKT信号通路在PANC-1诱导的HUVEC中。值得注意的是，FCS对HUVEC的表型抑制作用被RhoA敲低减弱。体内斑马鱼研究验证了FCS的抗肿瘤活性，抑制了癌细胞的增殖，转移和血管发芽，同时促进了肿瘤细胞凋亡。这项研究强调了FC在胰腺癌引起的耐药性内皮层的痛苦中的有希望的潜力。 FCS表现出明显的抗抗血管效应，可以通过下调RhoA和抑制PI3K/AKT信号通路实现，从而提出了胰腺癌管理的新型治疗途径。

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.