Danggui buxue decoction attenuates 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone - induced lung cancer growth in A/J mice by suppressing HIF-1α/VE GF-mediated angiogenesis

Wu Chen¹Ying Guo^1,2*Huan Liu³Shanyu Zhang⁴Sanyin Zhang²Zhilong Liu¹

• ¹Guangzhou University of Traditional Chinese Medicine First Affiliated Hospital Chongqing Hospital (Chongqing Beibei Traditional Chinese Medicine Hospital), chong qing, China
• ²Chengdu University of TCM, Chengdu, China
• ³Taian City Central Hospital, Tai'an, China
• ⁴Outpatient Department of the 38th Ex-Cadre Sanatorium, Beijing, China

Background: Lung cancer (LC) persists as a leading cause of global cancer-related mortality. Pathological angiogenesis constitutes a critical mechanism in LC progression, facilitating neovascularization that supplies oxygen and nutrients to support tumor growth. Despite this, current anti-angiogenic therapies face significant clinical limitations. Danggui Buxue Decoction (DBD), a traditional Chinese herbal formula used to tonify Qi and activate blood circulation, exhibits clinical potential in delaying LC progression; however, its precise mechanistic basis remains incompletely defined. This study aimed to evaluate the inhibitory effects of DBD aqueous extract on lung tumors in 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced A/J mice and elucidate whether its therapeutic mechanism involves suppression of hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor-A (VEGF)-mediated angiogenesis. Methods: Potential therapeutic targets of DBD against LC were identified through database mining (OMIM, TTD, GeneCards) using topological analysis. A/J mice received intraperitoneal injections of NNK (100 mg/kg) to induce lung tumors. From week 10, DBD aqueous extract (10 g/kg/day) was administered via oral gavage. Lung tumor progression and systemic parameters were assessed at weeks 10 and 20 using small-animal CT, enzyme-linked immunosorbent assay (ELISA), Doppler ultrasound, pulmonary function tests, and complete blood counts. At week 20, mice were anesthetized with 3% isoflurane and sacrificed by cervical dislocation. Lung tissues were harvested for histopathological evaluation (H&E), immunohistochemistry (CD31), and immunofluorescence (HIF-1α/VEGF) to quantify microvessel density and hypoxia/angiogenesis markers. Results: Network pharmacology identified TP53, AKT1, MYC, and VEGF as core therapeutic targets of DBD. In vivo, small-animal CT detected pulmonary opacities at week 10, concomitant with elevated pulmonary artery flow, increased airway resistance, and heightened circulating levels of TNF-α, VEGF, white blood cells (WBC), and neutrophils. By week 20, progressive multifocal opacities emerged alongside reduced pulmonary artery flow, impaired lung function, elevated TNF-α/VEGF, and decreased WBC, lymphocytes, and platelets. Compared to untreated controls, 10-week DBD treatment significantly suppressed lung tumor growth, reduced lesion microvessel density, downregulated HIF-1α and VEGF expression, and ameliorated hematological dysregulation. Conclusion: Our findings indicate that angiogenesis serves as a core mechanism driving NNK-induced lung tumorigenesis in mice. DBD attenuates tumor growth primarily by inhibiting HIF-1α/VEGF-mediated angiogenesis, with complementary contributions from restored immune homeostasis and ameliorated hypoxia.