Study on the Mechanism of Huoxue Jiedu Decoction in the Intervention of Phosphatidylinositol 3-kinase/Serine-threonine kinase/Nuclear factor-kappa B Pathway in the treatment of Deep Vein Thrombosis

Ethnopharmacology-associated: Huoxue Jiedu Decoction (HXJDT), a traditional Chinese medicine (TCM) formula derived from the TCM theory of 'blood stasis and toxin accumulation', holds significant ethnopharmacological relevance in the treating deep vein thrombosis (DVT). Our animal studies demonstrated that HXJDT regulates the lncRNA GAS5/miR-21/PTEN axis to suppress the PI3K/Akt/NF-κB inflammatory signaling pathway, thereby bridging traditional therapeutic knowledge ('Huoxue Jiedu' therapy) with modern mechanistic insights. These findings highlight the unique value of ethnomedicine in developing antithrombotic strategies with reduced bleeding risk. This study aimed to investigate the therapeutic effects and underlying molecular mechanisms of HXJDT in the treatment of DVT. A DVT model was established via the inferior vena cava stenosis method to evaluate the therapeutic effect of HXJDT. Vascular morphology following modeling was examined by hematoxylin-eosin (HE) staining, and the expression differences of M1 and M2 macrophages during thrombus formation were explored by immunohistochemistry. The expression levels of pathway-related proteins and genes were measured using ELISA kits, Western blotting, and quantitative real-time PCR (qPCR). This study revealed that the PI3K/Akt/NF-κB signaling pathway was highly expressed in the DVT model. It promoted macrophage recruitment, activated the expression of downstream pro - inflammatory factors (TNF-α, IL-6), upregulated the expression of PAI-1, and downregulated the expression of IL-10 and t-PA, thereby inhibiting thrombus dissolution. PTEN, a negative regulator of the PI3K/Akt/NF-κB pathway, exhibited decreased expression in DVT. Meanwhile, the lncRNA GAS5/miRNA-21 sponge mechanism also played an important role in DVT progression. HXJDT significantly ameliorated the pathogenesis processes of DVT. Notably, the therapeutic effect of high-concentration HXJDT was superior than that of rivaroxaban tablets. In conclusion, HXJDT regulates the lncRNA GAS5/miRNA-21/PTEN axis to suppress the PI3K/Akt/NF-κB pathway, thereby ameliorating DVT and improving prognosis, which highlights its potential clinical value.