The Dual Anti-inflammatory and Anticoagulant Effects of Jianpi Huashi Tongluo Prescription on Rheumatoid Arthritis through Inhibiting the Activation of the PI3K/AKT Signaling Pathway

Fanfan Wang Jian Liu ^*

• Anhui University of Chinese Medicine, Hefei, China

Background: Rheumatoid arthritis (RA) is often accompanied by abnormal changes in inflammatory responses and coagulation-fibrinolysis indicators. Jianpi Huashi Tongluo Prescription -Xinfeng Capsule (XFC) is widely used clinically for the treatment of RA. It exhibits dual anti-inflammatory and anticoagulant effects.However, the specific mechanisms underlying its actions remain to be further investigated.Objective: This study aims to elucidate the anti-inflammatory and anticoagulant mechanisms of XFC in the treatment of RA.Methods: A multidimensional methodological framework was employed. Firstly, through retrospective clinical data mining, combined with the Apriori algorithm and random walk models, an in-depth analysis was conducted to explore the potential associations between XFC treatment and improvements in clinical inflammatory and coagulation markers among RA patients. Secondly, an adjuvant-induced arthritis rat model was established to directly observe the anti-inflammatory and anticoagulant effects of XFC in vivo. Lastly, a co-culture system of RA patient-derived peripheral blood mononuclear cells (RA-PBMCs) and vascular endothelial cells (VECs) was established to mimic the in vivo microenvironment, and the anti-inflammatory and anticoagulant mechanisms of XFC were validated in vitro.Results: Data mining analysis revealed abnormally elevated levels of inflammatory and coagulation markers such as fibrinogen (FBG), erythrocyte sedimentation rate (ESR), high-sensitivity C-reactive protein (Hs-CRP), and rheumatoid factor (RF) in RA patients (p<0.001), and emphasized the close correlation between XFC treatment and the improvement of these markers including Hs-CRP, ESR, and RF (confidence >60% and lift >1). Animal experimental data indicated that XFC effectively reduced the levels of inflammatory and coagulant markers (IL-6, D-D, FBG, PAF, VEGF, and TF) in adjuvant-induced arthritis (AA) rats while enhancing the expression of anti-inflammatory factors (IL-10) (p<0.05). Additionally, network pharmacology and molecular docking results show that the main active components of XFC, namely calycosin-7-O-beta-D-glucoside, calycosin, and formononetin, exhibit excellent docking with the core targets HIF1A, PTGS2, and MMP9. In vitro co-culture model showed that XFC inhibited RA-related inflammatory responses and hypercoagulable states by suppressing the activation of the PI3K/AKT signaling pathway.This study demonstrates that XFC exerts its dual anti-inflammatory and anticoagulant effects, at least in part, by inhibiting the activation of the PI3K/AKT signaling pathway, providing potential insights into targeted therapy for RA.