Huoxue Qushi medicine mud attenuates the deep venous thrombosis formation through inhibiting the NLRP3 inflammasome-mediated inflammatory responses in platelets.

BACKGROUND

Traditional Chinese medicine (TCM) shows significant therapeutic effects in deep venous thrombosis (DVT). This study investigated the therapeutic effects and mechanism of Huoxue Qushi (HXQS) on DVT, focusing on the nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) activation.

METHODS

High performance liquid chromatography (HPLC) was used to detect the active components of HXQS medicine mud. A DVT mouse model was constructed by inferior vena cava ligation. HXQS medicine mud was applied to acupoints (Zusanli, Sanyinjiao, and Taichong), with Heparin (UFH) as a positive control. An NLRP3 overexpression (oe-NLRP3) lentivirus was performed to evaluate the specific mechanism-related to NLRP3 activation of HXQS on DVT. Hematoxylin-Eosin (HE) staining was performed to observe the venous vascular tissues. A PAP8 aggregation instrument was used to evaluate the platelet aggregation level. Flow cytometry was applied to detect the percentage of CD41+ platelet microparticles, platelet-neutrophil aggregation, and P-selectin. ELISA was adopted to detect the levels of inflammatory cytokines (IL-1β, IL-6, TNF-α, and IL-10). Western blotting was used to detect the protein levels of NLRP3 inflammasome-related proteins (NLRP3, caspase-1, IL-1β, and IL-18) in platelets.

RESULTS

HPLC indicated the existence of anti-thrombotic components (chlorogenic acid, amygdalin, and safflower yellow) in HXQS medicine mud. HXQS decreased thrombosis formation, plasma microparticles, platelet aggregation, and P-selectin in DVT mice. HXQS treatment also decreased NLRP3 inflammasome-related proteins (NLRP3, caspase-1, IL-1β, and IL-18) and inflammatory cytokines (IL-1β and TNF-α). oe-NLRP3 lentivirus injection blocked these alterations.

CONCLUSION

HXQS prescription exhibits anti-thrombotic efficacy in DVT through inhibiting NLRP3 inflammasome-driven inflammatory response, providing novel insight for DVT prevention and intervention.