CHU Chu;AI Shuang;ZHANG Yunhong;FAN Nannan;WANG Yurou;LI Xia;WANG Bin;Innovative Institute of Chinese Medicine and Pharmacy,Shandong University of Traditional Chinese Medicine;Affiliated Hospital of Shandong University of Traditional Chinese Medicine;Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine;Objective:To explore the mechanism of Xiaoshuan Tongmai Decoction(消栓通脉汤) regulating the janus kinase 1/signal transducer and activator of transcription 1(JAK1/STAT1) signaling pathway to alleviate dendritic cell(DC) pyroptosis in treating deep vein thrombosis(DVT). Methods:A total of 20 C57 BL/6J mice were selected to establish a mouse DVT model using inferior vena cava(IVC) stenosis method. They were then randomly divided into a model group,a low-dose group of Xiaoshuan Tongmai Decoction,a high-dose group of Xiaoshuan Tongmai Decoction,and a western medicine group,with 5 mice in each group. Each drug-treated group was given corresponding drug intervention by gavage and subcutaneous injection,while the model group received intervention of an equal amount of double distilled water by gavage for one week. Hematoxylin eosin(HE) staining and small animal Doppler color ultrasound were used to observe the size of thrombus formation in mice. The protein expressions of cleaved cysteine aspartate protease 1 p20(Caspase-1 p20) and gasdermin D N-terminal fragment(GSDMD-N) in peripheral blood mononuclear cells(PBMCs) of mice were detected by Western blot. Enzyme linked immunosorbent assay(ELISA) was used to detect the expression of serum interleukin-1β(IL-1β) and interleukin-18(IL-18). A total of 10 C57 BL/6J mice were selected to prepare Xiaoshuan Tongmai Decoction-containing serum for cell experiments. Lipopolysaccharide(LPS) and adenosine triphosphate(ATP) were used to induce pyroptosis of dendritic cells(DC2.4) in mice,and the DC2.4 cells were divided into dimethyl sulfoxide(DMSO) group,LPS+ATP+blank serum group,and LPS+ATP+high-dose Xiaoshuan Tongmai Decoction-containing serum group. Western blot was used to detect the protein expression of phosphorylated(p)-JAK1,p-STAT1,Caspase-1 p20,and GSDMD-N in DC2.4,and ELISA was used to detect the protein expression of IL-1β and IL-18 in the cell supernatant. Another 15 C57 BL/6J mice were randomly divided into a sham-operation group,a model group,and a high-dose Xiaoshuan Tongmai Decoction group,with 5 mice in each group. After gavage intervention,blood was collected,and Western blot was used to detect the protein expression of p-JAK1,p-STAT1,Caspase-1 p20,and GSDMD-N in the PBMCs of mice. Another 15 mice were randomly divided into 3 groups(5 mice in each group),namely a model group,a high-dose Xiaoshuan Tongmai Decoction+Vector group,and a high-dose Xiaoshuan Tongmai Decoction+pcDNA3.1-mJAK1-3×Flag-C group. The length and mass of thrombus in mice were measured. Results:Compared with the model group,the thrombus formation in the low-dose group of Xiaoshuan Tongmai Decoction,high-dose group of Xiaoshuan Tongmai Decoction,and western medicine group was significantly reduced,and the expression levels of Caspase-1 p20,GSDMD-N,IL-1β,and IL-18 decreased(P<0.01). Compared with the LPS+ATP+blank serum group,the protein expression levels of p-JAK1,p-STAT1,Caspase-1 p20,GSDMD-N,IL-1β,and IL-18 in the LPS+ATP+high-dose Xiaoshuan Tongmai Decoction-containing serum group decreased(P<0.05 or P<0.01). Compared with the model group,the protein expression of p-JAK1,p-STAT1,Caspase-1 p20,and GSDMD-N in the high-dose Xiaoshuan Tongmai Decoction group decreased(P<0.01). Compared with the model group,the length and mass of thrombus in the high-dose Xiaoshuan Tongmai Decoction+Vector group decreased(P<0.01). Compared with the high-dose Xiaoshuan Tongmai Decoction+Vector group,the length and mass of thrombus in the high-dose Xiaoshuan Tongmai Decoction+pcDNA3.1-mJAK1-3×Flag-C group increased(P<0.01). Conclusion:Xiaoshuan Tongmai Decoction treats DVT by inhibiting the JAK1/STAT1 signaling pathway,thereby reducing DC pyroptosis.
2025 06 v.49;No.277 [Abstract][OnlineView][Download 2247K]