Research on the mechanism of Yiqi Huoxue formula mediating macrophage polarization to affect chronic refractory wounds
Objective:To explore the mechanism of Professor Huang Feng's empirical formula Yiqi Huoxue Formula(YQHXF)mediated macrophage polarization therapy for chronic refractory wounds(CRW)based on network pharmacology,mo-lecular docking,and cell experiments.Methods:Combining network pharmacology and molecular docking technology,we ana-lyzed and verified the potential targets of YQHXF on CRW.CCK8 was used to detect the effects of YQHXF and LPS on the mac-rophage and proliferation activity of Raw264.7 cells.A RAW264.7 cell inflammation model was established using LPS,and JAK2 signaling pathway activator coumarin a1(C-a1)and YQHXF drug containing serum intervention were added.The groups were as follows:Raw264.7 group(control group),Raw264.7+LPS group(model group),Raw264.7+LPS+YQHXF group,Raw264.7+LPS+YQHXF+C-a1 group,and Raw264.7+LPS+C-a1 group.Flow cytometry was used to detect the polarization of Raw264.7 cells;RT qPCR and ELISA detected of inflammatory factors IL-4,IL-6,IL-13,TNF-α transcription and expres-sion.Western blot was used to detect the JAK2/STAT3 pathway phosphorylation.Results:Network pharmacology and molecular docking studies found that YQHXF may regulate macrophage polarization,regulate the expression of inflammatory factors,and promote the healing of CRW by mediating the JAK2/STAT3 signaling pathway.The results of cell experiments showed that YQHXF can significantly enhance macrophage proliferation activity(P<0.05);In model group and C-a1 group,JAK2 and STAT3 phosphorylation,M1 macrophage ratio,IL-6 and TNF-α transcription and protein expression were significantly higher than the control group(P<0.05);Compared with the model group and C-a1 group,the addition of YQHXF treatment significant-ly reduced JAK2 and STAT3 phosphorylation,M1 macrophage ratio,IL-6,and TNF-α transcription and protein expression lev-els(P<0.05),increased the proportion of M2 macrophages,as well as the transcription and protein expression levels of IL-4 and IL-13(P<0.05).Conclusion:YQHXF may enhance macrophage proliferation activity,inhibit JAK2 and STAT3 phosphoryla-tion,positively regulate macrophage M2 anti-inflammatory phenotype polarization,inhibit M1 pro-inflammatory phenotype polar-ization,alleviate inflammatory response,and achieve therapeutic effects.
Experience of renowned doctorsChronic refractory woundsNetwork pharmacologyMolecular dockingJAK2-STAT3Macrophage polarization
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