益气活血方介导巨噬细胞极化影响慢性难愈性创面的机制研究  

Research on the mechanism of Yiqi Huoxue formula mediating macrophage polarization to affect chronic refractory wounds

作　　者：任悦怡 陈鑫球 张芝桐 姜自伟[2] 董航[2] 黄枫[2] REN Yueyi;CHEN Xinqiu;ZHANG Zhitong;JIANG Ziwei;DONG Hang;HUANG Feng(The First School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一临床医学院,广东广州510405 [2]广州中医药大学第一附属医院,广东广州510405

出　　处：《海南医学院学报》2024年第18期1392-1402,共11页Journal of Hainan Medical University

基　　金：全国名老中医药专家传承工作室建设项目[国中医药人教函(2022)75号];国家自然科学基金资助项目(82004390);广州中医药大学“双一流”与高水平大学学科后备人才培育项目。

摘　　要：目的:基于网络药理学、分子对接及细胞实验探究黄枫教授经验方益气活血方(YQHXF)介导巨噬细胞极化治疗慢性难愈性创面(CRW)的作用机制。方法:结合网络药理学和分子对接技术,分析并验证YQHXF影响CRW潜在靶点。CCK8检测YQHXF和LPS对Raw264.7巨噬细胞和增殖活性的影响,利用LPS建立RAW264.7细胞炎症模型,并加入JAK2信号通路激活剂香豆霉素a1(C-a1)和YQHXF含药血清干预,分组如下:Raw264.7组(对照组)、Raw264.7+LPS组(模型组)、Raw264.7+LPS+YQHXF组、Raw264.7+LPS+YQHXF+C-a1组、Raw264.7+LPS+C-a1组,流式细胞术检测Raw264.7细胞极化情况;RT-qPCR和ELISA检测炎症因子IL-4、IL-6、IL-13、TNF-α的转录及表达,Western Blot检测JAK2/STAT3通路磷酸化情况。结果:网络药理学与分子对接研究发现,YQHXF可能通过介导JAK2/STAT3信号通路调控巨噬细胞极化,调节炎症因子表达,促进CRW的愈合。细胞实验结果表明,YQHXF可显著提高巨噬细胞增殖活性(P<0.05);模型组和C-a1组JAK2和STAT3磷酸化、M1型巨噬细胞比例、IL-6和TNF-α转录和蛋白表达明显高于对照组(P<0.05);与模型组和C-a1组比较,加入YQHXF处理可显著降低JAK2和STAT3磷酸化、M1型巨噬细胞比例、IL-6和TNF-α转录和蛋白表达水平(P<0.05),提高M2型巨噬细胞比例、IL-4、IL-13转录和蛋白表达水平(P<0.05)。结论:YQHXF可能通过增强巨噬细胞增殖活性,抑制JAK2、STAT3磷酸化,正向调控巨噬细胞M2抗炎表型极化,抑制M1促炎表型极化,减轻炎症反应,达到对CRW的治疗效果。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,molecular docking,and cell experiments.Methods:Combining network pharmacology and molecular docking technology,we analyzed and verified the potential targets of YQHXF on CRW.CCK8 was used to detect the effects of YQHXF and LPS on the macrophage 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-αtranscription and expression.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-αtranscription 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 significantly reduced JAK2 and STAT3 phosphorylation,M1 macrophage ratio,IL-6,and TNF-αtranscription and protein expression levels(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 J

关 键 词：名医经验 慢性难愈性创面 网络药理学 分子对接 JAK2-STAT3 巨噬细胞极化 

分 类 号：R285.5[医药卫生—中药学]