河南中医药大学
河南省自然基金面上项目(232300420054)；河南省科技研发计划联合基金(No.222301420088)；河南省高校重点科研项目(No.23A360013)
Henan University of Chinese Medicine
【】目的 探讨茯苓杏仁甘草汤(Fuling Xingren Gancao granule,FXG)对小鼠动脉粥样硬化(atherosclerosis,AS)巨噬细胞极化的调控作用。方法 ApoE-/-小鼠构建AS模型,RAW264.7巨噬细胞构建极化模型。油红O和苏木精-伊红染色法 ( hematoxylin-eosin staining,HE )染色观察主动脉斑块总面积和主动脉管腔狭窄程度。PCR和Western blot体内外检测M1型极化因子诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、单核细胞趋化蛋白1(monocyte chemotactic protein 1, CCL2)和M2型极化因子精氨酸酶-1(arginase-1,Arg-1)、类几丁质酶样蛋白(Chitinase-Like Protein 3,YM1)、甘露糖受体(macrophage mannose receptor1,Mrcl或称CD206)的表达水平及信号转导和转录激活因子3(signal transducer and activator of transcription,STAT3)磷酸化水平。结果 FXG明显减少ApoE-/-小鼠主动脉斑块总面积、降低M1型巨噬细胞极化因子iNOS、CCL2表达水平,升高M2型巨噬细胞极化因子Arg-1、YM1表达水平(P<0.05)。模型组小鼠和M1型巨噬细胞STAT3磷酸化水平降低,在FXG干预后其磷酸化水平上调(P<0.05)。P-STAT3抑制剂Stattic部分消除了FXG对iNOS和Arg-1的调控作用(P<0.05)。结论 FXG具有抑制AS进展的作用,其作用与靶向STAT3调控巨噬细胞极化有关。
Objective To explore the regulatory effect of Fuling Xingren Gancao granule (FXG) on macrophage polarization in atherosclerosis (AS) mice. Methods ApoE -/- mice were used to construct an AS model, while RAW264.7 macrophages were used to construct a polarized model. Oil red O and hematoxylin eosin staining (HE) were used to observe the total area of aortic plaques and the degree of aortic stenosis. PCR and Western blot were used to detect the expression levels, signal transduction, and transcription of M1 polarization factor inducible nitric oxide synthase (iNOS), monocyte chemotactic protein 1 (CCL2), M2 polarization factor arginase-1 (Arg-1), chitinase like protein 3 (YM1), and mannose receptor 1 (Mrcl or CD206) in vitro and in vivo. The phosphorylation level of signal transducer and activator of transcription (STAT3). Result: FXG significantly reduced the total area of aortic plaques in ApoE -/- mice, decreased the expression levels of M1 macrophage polarization factors iNOS and CCL2, and increased the expression levels of M2 macrophage polarization factors Arg-1 and YM1 (P<0.05). The phosphorylation levels of STAT3 in the model group mice and M1 macrophages decreased, but were upregulated after FXG intervention (P<0.05). The P-STAT3 inhibitor Stattic partially eliminated the regulatory effect of FXG on iNOS and Arg-1 (P<0.05).Result FXG significantly reduced the total area of aortic plaques in ApoE -/- mice, decreased the expression levels of M1 macrophage polarization factors iNOS and CCL2, and increased the expression levels of M2 macrophage polarization factors Arg-1 and YM1 (P<0.05). The phosphorylation levels of STAT3 in the model group mice and M1 macrophages decreased, but were upregulated after FXG intervention (P<0.05). The P-STAT3 inhibitor Stattic partially eliminated the regulatory effect of FXG on iNOS and Arg-1 (P<0.05).Conclusion FXG has an inhibitory effect on the progression of AS, which is related to the targeted regulation of macrophage polarization by STAT3.
