如意金黄散通过自噬途径调控NLRP3炎症小体表达治疗皮肤脓肿的机制研究OA
Mechanistic Investigation of Ruyi Jinhuang Powder in Regulating NLRP3 Inflammasome Via the Autophagy Pathway for the Treatment of Skin Abscess
目的:通过金黄色葡萄球菌感染建立皮肤脓肿小鼠模型,评估如意金黄散对皮肤脓肿小鼠自噬及 NOD样受体热蛋白结构域相关蛋白3(NLRP3)表达的干预作用及作用机制.方法:采用苏木素-伊红(HE)染色法观察组织病理形态;通过透射电镜观察自噬体形成;采用酶联免疫吸附测定法(ELISA)检测组织IL-18、TNF-α的含量;采用蛋白免疫印迹法(Western blot,WB)、免疫荧光染色法检测组织Beclin1、LC3Ⅱ/Ⅰ、NLRP3的蛋白表达情况.结果:HE染色结果显示,与Sham 组比较,Model组皮肤结构严重异常,表皮结构未见,真皮层暴露(红色箭头),组织内有大量炎症细胞浸润(绿色箭头),局部可见肉芽组织生成;小鼠血清中IL-18、TNF-α水平显著上调(P<0.001,P<0.001);Western blot法及免疫荧光检测法均提示Beclin1、LC3Ⅱ/Ⅰ、NLRP3水平均显著升高(P<0.05,P<0.05,P<0.001).经如意金黄散干预10d后,与 Model组 比 较,Model+RYHJS组皮肤结构改善、组织炎症细胞浸润程度较 Model组相对减弱,局部可见少量肉芽组织生成;透射电镜观察显示,如意金黄散干预后(Model+RYHJS组),小鼠皮肤组织中可见自噬小体比例升高;血清中IL-18、TNF-α水平显著下调(P<0.05,P<0.001);Western blot法及免疫荧光检测法均提示如意金黄散干预可以显著降低 NLRP3蛋白表达(P<0.001),显著提高自噬相关基因 Beclin1、LC3Ⅱ/Ⅰ表达水平(P<0.001,P<0.05).给予3-MA后,Western blot法及免疫荧光检测法均提示 NLRP3表达显著升高(P<0.01).结论:研究表明,如意金黄散可能通过自噬途径抑制 NLRP3炎症小体信号通路表达,从而治疗皮肤脓肿.
Objective:Establish a mouse model of skin abscess infected with Staphylococcus aureus and evaluate the in-tervention effect and mechanism of Ruyi Jinhuang Powder on autophagy and NOD-like receptor protein 3(NLRP3 in-flammasome)expression in mice with skin abscess.Methods:Using hematoxylin eosin(HE)staining method to ob-serve the histopathological morphology of tissues;Observation of autophagosome formation under transmission elec-tron microscopy;Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of IL-18 and TNF-α in tissues;Western blot and immunofluorescence staining were used to detect the protein expression of Beclin1,LC3 Ⅱ/Ⅰ,and NLRP3 in tissues.Results:The HE staining results showed that compared with the Sham group,the skin structure of the model group was severely abnormal,with no visible epidermal structure and exposed dermis layer(red arrow).There were a large number of inflammatory cells infiltrating the tissue(green arrow),and granulation tissue formation was observed locally;The levels of IL-18 and TNF-α in mouse serum were significantly upregulated(P<0.001,P<0.001);Western blot and immunofluorescence detection both showed significant increases in Beclin1,LC3Ⅱ/Ⅰ,and NLRP3 levels(P<0.05,P<0.05,P<0.001).After 10 days of intervention with Ruyi Jinhuang Powder,compared with the model group,the skin structure of the Model+RYHJS group improved,the degree of tissue inflam-matory cell infiltration was relatively reduced,and a small amount of granulation tissue was observed locally;Transmis-sion electron microscopy observation showed that after intervention with Ruyi Jinhuang Powder,an increase in the pro-portion of autophagosomes was observed in the skin tissue of mice;The levels of IL-18 and TNF-α in serum were sig-nificantly downregulated(P<0.05,P<0.001);Western blot and immunofluorescence detection both suggest that in-tervention with Ruyi Jinhuang Powder can significantly reduce the expression of NLRP3 protein(P<0.001)and sig-nificantly increase the expression levels of autophagy related genes Beclin1 and LC3 Ⅱ/Ⅰ(P<0.001,P<0.05).Af-ter administering 3-MA,both Western blot and immunofluorescence detection methods showed a significant increase in NLRP3 expression(P<0.01).Conclusion:Studies suggest that Ruyi Jinhuang Powder may exert therapeutic effects against skin abscesses by suppressing NLRP3 inflammasome signaling through the autophagy pathway.
王重阳;刘首记;马瑛;王姝理;谢敏;曾娟妮
湖南中医药高等专科学校附属第一医院/湖南省直中医医院,湖南 株洲 412000||湖南中医药大学,湖南 长沙 410000湖南中医药高等专科学校附属第一医院/湖南省直中医医院,湖南 株洲 412000湖南中医药高等专科学校附属第一医院/湖南省直中医医院,湖南 株洲 412000湖南中医药高等专科学校附属第一医院/湖南省直中医医院,湖南 株洲 412000湖南中医药高等专科学校附属第一医院/湖南省直中医医院,湖南 株洲 412000湖南中医药大学第二附属医院,湖南 长沙 410000
医药卫生
如意金黄散自噬NLRP3脓肿
Ruyi Jinhuang PowderautophagyNLRP3abscess
《亚太传统医药》 2026 (8)
24-31,8
湖南省教育厅科学研究项目(24C1178)
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