中国实验方剂学杂志2025，Vol.31Issue(1) ：146-155.DOI:10.13422/j.cnki.syfjx.20241405

基于STAT1/IRF3信号通路探讨益肺健脾方对脂多糖诱导的大鼠肺部免疫炎症反应的影响

Effect of Yifei Jianpi Prescription on Lipopolysaccharide-induced Lung Immune Inflammatory Response in Rats Based on STAT1/IRF3 Pathway

杨红娟 杨亚茹 杨玉洁 朱中博 马泉 武妍琳 李红梅 张旭辉 刘喜平
中国实验方剂学杂志2025，Vol.31Issue(1) ：146-155.DOI:10.13422/j.cnki.syfjx.20241405
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基于STAT1/IRF3信号通路探讨益肺健脾方对脂多糖诱导的大鼠肺部免疫炎症反应的影响

Effect of Yifei Jianpi Prescription on Lipopolysaccharide-induced Lung Immune Inflammatory Response in Rats Based on STAT1/IRF3 Pathway

杨红娟 1杨亚茹 1杨玉洁 1朱中博 1马泉 2武妍琳 2李红梅 2张旭辉 2刘喜平1
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作者信息

  • 1. 甘肃中医药大学甘肃省中药新产品创制工程实验室,甘肃省中医方药挖掘与创新转化重点实验室,兰州 730000
  • 2. 甘肃中医药大学附属医院,兰州 730000
  • 折叠

摘要

目的:观察益肺健脾方对脂多糖(LPS)诱导的肺炎模型大鼠信号传导转录激活因子1(STAT1)/干扰素调节因子3(IRF3)信号通路的影响,探讨益肺健脾方改善肺部免疫炎症反应的机制.方法:将60只雄性SPF级SD大鼠,随机抽取10只为正常组,余50只气管滴注脂多糖建立大鼠肺炎模型,成功后再随机分为模型组、地塞米松组(0.5 mg·kg-1)、益肺健脾方高、中、低(12、6、3 mg·kg-1)剂量组,每组10只,每天给药1次,正常组和模型组予等体积生理盐水,14d后,流式细胞术检测全血淋巴细胞分类,酶联免疫吸附测定法(ELISA)测定血清免疫球蛋白G(IgG)、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)及肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)含量,苏木素-伊红(HE)染色观察大鼠肺组织病理改变并评分,计算脾脏、胸腺重量及肺湿干重比(W/D),实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织信号STAT1、IRF3、IL-6、干扰素-α(IFN-α)mRNA表达,蛋白免疫印迹法(Western blot)检测肺组织STAT1、IRF3、IL-6、IFN-α蛋白表达.结果:与正常组比较,模型组大鼠外周血B淋巴细胞比例显著升高,NK细胞比例、CD4+/CD8+降低(P＜0.05,P＜0.01),血清IgG、IgA含量降低,IgM含量显著升高(P＜0.01),BALF中TNF-α、IL-6、IL-8含量显著升高,IL-10含量显著降低(P＜0.01),肺组织损伤明显,胸腺、脾脏质量显著增加,W/D值升高(P＜0.01),肺组织STAT1、IRF3、IFN-α、IL-6 mRNA及蛋白表达明显上调(P＜0.05,P＜0.01).与模型组比较,益肺健脾方各组大鼠外周血B淋巴细胞比例显著降低,NK细胞比例、CD4+/CD8+升高(P＜0.05,P＜0.01),血清IgG、IgA含量明显升高,IgM含量明显下降(P＜0.05,P＜0.01),BALF中TNF-α、IL-6、IL-8表达显著降低,IL-10表达显著升高(P＜0.01),肺组织损伤程度减轻,胸腺、脾脏质量显著减轻,W/D值显著降低(P＜0.01),肺组织STAT1、IRF3、IFN-α、IL-6mRNA及蛋白表达明显下调(P＜0.05,P＜0.01).结论:益肺健脾方可减轻脂多糖诱导的肺炎大鼠肺组织损伤,改善肺部免疫炎症反应,其机制可能与抑制STAT1/IRF3信号通路激活有关.

Abstract

Objective:To observe the effect of Yifei Jianpi prescription on the of signal transducer and activator of transcription protein 1(STAT1)/interferon regulatory factor 3(IRF3)signaling pathway in a pneumonia model induced by lipopolysaccharide(LPS)and to explore the mechanism of Yifei Jianpi prescription in improving lung immune and inflammatory responses.Methods:Sixty male SPF SD rats were used in this study.Ten rats were randomly assigned to the normal control group,and the remaining 50 were instilled with LPS in the trachea to establish a pneumonia model.After successful modeling,the rats were randomly divided into the model group,dexamethasone group(0.5 mg·kg-1),and Yifei Jianpi prescription high-dose(12 mg·kg-1),medium-dose(6 mg·kg-1),and low-dose(3 mg·kg-1)groups,with 10 rats in each group.Treatment was administered once daily,and the normal control and model groups received the same volume of normal saline.After 14 days,flow cytometry was used to detect the classification of whole blood lymphocytes.Enzyme-linked immunosorbent assay(ELISA)was used to measure serum levels of immunoglobulin G(IgG),immunoglobulin A(IgA),immunoglobulin M(IgM),and the content of tumor necrosis factor-α(TNF-α),interleukin-8(IL-8),interleukin-6(IL-6),and interleukin-10(IL-10)in alveolar lavage fluid(BALF).Hematoxylin-eosin(HE)staining was used to observe lung tissue pathology and score the damage.Thymus weight,spleen weight,and wet-to-dry weight ratio(W/D)were recorded.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of STAT1,IRF3,IL-6,and interferon-alpha(IFN-α)in lung tissues,while Western blot was performed to assess the protein expression of STAT 1,IRF3,IL-6,and IFN-α.Results:Compared with the normal control group,the model group showed significantly increased proportion of B lymphocytes in peripheral blood,decreased proportions of NK cells and CD4+/CD8+(P＜0.05,P＜0.01),decreased serum levels of IgG and IgA,significantly increased IgM levels(P＜0.01),significantly elevated content of TNF-α,IL-6,and IL-8 in BALF,and significantly decreased IL-10 levels(P＜0.01).Lung tissue damage was evident,with significant increases in thymus and spleen weights and a higher W/D ratio(P＜0.01).The mRNA and protein expression of STAT1,IRF3,IFN-α,and IL-6 in lung tissues was significantly upregulated(P＜0.05,P＜0.01).Compared with the model group,the Yifei Jianpi prescription groups showed significantly reduced proportions of B lymphocytes in peripheral blood,increased proportions of NK cells and CD4+/CD8+ratios(P＜0.05,P＜0.01),significantly increased serum levels of IgG and IgA,significantly decreased IgM levels(P＜0.05,P＜0.01),significantly reduced levels of TNF-α,IL-6,and IL-8 in BALF,and significantly increased IL-10 levels(P＜0.01).Lung tissue damage was alleviated,thymus and spleen weights were significantly reduced,and the W/D ratio was markedly decreased(P＜0.01).The mRNA and protein expression of STAT1,IRF3,IFN-α,and IL-6 in lung tissues was significantly downregulated(P＜0.05,P＜0.01).Conclusion:Yifei Jianpi prescription can alleviate lung tissue damage and improve immune and inflammatory responses in LPS-induced pneumonia rats.The mechanism may be related to the inhibition of STAT1/IRF3 signaling pathway activation.

关键词

益肺健脾方/急性肺损伤/免疫炎症反应/信号传导转录激活因子1/干扰素调节因子3(STAT1/IRF3)信号通路

Key words

Yifei Jianpi prescription/acute lung injury/immune inflammatory response/signal transducer and activator of transcription protein 1/interferon regulatory factor 3(STAT1/IRF3)signaling pathway

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出版年

2025
中国实验方剂学杂志
中国中医科学院中药研究所　中国中西医结合学会中药专业委员会

中国实验方剂学杂志

CSCD北大核心
影响因子：1.62
ISSN：1005-9903
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