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

作　　者：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[1] YANG Hongjuan;YANG Yaru;YANG Yujie;ZHU Zhongbo;MA Quan;WU Yanlin;LI Hongmei;ZHANG Xuhui;LIU Xiping(Gansu Provincial Engineering Laboratory for the Creation of New Traditional Chinese Medicine(TCM)Products,Key Laboratory of Excavation and Innovative Transformation of TCM in Gansu Province,Gansu University of Chinese Medicine,Lanzhou 730000,China;Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[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

出　　处：《中国实验方剂学杂志》2025年第1期146-155,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：甘肃省科技重大项目(22ZD1FA001);国家自然科学基金项目(82260889);第五批全国中医临床优秀人才研修项目(国中医药人教函【2022】239号);甘肃省自然科学基金项目(24JRRD002)。

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,14 d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、IFObjective: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 STAT1,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 level

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 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R256