摘要
目的:探究紫锥菊多糖对克雷伯菌诱导的肺炎模型大鼠肺部炎症损伤的影响,并探究其抗炎机制。方法:用克雷伯菌制备肺炎大鼠模型,分为模型对照组、紫锥菊多糖5、10 mg/kg组、Toll样受体4(RS09)激活剂25μg/kg组、紫锥菊多糖10 mg/kg+TLR4激活剂25μg/kg组,每组20只。各组大鼠给予相应药物干预14 d。动物肺功能分析仪检测大鼠肺功能指标(肺活量、肺阻力、肺顺应性);ELISA法检测大鼠肺泡灌洗液中TLR4、T细胞免疫球蛋白黏蛋白分子-3(TIM-3)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)含量;传统培养法测定肺泡灌洗液中细菌含量;全自动血细胞分析仪和显微镜测定或观察肺泡灌洗液中白细胞(WBC)和多形核中性粒细胞(PMN)数量;HE染色观察大鼠肺组织病理损伤;免疫组化法检测大鼠肺组织M1型巨噬细胞极化标记蛋白(CD11)阳性表达;Western blot法检测大鼠肺组织CD11、M2型巨噬细胞极化标记蛋白(CD206)、T盒子转录因子(T-BET)、GATA结合蛋白3(GATA3)、细菌菌毛黏附蛋白(FIMH)、TLR4、磷酸化-NF-κB(p-NF-κB)、核因子-κB(NF-κB)、TIM-3蛋白表达。结果:与正常对照组比较,模型对照组大鼠可见肺泡壁充血、支气管周围炎性细胞浸润明显,病变严重区域连成片,肺活量、肺顺应性显著降低,肺组织CD206、GATA3蛋白表达显著下调,肺阻力、肺泡灌洗液中TLR4、TIM-3、TNF-α、IL-6含量、细菌数量、WBC、PMN数量、肺组织CD11阳性表达显著升高,肺组织CD11、T-BET、FIMH、TLR4、p-NF-κB/NF-κB、TIM-3蛋白表达显著上调(P<0.01);与模型对照组比较,紫锥菊多糖5、10 mg/kg组大鼠肺泡损伤及炎性细胞浸润等病理损伤均呈不同程度缓解,肺活量、肺顺应性升高,肺组织CD206、GATA3蛋白表达上调,肺阻力、肺泡灌洗液中TLR4、TIM-3、TNF-α、IL-6含量、细菌数量、WBC、PMN数量、肺组织CD11阳性表达下调,肺组织CD11、T-BET、FIMH、TLR4、p-NF-κB/NF-κB、TIM-3蛋白表达下调(P<0.01);RS0925μg/kg组大鼠肺泡破裂,炎性细胞浸润最多,病变严重区域连成片,肺活量、肺顺应性、肺组织CD206、GATA3蛋白表达显著下调,肺阻力、肺泡灌洗液中TLR4、TIM-3、TNF-α、IL-6含量、细菌数量、WBC、PMN数量、肺组织CD11阳性表达升高,肺组织CD11、T-BET、FIMH、TLR4、p-NF-κB/NF-κB、TIM-3蛋白表达显著上调(P<0.01);紫锥菊多糖10 mg/kg+RS0925μg/kg组大鼠肺泡损伤及炎性细胞浸润等病理变化和上述定量指标较紫锥菊多糖10 mg/kg组恶化,较RS0925μg/kg组显著改善(P<0.01)。结论:紫锥菊多糖可能通过抑制TLR4/NF-κB通路,改善肺炎克雷伯菌诱导的肺炎模型大鼠肺功能,提高抗炎免疫细胞活化,缓解肺组织炎症及病理损伤。
Objective:To investigate the effect of echinacea polysaccharide on pulmonary inflammation injury in a Klebsiella pneumoniae-induced pneumonia rat model and explore its anti-inflammatory mechanism.Methods:A pneumonia rat model was established using Klebsiella pneumoniae and divided into five groups:model control group,echinacea polysaccharide 5 mg/kg group,echinacea polysaccharide 10 mg/kg group,Toll-like receptor 4(TLR4)activator 25μg/kg group,and echinacea polysaccharide 10 mg/kg+TLR4 activator 25μg/kg group,with 20 rats in each group.Each group received corresponding interventions for 14 days.Lung function parameters(lung capacity,lung resistance,and lung compliance)were detected by an animal lung function analyzer.The levels of TLR4,T cell immunoglobulin mucin molecule-3(TIM-3),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in bronchoalveolar lavage fluid(BALF)were detected by enzyme-linked immunosorbent assay(ELISA).Bacterial counts in BALF were determined by traditional culture methods.White blood cell(WBC)and polymorphonuclear neutrophil(PMN)counts in BALF were determined by an automated hematology analyzer and a microscope,respectively.Pathological injury of lung tissue was observed by hematoxylin-eosin staining.Immunohistochemistry was used to detect CD11-positive expression,a marker of M1 macrophage polarization.Western blotting was employed to detect the expression of CD11,CD206(M2 macrophage marker),T-box transcription factor(T-BET),GATA binding protein 3(GATA3),fimbrial adhesion protein(FIMH),TLR4,phosphorylated NF-κB(p-NF-κB),total NF-κB,and TIM-3 in lung tissue.Results:Compared with the normal control group,the model control group showed alveolar wall congestion,significant infiltration of inflammatory cells around the bronchi,and extensive areas of lesions.Lung capacity,lung compliance,and the expression of CD206 and GATA3 in lung tissue were significantly decreased,while lung resistance,levels of TLR4,TIM-3,TNF-α,IL-6,bacterial count,WBC,and PMN in BALF,and the expression of CD11-positive cells,CD11,T-BET,FIMH,TLR4,p-NF-κB/NF-κB,and TIM-3 proteins were significantly increased(P<0.01).Compared with the model control group,pathological damage,including alveolar injury and inflammatory infiltration in the echinacea polysaccharide 5 mg/kg and 10 mg/kg groups was alleviated to varying degrees.Lung capacity,lung compliance,and the expression of CD206 and GATA3 in lung tissue were increased in a dose-dependent manner,while lung resistance,BALF levels of TLR4,TIM-3,TNF-α,IL-6,bacterial count,WBC,PMN,and expression of CD11-positive cells,CD11,T-BET,FIMH,TLR4,p-NF-κB/NF-κB,and TIM-3 were decreased successively(P<0.01).Rats in the TLR4 activator group exhibited severe alveolar rupture,the most extensive inflammatory infiltration,and the largest areas of lesions,with significantly decreased lung capacity,compliance,CD206,and GATA3 expression,and significantly increased indicators of inflammation(P<0.01).In the echinacea polysaccharide 10 mg/kg+TLR4 activator 25μg/kg group,pathological changes and inflammatory markers worsened compared to the echinacea polysaccharide 10 mg/kg group but were improved relative to the TLR4 activator 25μg/kg group alone(P<0.01).Conclusions:Echinacea polysaccharide may improve lung function in the Klebsiella pneumoniae-induced pneumonia model rats by inhibiting the TLR4/NF-κB pathway,enhancing anti-inflammatory immune cell activation,and relieving lung inflammation and pathological damage.
作者
高凡凡
马猛
李妍
王宁
GAO Fanfan;MA Meng;LI Yan;WANG Ning(Tianjin Medical University Chu Hsien-I Memorial Hospital,Tianjin Institute of Endocrinology,National Health Commission Key Laboratory of Hormones and Development,Tianjin Key Laboratory of Metabolic Diseases,Tianjin 300134)
出处
《中药药理与临床》
北大核心
2026年第3期89-96,共8页
Pharmacology and Clinics of Chinese Materia Medica
基金
天津市自然科学基金项目(编号:23JCYBJC00134)。
关键词
紫锥菊多糖
肺炎
肺功能
炎症损伤
TOLL样受体4
核因子-ΚB
Echinacea polysaccharide
Pneumonia
Lung function
Inflammatory injury
Toll-like receptor 4
Nuclear factor-κB
